A traditional Chinese medicine composition for treating or / and preventing postoperative intestinal adhesion after abdominal surgery, and a preparation method and use thereof

Abstract

The application provides a traditional Chinese medicine composition for treating or / and preventing intestinal adhesion after abdominal surgery and a preparation method and application thereof, and belongs to the field of traditional Chinese medicines. The traditional Chinese medicine composition is prepared from raw medicinal materials in the following weight ratio: 8-12 parts of rhizoma sparganii, 8-12 parts of zedoary turmeric, 8-12 parts of persicae semen, 8-12 parts of safflower, 24-36 parts of radix paeoniae alba, 8-12 parts of rhizoma corydalis, 8-12 parts of fructus aurantii, 8-12 parts of olibanum, 8-12 parts of myrrh, 12-18 parts of poria cocos, 12-18 parts of atractylodes, and 4.8-7.2 parts of licorice. The application further provides a preparation method and application of the traditional Chinese medicine composition. The traditional Chinese medicine composition is used for treating or / and preventing intestinal adhesion after abdominal surgery, especially for intestinal adhesion of the type of qi stagnation and blood stasis, has a remarkable drug effect, can achieve a drug effect equivalent to that of chemical drugs such as the chemical drug prednisone acetate and itopride, and the toxicological test proves that the traditional Chinese medicine composition is safer, thereby providing a new drug selection for clinic.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery, belonging to the field of traditional Chinese medicine. Background Technology

[0002] Postoperative intestinal adhesions are a common complication after abdominal surgery, accounting for approximately 70% to 80% of all cases, especially pelvic and lower abdominal surgeries. Severe adhesions can lead to intestinal obstruction and even perforation. If postoperative patients experience abdominal pain and distension unresponsive to medical treatment, a second surgery is required to treat the obstruction, placing a significant physical and financial burden on the patient and severely impacting prognosis and surgical outcomes. Therefore, the prevention and treatment of postoperative intestinal adhesions is a pressing issue in the field of surgery.

[0003] Modern research indicates that reduced fibrin activity plays a crucial role in the development of intestinal adhesions. Normal peritoneum possesses fibrinolytic activity, composed of a series of interconnected activating and inhibiting enzymes. When peritoneal injury causes inflammation or ischemia, it produces plasma plasminogen activator inhibitors (PAIs), including PAI-1 and PAI-2. These inhibitors lead to a loss of fibrin activity in mesothelial cells. Prolonged exposure to these inhibitors can cause permanent fibrous adhesions. Therefore, intestinal adhesions are a physiological response, a type of fibrous inflammatory reaction. One of the determining factors for whether a local inflammatory response leads to fibrous adhesions is the rapid absorption of the fibrin network, which halts fibrosis and prevents adhesion formation. Conversely, fibroblasts undergo collagen degeneration, becoming the basis for fibrous adhesions and ultimately leading to intestinal adhesions. Postoperative abdominal adhesions are thus caused by a disruption of the balance between fibrinogen release and fibrinolysis in peritoneal mesothelial cells, leading to fibroblast proliferation and increased collagen secretion. Fibroblasts and newly formed capillaries grow into the tissue and, after organization, form fibrous adhesions. Various clinically used anti-adhesion products are developed to target the mechanisms of adhesion formation and inhibit different stages of adhesion production, which is also the theoretical basis of anti-adhesion drugs.

[0004] Traditional Chinese medicine (TCM) does not have a specific disease term for "postoperative intestinal adhesions." Based on its clinical manifestations, it falls under the category of "abdominal pain" in TCM. TCM possesses unique advantages and has accumulated extensive experience in the prevention and treatment of intestinal adhesions. Utilizing the multi-faceted and multi-target effects of TCM compound formulas, there is great potential for developing TCM methods to prevent and treat postoperative intestinal adhesions.

[0005] Modern treatment methods mainly include the iteration of new surgical instruments to reduce wound size, intraoperative antibiotic irrigation, intraperitoneal anticoagulation, placement of intestinal spacers, and postoperative use of anticoagulants, fibrinolytic agents, and small bowel motility-promoting drugs. However, postoperative intestinal adhesions are still difficult to avoid. Regarding chemical drugs, glucocorticoids (prednisone, dexamethasone) are classic drugs used for intestinal adhesions after abdominal surgery. Prednisone acetate's mechanism of action is potent anti-inflammatory, inhibiting fibroblast proliferation, and reducing collagen deposition. Short-term postoperative use can significantly reduce peritoneal inflammation and is one of the most widely used anti-adhesion drugs in clinical research; however, the risk of infection must be weighed, and it is usually used in short-term, low-dose applications. Dexamethasone acetate, similar to prednisone, has a stronger and longer-lasting anti-inflammatory effect and is also used to inhibit postoperative peritoneal inflammation and fibrosis, but its long half-life requires more caution when used postoperatively. The core contradiction in using glucocorticoids for the prevention and treatment of postoperative intestinal adhesions lies in the fact that while it aims to suppress inflammation and prevent adhesions, it also suppresses immunity and wound healing—and abdominal surgery is precisely where infection and non-healing wounds are most prevalent risks.

[0006] Existing traditional Chinese medicine formulas for treating intestinal adhesions mostly use herbs that promote blood circulation, remove blood stasis, regulate qi, and relieve pain. However, the formula structure is loose and lacks precise matching targeting the core pathogenesis of abdominal pain due to qi stagnation and blood stasis caused by intestinal adhesions (e.g., patent application number: 01129666.6, invention title: traditional Chinese medicine prescription for preventing and treating postoperative adhesions and method for preparing traditional Chinese medicine using it). Some formulas even contain endangered species (e.g., pangolin, which is now banned), herbs containing aristolochic acid (e.g., Aristolochia decoction, which is nephrotoxic), or laxatives (e.g., rhubarb, croton, which can easily mask the condition of intestinal obstruction) (e.g., Zhao Jinghua et al., Adhesion-relieving decoction for treating postoperative intestinal adhesions, Henan Journal of Traditional Chinese Medicine, 1996-07-20). All of these formulas pose problems with medication safety and sustainability. Moreover, the principal herbs are mostly herbs that tonify qi and nourish blood, such as angelica, ginseng, and astragalus, which are not strong enough to break down the stagnation and accumulation in the intestines and cannot meet the need for short-term clinical efficacy for abdominal pain caused by intestinal adhesions after abdominal surgery. Summary of the Invention

[0007] This invention provides a traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery, as well as its preparation method and uses.

[0008] This invention provides a traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery, which is prepared from the following raw materials in the indicated weight ratios:

[0009] 8-12 parts of Sparganium rhizome, 8-12 parts of Curcuma rhizome, 8-12 parts of Peach kernel, 8-12 parts of Safflower, 24-36 parts of White peony root, 8-12 parts of Corydalis rhizome, 8-12 parts of Sichuan Chinaberry, 8-12 parts of Frankincense, 8-12 parts of Myrrh, 12-18 parts of Poria cocos, 12-18 parts of Atractylodes macrocephala, and 4.8-7.2 parts of Licorice root.

[0010] Preferably, it is prepared from the following raw materials in the indicated weight ratios:

[0011] 10 parts of Sparganium rhizome, 10 parts of Curcuma rhizome, 10 parts of Peach kernel, 10 parts of Carthamus tinctorius, 30 parts of Paeonia lactiflora, 10 parts of Corydalis yanhusuo, 10 parts of Melia toosendan, 10 parts of Boswellia carterii, 10 parts of Commiphora myrrha, 15 parts of Poria cocos, 15 parts of Atractylodes macrocephala, and 6 parts of Glycyrrhiza uralensis.

[0012] Among them, the Corydalis Rhizome is vinegar-processed Corydalis Rhizome; the Sichuan Chinaberry Fruit is stir-fried Sichuan Chinaberry Fruit; the Frankincense and Myrrh are vinegar-processed Frankincense and Myrrh; the Atractylodes Macrocephalae is raw Atractylodes Macrocephalae; and the Licorice Root is prepared Licorice Root.

[0013] The traditional Chinese medicine composition of the present invention is prepared into a commonly used pharmaceutical formulation by adding pharmaceutically acceptable excipients or auxiliary ingredients, using the original medicinal material, water or organic solvent extract as the active ingredient.

[0014] The preparations mentioned therein are tablets, granules, pills, oral liquids, and capsules.

[0015] The present invention also provides a method for preparing the traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery, which includes the following steps:

[0016] a. Weigh the raw materials according to the specified weight ratio;

[0017] b. Grind into powder, or extract with water or organic solvents, and then add pharmaceutically acceptable excipients or auxiliary ingredients to prepare commonly used pharmaceutical formulations.

[0018] The present invention also provides the use of the aforementioned traditional Chinese medicine composition in the preparation of medicaments for treating and / or preventing intestinal adhesions after abdominal surgery.

[0019] The intestinal adhesions mentioned above are of the qi stagnation and blood stasis type.

[0020] The medication mentioned is for treating abdominal distension, abdominal pain, paroxysmal colic, shooting pain, stabbing pain, active bowel sounds, gurgling sounds, intestinal patterns, peristaltic waves, and constipation.

[0021] Postoperative intestinal adhesions can cause local microcirculatory disturbances, slow blood flow, leading to blood stasis, obstructed blood vessels, and pain. Essentially, it's a case of deficiency in the root and excess in the branch; the root deficiency is insufficient qi and blood, while the branch excess is qi stagnation and blood stasis. Therefore, qi stagnation and blood stasis are often the overall pathogenesis. Recurrent intestinal adhesions, coupled with emotional distress, liver qi stagnation, and qi imbalance, can further exacerbate the condition. Qi is the commander of blood; when qi flows smoothly, blood flows smoothly; when qi stagnates, blood stasis occurs, leading to pain. Therefore, while promoting blood circulation and removing blood stasis, supplementing with qi-regulating herbs can enhance the effects of promoting blood circulation and relieving pain. Simultaneously, liver qi stagnation and its rebellious nature can invade the spleen and stomach, causing dysfunction in their digestive functions. Therefore, supplementing with qi-tonifying, spleen-strengthening, stomach-harmonizing, and dampness-resolving herbs will yield better results.

[0022] The formula contains Sparganium rhizome and Curcuma rhizome, which break up blood stasis, promote qi circulation, eliminate stagnation, and relieve pain. Curcuma rhizome has a stronger qi-regulating effect, primarily targeting the qi level, and is good at breaking up blood stasis within the qi. Sparganium rhizome has a stronger blood-breaking effect, primarily targeting the blood level, and is good at breaking up qi stasis within the blood. The two work together synergistically, with a potent nature, effectively breaking up masses and accumulations, eliminating hardness, and relieving pain, specifically targeting stagnation in the intestines and stomach. Peach kernel is bitter and sweet in taste and neutral in nature, breaking up blood stasis and moistening dryness; Safflower is pungent and warm in nature, activating blood circulation and unblocking menstruation. The two work together synergistically, resolving stasis without being harsh, and moistening dryness without harming yin, together achieving the effects of activating blood circulation, removing blood stasis, unblocking meridians, and relieving pain. Combined with equal amounts of Sparganium rhizome and Curcuma rhizome, the four herbs work together to enhance the effects of activating blood circulation, removing blood stasis, promoting qi circulation, and relieving pain, and also provide moistening while breaking up stasis and nourishing while promoting circulation, making them the principal herbs.

[0023] Frankincense, being pungent and warm, is adept at promoting qi and blood circulation, relieving pain, and relaxing muscles; myrrh, being bitter and neutral, is adept at dispersing blood stasis, relieving pain, and promoting tissue regeneration. These two herbs work synergistically; after being processed with vinegar, they enter the liver meridian and blood level, further enhancing their blood-activating and pain-relieving effects. Vinegar-processed Corydalis activates blood, promotes qi circulation, and relieves pain; stir-fried Sichuan pepper soothes the liver, promotes qi circulation, and relieves pain. A large dose of white peony root (combined with licorice to form the White Peony and Licorice Decoction) is used to nourish yin with its sour and sweet properties, relieving spasms and pain, while simultaneously balancing the numerous pungent, warm, and drying herbs in the formula to prevent them from depleting blood and damaging yin. Used together with Corydalis and Sichuan pepper, it ensures that qi is promoted without depleting it, and blood stasis is resolved without harming the blood. The combined use of these five herbs enhances their effects of activating blood circulation, resolving blood stasis, soothing the liver, regulating qi, relieving spasms, and relieving pain, acting as assistant herbs.

[0024] The formula contains Poria cocos and Atractylodes macrocephala, which invigorate qi and strengthen the spleen, harmonize the stomach and resolve dampness, and alleviate the damage to the spleen and stomach caused by intestinal adhesions and various symptoms caused by spleen dysfunction. This allows the entire formula to both attack and tonify, remove blood stasis without harming the body's vital energy, and serve as adjuvant herbs.

[0025] Licorice harmonizes various pungent, warm, bitter, and cold herbs, serving as an adjuvant.

[0026] The beneficial effects of this invention are:

[0027] This invention relates to a traditional Chinese medicine composition for the treatment and / or prevention of intestinal adhesions after abdominal surgery, particularly for intestinal adhesions caused by qi stagnation and blood stasis, with significant efficacy. This invention also exhibits antagonistic effects against acute inflammation induced by xylene, with high and low dose groups showing superior efficacy compared to the medium dose group, and the high dose group demonstrating efficacy comparable to the chemical drug prednisolone acetate. Furthermore, it inhibits pain responses induced by acetic acid and hot plate therapy in mice, with the low dose showing significantly better effects than the medium and high dose groups. It promotes the propulsion of charcoal powder in the mouse intestine, with the high and low dose groups showing activity comparable to itopride, and significantly superior to the medium dose group. In rats with intestinal adhesion models, it reduces intestinal adhesions, alleviates intestinal tissue pathological damage to varying degrees, reduces fibrous tissue hyperplasia, and decreases the expression of TGF-β1 and TNF-α. Blood rheology and blood viscosity results show that this invention's traditional Chinese medicine composition has a certain effect in improving blood rheology and blood viscosity. In mice with intestinal adhesion models, it reduces intestinal adhesions, alleviates intestinal tissue pathological damage to varying degrees, and shows a trend towards reducing fibrous tissue hyperplasia. The results showed that the herbal composition of the present invention has anti-inflammatory, analgesic, intestinal motility-promoting, and anti-adhesion effects, the mechanism of which is partly related to the reduction of TGF-β1 and TNF-α expression. Furthermore, toxicological tests demonstrated that the herbal composition of the present invention is safer to use, providing a new medication option for clinical practice. Attached Figure Description

[0028] Figure 1 Histopathological images of rat intestinal tissue (HE staining);

[0029] Figure 2 Masson staining pathological images of rat intestinal tissue;

[0030] Figure 3 Image showing TGF-β1 expression;

[0031] Figure 4 Image showing TNF-α expression;

[0032] Figure 5 Pathological images of mouse intestinal tissue (HE staining);

[0033] Figure 6 This is a pathological image of mouse intestinal tissue stained with MASSON. Detailed Implementation

[0034] Example 1: Preparation of the traditional Chinese medicine composition of the present invention (hereinafter referred to as "Fukang Granules")

[0035] Prescription: Sparganium rhizome 10g, Curcuma rhizome 10g, Peach kernel 10g, Safflower 10g, White peony root 30g, Vinegar-processed Corydalis rhizome 10g, Fried Sichuan pepper 10g, Vinegar-processed frankincense 10g, Vinegar-processed myrrh 10g, Poria cocos 15g, Raw Atractylodes macrocephala 15g, Prepared licorice root 6g.

[0036] Indications: Promotes blood circulation, clears the meridians, regulates qi, and relieves pain. Used for abdominal and gynecological postoperative intestinal adhesions due to qi stagnation and blood stasis, characterized by paroxysmal abdominal pain, stabbing pain, colic, active bowel sounds, occasional visible intestinal loops, and difficulty in defecation. It can also be used for the prevention of abdominal and gynecological postoperative intestinal adhesions.

[0037] Dosage and administration: Decocted in water and taken orally, one dose for two days, three times a day.

[0038] Preparation process: The above twelve ingredients are decocted with water three times, each time for 1 hour. The decoctions are combined, filtered, and the filtrate is concentrated to an extract with a relative density of 1.15~1.30 (60~65℃). The extract is dried under reduced pressure, and an appropriate amount of dextrin and 4g of steviol glycosides are added. The extract is then pulverized, mixed, granulated, dried, and 1000g is obtained.

[0039] Example 2: Preparation of the traditional Chinese medicine composition of the present invention

[0040] Weigh the raw materials: 8g of Sparganium rhizome, 8g of Curcuma rhizome, 8g of Peach kernel, 8g of Safflower, 24g of White peony root, 8g of Corydalis rhizome, 8g of Sichuan pepper, 8g of Frankincense, 8g of Myrrh, 12g of Poria cocos, 12g of Atractylodes macrocephala, and 4.8g of Licorice root; decoct with water according to the method in Example 1, concentrate, and prepare an oral liquid.

[0041] Example 3: Preparation of the traditional Chinese medicine composition of the present invention

[0042] Weigh out the raw materials: 12g of Sparganium rhizome, 12g of Curcuma rhizome, 12g of Peach kernel, 12g of Safflower, 36g of White peony root, 12g of Corydalis rhizome, 12g of Sichuan pepper, 12g of Frankincense, 12g of Myrrh, 18g of Poria cocos, 18g of Atractylodes macrocephala, and 7.2g of Licorice root. Decoction with water according to the method in Example 1, concentrate, add starch to granulate, and prepare granules.

[0043] The beneficial effects of the present invention are demonstrated below through specific pharmacodynamic and toxicological tests.

[0044] Experimental Example 1: Pharmacodynamic Test of the Traditional Chinese Medicine Composition of the Present Invention

[0045] 1. Experimental Objective

[0046] Fukang granules have the effects of promoting blood circulation, clearing the meridians, regulating qi, and relieving pain. They are used for intestinal adhesions following abdominal and gynecological surgeries due to qi stagnation and blood stasis, characterized by paroxysmal abdominal pain, stabbing pain, colic, active bowel sounds, occasional visible intestinal loops, and difficulty defecating. They can also be used for the prevention of intestinal adhesions after abdominal and gynecological surgeries. This study evaluates the efficacy and some mechanisms of action of Fukang granules in terms of anti-inflammatory, analgesic, intestinal motility-promoting, blood-activating, and anti-adhesion effects, and explores its effective dosage and dose-effect relationship, providing a basis for clinical research.

[0047] 2. Experimental Materials

[0048] 2.1 Experimental Drugs

[0049] 2.1.1 Test Drugs

[0050] Fukang Granule dry extract powder: It is a brown-yellow powder, 1 g of the powder is equivalent to 4.83 g of crude drug respectively (batch number: 250301), stored in a -20°C refrigerator, and provided by Chengdu Baitai Pharmaceutical Co., Ltd. The clinically recommended daily dosage of Fukang Granule is 72.50 g of crude drug, calculated based on an adult body weight of 60 kg, which is 1.21 g of crude drug / kg / day. Prepare it into a corresponding concentration with pure water before use for standby.

[0051] The preparation method of Fukang Granule dry extract powder is as follows: Weigh 10 g of Trigonellae Rhizoma, 10 g of Curcumae Rhizoma, 10 g of Persicae Semen, 10 g of Carthami Flos, 30 g of Paeoniae Radix Alba, 10 g of Corydalis Rhizoma processed with vinegar, 10 g of Toosendan Fructus stir-fried, 10 g of Olibanum processed with vinegar, 10 g of Myrrha processed with vinegar, 15 g of Poria, 15 g of Atractylodis Macrocephalae Rhizoma crudum, 6 g of Glycyrrhizae Radix et Rhizoma praeparata cum melle, mix the above twelve medicinal materials, decoct with water three times, 1.0 hour for each time, combine the decoctions, filter, concentrate the filtrate to an extract with a relative density of 1.15~1.30 (60~65°C), dry under reduced pressure and pulverize to obtain the dry extract powder.

[0052] 2.1.2 Positive Drugs

[0053] Prednisone Acetate Tablets: Pujiang Xianju Pharmaceutical Co., Ltd., National Drug Approval Number: H31020593, batch number: LA24234. Specification: 5 mg / tablet.

[0054] Dexamethasone Acetate Tablets: Manufactured by Zhejiang Xianju Pharmaceutical Co., Ltd., National Drug Approval Number: H33020822, batch number: LB24193, specification: 0.75 mg / tablet.

[0055] Morphine Hydrochloride Sustained-release Tablets: Manufactured by Southwest Pharmaceutical Co., Ltd., batch number: 23030011, specification: 30 mg / tablet.

[0056] Aspirin Tablets: Manufactured by Shijiazhuang Pharmaceutical (CSPC) Group, batch number: 2862402094, specification: 100 mg / tablet.

[0057] Itopride Hydrochloride Capsules: Manufactured by Harbin Pharmaceutical Group General Pharmaceutical Factory, batch number: 240521, specification: 50 mg / capsule.

[0058] All the above drugs are prepared into corresponding concentrations with pure water immediately before use for standby.

[0059] 2.2 Experimental Animals and Experimental Environment

[0060] SPF-grade Kunming (KM) mice: Both male and female are used, with a body weight of 20~22 g, produced by Hunan Slack Jingda Experimental Animal Co., Ltd., production license number: SCXK (Xiang) 2021-0002.

[0061] SPF-grade Kunming (KM) mice: both male and female, weighted 14-20 g, produced by Miluo Branch of Hunan Slack Jingda Experimental Animal Co., Ltd., Production License No.: SCXK (Xiang) 2025-0004.

[0062] SPF-grade Kunming (KM) mice: all male, weighted 22-24 g, produced by Sichuan Weitong Lihua Experimental Animal Technology Co., Ltd., Production License No.: SCXK (Chuan) 2023-0040.

[0063] SPF-grade SD rats: all male, weighted 180-220 g, produced by Sichuan Weitong Lihua Experimental Animal Technology Co., Ltd., Production License No.: SCXK (Chuan) 2023-0040.

[0064] All animals were acclimatized for 3-5 days before the experiment. They were raised in the SPF barrier system of the Animal Experiment Center of Sichuan Academy of Chinese Medicine Sciences, the feeding conditions meet the relevant requirements of the barrier system, and the daily lighting time is 12 hours. Experimental Animal Use License No.: SYXK (Chuan) 2023-100.

[0065] Animal feed was provided by Beijing Huafukang Biotechnology Co., Ltd.

[0066] 2.3 Main Instruments

[0067] LD6100-1 electronic balance: Shenyang Longteng Electronics Co., Ltd., d=0.1 g;

[0068] MR203 electronic balance: Mettler-Toledo Instrument (Shanghai) Co., Ltd., d=1 mg;

[0069] BCE2241-1CCN electronic balance: Sartorius Scientific Instrument (Beijing) Co., Ltd., d=0.1 mg;

[0070] RB-201 intelligent hot plate instrument: Chengdu Taimeng Software Co., Ltd.;

[0071] 91404 (100 cm) straightedge: SATA, precision 1 mm;

[0072] Pipette, Eppendorf: 2-20 uL / 20-200 μL;

[0073] Assorted files: Jiangyin Shengda Diamond Products Factory, 3 mm×140 mm;

[0074] Absorbable surgical suture, sterile with curved needle (3 / 0, 90 cm), PGA polyglycolic acid absorbable material: Yuanlikang, Batch No.: 250910;

[0075] Leica 2016 rotary microtome: Leica, Germany;

[0076] JT-12S Automatic Dehydrator: Wuhan Junjie Electronics Co., Ltd.;

[0077] BMJ-A Tissue Embedding Machine: Changzhou Suburbs Zhongwei Electronic Instrument Factory;

[0078] PHY-Ⅲ Tissue Drying and Bleaching System: Changzhou Zhongwei Electronic Instruments Co., Ltd.;

[0079] SQS-600P Slide Scanning Imaging System: Shenzhen Shengqiang Technology Co., Ltd.

[0080] DHG-9148A Drying Oven: Shanghai Jinghong Experimental Equipment Co., Ltd.

[0081] A80400011 Restoration Instrument: Thermo Shandon Limited;

[0082] HY-5 Decolorizing Shaker: Shanghai Chengjie Instrument Equipment Co., Ltd.;

[0083] XK80-A Vortex Mixer: Jiangsu Xinkang Medical Equipment Co., Ltd.;

[0084] S1010E Handheld Centrifuge: SCLOGEX, USA.

[0085] 2.4 Reagents

[0086] Xylene: 500ml / bottle, Sichuan Kelun Pharmaceutical Co., Ltd., batch number: G220031804;

[0087] Glacial acetic acid: 500ml / bottle, Chengdu Changlian Chemical Reagent Co., Ltd., batch number: BCBH8982V;

[0088] Activated carbon (analytical grade, 200 mesh): 500g / bottle, Sinopharm Chemical Reagent Co., Ltd., batch number: 240210; Sodium carboxymethyl cellulose (CMC-Na) (chemically pure, viscosity 500-1000mPa・s): 500g / bottle, Shanghai Yuanye Biotechnology Co., Ltd., batch number: 240408;

[0089] Zoltil 50: Generic name: Telastatin-Zolazepam Injection, 50 mg / mL, Virbac, France, Batch No.: 9KM3A;

[0090] Zoltil 100: Generic name: Telastatin-Zolazepam Injection, 100 mg / mL, Virbac, France, Batch No.: 9KM3A;

[0091] 0.9% Sterile Sodium Chloride Injection (Physiological Saline for Injection): 100 mL / vial, Kelun Pharmaceutical (Sichuan Kelun Pharmaceutical Co., Ltd.), batch number: 25010806;

[0092] Sodium dihydrogen phosphate: 500g / bottle, Fuchen (Tianjin) Chemical Reagent Co., Ltd., batch number: 1010580101700;

[0093] Disodium hydrogen phosphate: 500g / bottle, Fuchen (Tianjin) Chemical Reagent Co., Ltd., batch number: 1010590101700; Formaldehyde (AR grade): 500ml / bottle, Sichuan Xilong Science Co., Ltd., batch number: 1340040101602.

[0094] Anhydrous ethanol (AR grade): 5L / barrel, Sinopharm Chemical Reagent Co., Ltd., batch number: 100092680;

[0095] Dewaxing agent / transparent agent: 2L / barrel, Wuxi Jiangyuan Industrial Technology and Trade Corporation, batch number: 240131;

[0096] Modified Masson's trichrome staining solution (potassium dichromate solution, Wergert's hematoxylin (solution b + solution c), Ponceau S, acid fuchsin solution, phosphomolybdic acid solution, aniline blue solution): 6*100ml / bottle, Wuhan Sewell Biotechnology Co., Ltd., batch number: G1006;

[0097] Hydrochloric acid (AR grade): 500ml / bottle, Chengdu Kelong Chemical Co., Ltd., batch number: 7647-01-0;

[0098] Neutral resin: 100g / bottle, Sinopharm Chemical Reagent Co., Ltd., batch number: 1004160;

[0099] Hematoxylin staining solution: 100g / bottle, Sigma Aldrich, batch number: H9627;

[0100] Eosin staining solution: 25g / bottle, Hefei Bomei Biotechnology Co., Ltd., batch number: YE2080; Dewaxing solution: Wuxi Jiangyuan Industrial Technology and Trade Co., Ltd., batch number: 240131;

[0101] Citrate buffer (dry powder): Servicebio, batch number: GA2307051;

[0102] PBS buffer (dry powder): Servicebio, batch number: G0002-2L;

[0103] Hydrogen peroxide: Xilong Scientific, batch number: SC2.0.0;

[0104] Bovine serum (BSA): Servicebio, batch number: GC305010;

[0105] DAB reagent kit: Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., batch number: ZL1-9018;

[0106] Hematoxylin staining solution: Beijing Bailingwei Technology Co., Ltd., batch number: LM10N13;

[0107] Hematoxylin differentiation solution: Servicebio, batch number: G1039;

[0108] Hematoxylin blue reversion solution: Servicebio, batch number: G1040;

[0109] Primary antibody (TGF-β1): Servicebio, batch number: GB15179;

[0110] Primary antibody (TNF-α): affinity, batch number: AF7014;

[0111] Secondary antibody (HRP-labeled goat anti-rabbit): Servicebio, batch number: GB22303;

[0112] Medical absorbent cotton: Sichuan Suining Kangda Sanitary Materials Co., Ltd., batch number: 20242140081.

[0113] 3. Experimental Methods and Results

[0114] 3.1 Anti-inflammatory effect

[0115] 3.1.1 Effect of xylene on ear swelling in mice

[0116] 3.1.1.1 Method

[0117] Fifty male Kunming mice were randomly divided into five groups of ten mice each, based on their body weight. These groups included a model control group, a positive control group (prednisolone acetate tablets, 10 mg / kg), and three dose groups of the test drug (high, medium, and low doses: 24.20, 12.10, and 6.05 g of raw drug / kg, equivalent to 20, 10, and 5 times the clinically recommended daily dose, respectively). The mice were administered the drug via gavage at equal volumes of different concentrations (the model control group received equal volumes of pure water). The gavage volume was 0.1 ml / 10 g B.W., once daily for five consecutive days. One hour after the last administration, 20 µl of xylene was evenly applied to both sides of the left ear of each mouse. Twenty minutes later, the mice were euthanized by cervical dislocation. Ear flaps from the same location on both ears were then punched using a 7 mm diameter punch and weighed on an electronic balance. The weight difference between the two ears, i.e., the degree of swelling, was calculated, and the swelling inhibition rate (%) for each group was calculated using the following formula. Experimental data were used... The results were expressed as t-tests or t' tests, with a significance level of α=0.05. Statistical analysis was performed using the PEMS 3.1 statistical software package (the same applies below).

[0118]

[0119] 3.1.1.2 Results

[0120] As shown in Table 1, compared with the model group, the high-dose Fukang granules group of mice showed a significant decrease in ear swelling. P <0.05), with an inhibition rate of 45.11%; the medium and low dose groups showed a decreasing trend in effect, with inhibition rates of 16.23% and 25.18%, respectively.

[0121] Table 1. Effect of Fukang Granules on Xylene-Induced Ear Swelling in Mice ( )

[0122]

[0123] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0124] 3.1.1.3 Conclusion

[0125] The results show that Fukang granules have an antagonistic effect on acute inflammation caused by xylene.

[0126] 3.2 Analgesic effect

[0127] 3.2.1 Effects on acetic acid-induced writhing response in mice

[0128] 3.2.1.1 Method

[0129] Fifty KM mice, half male and half female, were randomly divided into five groups (n=10 per group) according to weight and sex. These groups included a model control group, a positive control group (aspirin tablets, 100 mg / kg), and three dosage groups of the test drug (high, medium, and low-high doses: 24.20, 12.10, and 6.05 g of raw drug / kg, equivalent to 20, 10, and 5 times the clinically recommended daily dose, respectively). The drugs were administered via gavage at equal volumes of different concentrations (the model control group received equal volumes of pure water). The gavage volume was 0.1 ml / 10 g B.W., once daily for five consecutive days. One hour after the last administration, each group of animals was intraperitoneally injected with 0.1 ml / 10 g B.W. of 0.8% glacial acetic acid solution to induce pain. The number of writhing movements observed in the mice within 5–20 minutes after acetic acid injection was recorded, and the analgesia rate was calculated using the following formula. Experimental data were used as... The results were expressed as t-tests or t' tests, with a significance level of α=0.05.

[0130]

[0131] 3.2.1.2 Results

[0132] As shown in Table 2, compared with the model control group, the number of writhing movements in the low-dose Fukang granules group of mice was significantly reduced. P <0.01), with an analgesic rate of 23.95%.

[0133] Table 2. Effects of Fukang granules on acetic acid-induced writhing response in mice ( )

[0134]

[0135] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0136] 3.2.1.3 Conclusion

[0137] The results show that Fukang granules have an inhibitory effect on the writhing pain response induced by acetic acid in mice.

[0138] 3.3 Effects on charcoal propulsion in mice

[0139] 3.3.1 Method

[0140] Fifty KM mice, half male and half female, were randomly divided into five groups of ten mice each, based on body weight and sex. These groups included a model control group, a positive control group (itchopride hydrochloride capsules, 50 mg / kg), and three dosage groups (high, medium, and low doses of the test drug: 24.20, 12.10, and 6.05 g of raw drug / kg, equivalent to 20, 10, and 5 times the clinically recommended daily dose, respectively). The drugs were administered via gavage at equal volumes but different concentrations (the model control group received equal volumes of pure water). The gavage volume was 0.1 ml / 10 g B.W., once daily for three consecutive days. Animals were fasted for 16 hours prior to dissection, but water was permitted. One hour after the last administration, each group was gavage with 0.6 ml / mouse of a 10% activated charcoal suspension. Fifteen minutes later, the animals were euthanized by cervical dislocation. The small intestine was quickly removed, stretched into a straight line, and the total length of the small intestine from the pylorus to the ileocecal junction and the distance from the pylorus to the leading edge of the activated charcoal were measured. The charcoal propulsion rate (%) was calculated using the following formula. Experimental data used The results were expressed as t-tests or t' tests, with a significance level of α=0.05.

[0141]

[0142] Table 3. Effects of Fukang granules on the propulsion of charcoal powder in mice ( )

[0143]

[0144] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0145] 3.3.2 Results

[0146] As shown in Table 3, compared with the model control group, the dosage groups of Fukang granules significantly improved the charcoal powder propulsion rate, showing a statistically significant difference. P <0.05 or 0.01).

[0147] 3.3.3 Conclusion

[0148] The results show that Fukang granules have the effect of promoting intestinal peristalsis.

[0149] 3.4 Effects on the rat intestinal adhesion model

[0150] 3.4.1 Method

[0151] Sixty male SD rats were randomly divided into six groups of ten each, based on body weight. These groups included a sham-operated control group, a model control group, a positive control group (dexamethasone acetate tablets, 0.6 mg / kg), and three dosage groups of the test drug (19.36, 9.68, and 4.84 g of raw drug / kg, equivalent to 16, 8, and 4 times the clinically recommended daily dose, respectively). After acclimatization for five days, all rats were fasted for 12 hours, anesthetized with an intramuscular injection of 50 mg / kg of acetaminophen, and their abdominal hair was shaved. After disinfection with povidone-iodine, a 2-3 cm incision was made along the midline of the abdomen, from 2 cm below the sternum to 1 cm above the pubic symphysis, exposing the abdominal cavity. The skin and muscles were then incised sequentially. The cecum was gently removed, and the serosal layer was rubbed back and forth 10 times with a file to create a uniformly bleeding wound of 1.5cm × 1.5cm. The corresponding area on the opposite abdominal wall was rubbed in the same way to create a wound of the same area. The cecum was gently repositioned, and 0.5ml of sterile saline was dripped into the abdominal cavity per rat. The muscle layer and skin layer were sutured and disinfected separately. In the sham surgery group, the cecum was gently removed, and the corresponding abdominal wall was exposed without any manipulation. After exposure for 1 minute, the cecum was repositioned and sutured. The rats were fasted for 12 hours after surgery. After defecation, they were fed normally and housed separately for 5 days. Then, medication was started (the model control group and the sham surgery control group were given the same volume of pure water) once a day for 5 consecutive days. One hour after the last administration, the rats were anesthetized by intramuscular injection of Sutent 100 (50mg / kg). The abdomen was opened along the midline, and the formation of abdominal adhesions was observed and scored according to Table 4. The rats were then fixed in a lateral decubitus position on the operating table, and the intestinal segment was gently pulled out. The mesentery was laid flat on a transparent observation table and kept moist by dripping pre-warmed saline at 37°C. Transmitted light (10×-40× objective lens) was used to focus on mesenteric microvessels (arterioles, capillaries, venules) and measure vessel diameter (ingress and egress) and blood flow velocity. After observation, blood was collected from the abdominal aorta to measure blood rheology and viscosity. Corresponding intestinal and adhesion tissues were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE) to observe general lesions and score them according to Table 5. Masson staining was used to observe fibrosis and calculate the percentage of positive expression area. Immunohistochemistry was used to observe the expression of TNF-α and TGF-β1. Experimental data are expressed as scores and means. Results were analyzed using the rank-sum test with a significance level of α=0.05. Quantitative data were expressed as... The results were expressed as t-tests or t' tests, with a significance level of α=0.05.

[0152] Table 4 Scoring criteria for intestinal adhesion model (modified Phillips method)

[0153]

[0154] Table 5. Pathological Scoring Criteria for Intestinal Adhesions

[0155]

[0156] 3.4.2 Results

[0157] As shown in Table 6, all dosage groups of Fukang granules significantly reduced intestinal adhesion scores, with statistically significant differences. P <0.05 or 0.01).

[0158] Table 6. Effects of Fukang Granules on the Intestinal Tissue Adhesion Score in a Rat Intestinal Adhesion Model

[0159]

[0160] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0161] From Tables 7 to 10, Figure 1 It was found that in the sham-operated control group, the intestinal tissue mucosa, submucosa, muscularis propria, and serosa were relatively intact; the mucosal surface was covered with a single layer of columnar epithelium, with normal columnar epithelial cell morphology and no obvious degeneration, necrosis, or shedding; the colonic glands in the lamina propria were densely arranged, with a normal number of goblet cells, and a small number of scattered lymphocytes or macrophages were occasionally seen in the lamina propria; the thin layer of connective tissue in the submucosa contained abundant blood vessels; the muscularis propria was divided into inner circular and outer longitudinal muscles, with myenteric plexus visible between them. In the model control group, the intestine and adhesion tissue showed pathological changes such as myofiber degeneration and necrosis, inflammatory cell infiltration, fibrous tissue hyperplasia, neocapillary formation, and hemorrhage. The above-mentioned pathological damage in the intestinal tissue of each dose group of Fukang granules was reduced to varying degrees, showing a trend of reducing the pathological scores of inflammatory cell infiltration, fibrous tissue hyperplasia, and neocapillary formation, with the high-dose group showing the most significant reduction. P> 0.05).

[0162] Table 7. Effects of Fukang Granules on the Degree of Degeneration and Necrosis of Intestinal and Abdominal Wall Muscle Fibers in Rats

[0163]

[0164] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0165] Table 8. Effects of Fukang Granules on the Degree of Inflammatory Cell Infiltration in the Intestine and Abdominal Wall of Rats

[0166]

[0167] Note: Compared with the model control group (rank-sum test), *P<0.05, **P<0.01.

[0168] Table 9. Effects of Fukang Granules on the degree of proliferation of intestinal and abdominal wall muscle fiber tissue in rats.

[0169]

[0170] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0171] Table 10. Effects of Fukang Granules on the degree of neocapillary formation in the intestine and abdominal wall of rats.

[0172]

[0173] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0174] From Table 11, Figure 2 It was found that, compared with the model control group, the high-dose group of Fukang granules significantly reduced the percentage of positive expression area in fibrous tissue (P<0.05). The medium and low-dose groups showed a decreasing trend (P>0.05).

[0175] Table 11 Effects of Fukang Granules on the expression of fibrous tissue in the intestinal tissue of a rat intestinal adhesion model (Masson staining) )

[0176]

[0177] Note: Compared with the model control group: * P <0.05,** P <0.01.

[0178] From Table 12, Figure 3 and Figure 4 It was found that, compared with the model control group, the high and medium dose groups of Fukang granules could reduce the percentage of TGF-β1 expression area; the high dose group could reduce the percentage of TNF-α expression area, with significant differences (P<0.01). Figure 3 The arrows shown point to cells and regions where the target protein TGF-β1 is expressed.

[0179] Table 12 Effect of Fukang Granules on the Percentage of TGF-β1 and TNF-α Expression Area in Intestinal Tissue of Rat Intestinal Adhesion Model ( )

[0180]

[0181] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0182] As shown in Table 13, the different dosage groups of Fukang granules had no significant effect on mesenteric microcirculation.

[0183] Table 13 Effects of Fukang Granules on Mesenteric Microcirculation in a Rat Model of Intestinal Adhesion ( )

[0184]

[0185] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0186] As shown in Table 14, compared with the model control group, the high-dose group of Fukang granules increased the erythrocyte deformability index (TK); the high- and medium-dose groups decreased the erythrocyte aggregation index, with significant differences (P<0.05 or 0.01).

[0187] Table 14 Effects of Fukang Granules on Hemorheological Erythrocyte Index in Rat Intestinal Adhesion Model ( )

[0188]

[0189] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0190] As shown in Table 15, compared with the model control group, the medium-dose group of Fukang granules can reduce the low shear value of whole blood reduced viscosity, with a significant difference (P<0.05).

[0191] Table 15 Effects of Fukang Granules on Erythrocyte Sedimentation Rate and Blood Viscosity in a Rat Intestinal Adhesion Model ( )

[0192]

[0193] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0194] 3.4.3 Conclusion

[0195] The above results indicate that Fukang granules can alleviate intestinal adhesions in rats with intestinal adhesion model; reduce fibrous tissue hyperplasia; and alleviate intestinal pathological damage to varying degrees, showing a trend of reducing pathological scores for inflammatory cell infiltration, fibrous tissue hyperplasia, and neocapillary formation; and reducing the expression of TGF-β1 and TNF-α. These results suggest that Fukang granules have anti-inflammatory and anti-adhesion effects, partly related to the reduction of TGF-β1 and TNF-α expression. Blood rheology and blood viscosity results show that Fukang granules have a certain effect on improving blood rheology and blood viscosity.

[0196] 3.5 Effects on mouse intestinal adhesion model

[0197] 3.5.1 Method

[0198] Sixty male KM mice were randomly divided into six groups (n=10 per group) according to weight and sex: a sham-operated control group, a model control group, a positive control group (dexamethasone acetate tablets, 0.75 mg / kg), and three dose groups of the test drug (24.20, 12.10, and 6.05 g of raw drug / kg, equivalent to 20, 10, and 5 times the clinically recommended daily dose, respectively). After acclimatization for 4 days, all mice were fasted but allowed free access to water for 8 hours. After anesthesia with an intramuscular injection of 50 mg / kg of acetaminophen, the abdominal hair was shaved, and the area was disinfected with iodine. An incision of 0.8-1.2 cm was made along the midline of the abdomen from 0.5-1 cm below the sternum to 0.3-0.5 cm above the pubic symphysis, exposing the abdominal cavity. The cecum was gently removed, and the serosal layer was rubbed back and forth 10 times with a file to create a uniformly bleeding wound of 0.5cm × 0.5cm. The corresponding area on the opposite abdominal wall was rubbed in the same way to create a wound of the same area. The cecum was gently repositioned, and 0.1ml of sterile saline was dripped into the abdominal cavity per animal. The muscle layer and skin layer were sutured and disinfected separately. In the sham surgery group, the cecum was gently removed, and the corresponding abdominal wall was exposed without any manipulation. After exposure for 1 minute, it was repositioned and sutured. The animals were kept without food for 6 hours after surgery. After defecation, they were fed normally. They were then separated into standard cages and fed for 5 days before starting medication (the model control group and the sham surgery control group were given an equal volume of pure water) once a day for 5 consecutive days. One hour after the last administration, anesthesia was administered via intramuscular injection of 50 mg / kg of salbutamol. An laparotomy was performed along the midline of the abdomen to observe the formation of abdominal adhesions and score them according to Table 4. Then, corresponding intestinal and adhesion tissues were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE) to observe general lesions and score them according to Table 5. Masson staining was used to observe fibrosis and to measure and calculate the percentage of positive expression area. Experimental data are expressed as scores and means. Results were analyzed using the rank-sum test with a significance level of α=0.05. Quantitative data were expressed as follows: The results were expressed as t-tests or t' tests, with a significance level of α=0.05.

[0199] 3.5.2 Results

[0200] As shown in Table 16, all dosage groups of Fukang granules significantly reduced intestinal adhesion scores, with statistically significant differences. P <0.01).

[0201] Table 16 Effects of Fukang Granules on the Intestinal Tissue Adhesion Score in a Mouse Intestinal Adhesion Model

[0202]

[0203] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0204] From Tables 17 to 20, Figure 5 It was found that in the sham-operated control group, the intestinal tissue mucosa, submucosa, muscularis propria, and serosa were relatively intact; the mucosal surface was covered with a single layer of columnar epithelium, with normal columnar epithelial cell morphology and no obvious degeneration, necrosis, or shedding; the colonic glands in the lamina propria were densely arranged, with a normal number of goblet cells, and a small number of scattered lymphocytes or macrophages were occasionally seen in the lamina propria; the thin layer of connective tissue in the submucosa contained abundant blood vessels; the muscularis propria was divided into inner circular and outer longitudinal muscles, with myenteric plexus visible between them. In the model control group, pathological changes such as muscle fiber degeneration and necrosis, inflammatory cell infiltration, fibrous tissue hyperplasia, neocapillary formation, hemorrhage, and mineralization were observed. The severity of lesions was slightly reduced in the low-dose group of Fukang granules, somewhat reduced in the medium-dose group, and significantly reduced in the high-dose group and the positive control group. Among them, high and medium doses of Fukang granules can reduce the pathological score of myofibrosis degeneration and necrosis (P<0.05); low doses can reduce the pathological score of inflammatory cell infiltration (P<0.05); each dose group showed a trend of reducing the pathological score of fibrous tissue hyperplasia and new capillary formation (P>0.05).

[0205] Table 17 Effects of Fukang Granules on the Degree of Degeneration and Necrosis of Intestinal and Abdominal Wall Muscle Fibers in Mice

[0206]

[0207] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0208] Table 18 Effects of Fukang Granules on the Degree of Inflammatory Cell Infiltration in the Intestine and Abdominal Wall of Mice

[0209]

[0210] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0211] Table 19 Effects of Fukang Granules on the Degree of Intestinal and Abdominal Wall Muscle Fiber Proliferation in Mice

[0212]

[0213] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0214] Table 20 Effects of Fukang Granules on the Degree of Neocapillary Formation in the Intestine and Abdominal Wall of Mice

[0215]

[0216] Note: Compared with the model control group (rank-sum test), * P <0.05,** P <0.01.

[0217] From Table 21, Figure 6 It can be seen that the high and medium dose groups of Fukang granules showed a trend of reducing the percentage of positive expression area in fibrous tissue. P >0.05).

[0218] Table 21 Effects of Fukang Granules on the expression of fibrous tissue in the intestinal tissue of a mouse intestinal adhesion model (Masson staining) )

[0219]

[0220] Note: Compared with the model control group, * P <0.05,** P <0.01.

[0221] 3.5.3 Conclusion

[0222] The results above indicate that Fukang granules can alleviate intestinal adhesions in mice with an intestinal adhesion model; they also have varying degrees of mitigation effects on intestinal tissue pathological damage, reducing pathological scores for muscle fiber degeneration and necrosis and inflammatory cell infiltration, and showing a trend towards reducing pathological scores for fibrous tissue hyperplasia and neocapillary formation. These results suggest that Fukang granules possess anti-inflammatory and anti-adhesion effects.

[0223] 4. Experimental Conclusions

[0224] Fukang granules have an antagonistic effect on xylene-induced acute inflammation; they inhibit acetic acid-induced writhing pain in mice; they promote the propulsion of charcoal powder in the mouse intestine; and in rats with intestinal adhesion models, they can reduce intestinal adhesions, alleviate intestinal tissue pathological damage to varying degrees, reduce fibrous tissue hyperplasia, and decrease the expression of TGF-β1 and TNF-α. Blood rheology and blood viscosity results show that Fukang granules have a certain effect on improving blood rheology and blood viscosity. In mice with intestinal adhesion models, they can reduce intestinal adhesions, alleviate intestinal tissue pathological damage to varying degrees, and show a trend towards reducing fibrous tissue hyperplasia. These results indicate that Fukang granules have anti-inflammatory, analgesic, intestinal motility-promoting, and anti-adhesion effects, the mechanism of which is partly related to the reduction of TGF-β1 and TNF-α expression.

[0225] Experimental Example 2: Animal Toxicology Experiments of the Drug of the Present Invention

[0226] I. Acute toxicity test data and literature

[0227] 1. Experimental Objective

[0228] Based on the relevant provisions of the "Implementation Rules for the Filing Management of Traditional Chinese Medicine Preparations Prepared by Medical Institutions in Sichuan Province" and the "Technical Guidelines for Research on Preparations of Medical Institutions in Sichuan Province (Trial Version)," as well as the clinical application experience and indications of Fukang Granules, this experiment observed the toxic reactions and mortality of mice after a single oral administration of Fukang Granules within 14 days. The aim was to understand the acute toxicity of Fukang Granules, provide dosage references for pharmacodynamic studies, provide a reference basis for monitoring and treating clinical adverse reactions, and provide a basis for selecting dosages in long-term toxicity tests.

[0229] 2 Experimental Materials

[0230] 2.1 Experimental Drugs

[0231] Fukang Granules Dry Powder: Brownish-yellow powder, 1g of powder is equivalent to 4.83g of raw herb, supplied by Chengdu Byte Pharmaceutical Co., Ltd. Batch No.: 250301, store at -20℃.

[0232] Preparation method: Accurately weigh 67.0g of the test drug, add an appropriate amount of pure water and grind evenly, and finally adjust the volume to 100ml to obtain the maximum concentration 67% suspension (100ml of suspension contains 67g of drug powder, and its viscosity is limited to passing through a No. 16 g gavage needle) for test use.

[0233] 2.2 Laboratory animals and experimental environment

[0234] Kunming mice (KM), weighing 16-18g, 40 mice, half male and half female, SPF grade, provided by Miluo Branch of Hunan Slake Jingda Experimental Animal Co., Ltd., Production License No.: SCXK (Xiang) 2025-0004.

[0235] All animals were acclimatized for 4 days before the animal experiment. They were raised in the SPF barrier system of the Animal Experimental Center of Sichuan Academy of Chinese Medicine Sciences, and the feeding conditions meet the relevant requirements of the barrier system, with 12 hours of light per day. License number for the use of experimental animals: SYXK (Chuan) 2023-100.

[0236] Animal feed was provided by Beijing Huafukang Biotechnology Co., Ltd.

[0237] 2.3 Main Instruments

[0238] LD6100-1 electronic analytical balance: d=0.1 g, manufactured by Shenyang Longteng Electronics Co., Ltd.

[0239] 3 Experimental Methods and Results

[0240] 3.1 Experimental Methods

[0241] It can be known from the pre-test that no animal death was observed when mice were given the test drug by single intragastric administration at the maximum administration dosage, so LD 50 and the maximum tolerated dose cannot be determined, therefore, the maximum administration dosage was determined.

[0242] The experimental design is shown in Table 22. Forty KM mice were acclimatized for 4 days and randomly divided into the Fukang Granule group and the blank control group according to body weight, with 20 mice in each group and half male and half female. After 16 hours of fasting with free access to water, animals in each group were given single intragastric administration at the maximum concentration of Fukang Granule dry extract powder (67% suspension, 100 ml of the suspension contains 67 g of medicinal powder, and its viscosity is limited by being able to pass through No. 16 intragastric needle) and the maximum volume (0.4 ml / 10 g B.W.). The control group was given an equal volume of pure water. The day of intragastric administration was recorded as Day 1. Observations were performed immediately after administration, and then continued for 14 consecutive days. The general condition and death of mice were observed every day. On Day 14, animals were euthanized, systematic autopsy was performed, and the size, shape, color and texture of each main organ were observed. When macroscopic lesions were visible, corresponding histopathological examination was performed. Body weights were weighed on Day 1 (before administration), Day 3, Day 7 and Day 14. The t-test or t'-test was used to compare the significance of differences between the two groups, with the test level α=0.05. Statistical analysis was performed using the PEMS 3.1 statistical software package.

[0243] Table 22 Design of acute toxicity test of Fukang Granule in mice

[0244]

[0245] 3.2 Experimental Results

[0246] 3.2.1 Observation of General Conditions

[0247] No animal death occurred during the test.

[0248] In the Fukang granule group, animals showed reduced movement 30 minutes after administration, which gradually recovered 2 hours later. No other abnormalities were observed in reflexes, eyelid signs, cardiovascular signs, salivation, pain sensation, muscle tone, gastrointestinal signs, urinary system, or skin.

[0249] Observations from Day 2 to Day 14 showed no abnormalities in the animal's appearance, behavior, excretion, secretion, or response to stimuli.

[0250] 3.2.2 Weight Changes

[0251] The results are shown in Tables 23-25.

[0252] Table 23. Weight Changes in Male Animals ( )

[0253]

[0254] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0255] Table 24 Body Weight Changes in Female Animals ( )

[0256]

[0257] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0258] Table 25. Weight Changes in Male and Female Animals ( )

[0259]

[0260] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0261] 3.2.3 Gross Anatomy

[0262] After gross dissection, no abnormal changes were observed in the size, shape, color, or texture of the major organs and tissues upon visual inspection.

[0263] 4. Experimental Conclusions

[0264] The recommended daily dose of Fukang granules for adults is 72.50g of raw herb. For an adult weighing 60kg, the daily oral dose of Fukang granules is 1.21g of raw herb / kg. Under the conditions of this experiment, the maximum dose of Fukang granules administered to KM mice via a single gavage was 26.80g / kg, or 129.44g of raw herb / kg, equivalent to 107 times the recommended daily dose.

[0265] II. Long-term toxicity test data

[0266] 1. Experimental Objective

[0267] Based on the relevant provisions of the "Implementation Rules for the Filing Management of Traditional Chinese Medicine Preparations Prepared by Medical Institutions in Sichuan Province" and the "Technical Guidelines for Research on Preparations of Medical Institutions in Sichuan Province (Trial Version)," as well as the clinical application experience and indications of Fukang Granules, this experiment plans to set up high-, medium-, and low-dose groups of Fukang Granules and a blank control group. The toxic reactions of animals after 91 days of continuous administration and the recovery of animals after 28 days of drug withdrawal will be observed, and various observation indicators will be detected to understand the toxic reactions of this preparation to animals and provide a reference for safe clinical drug use.

[0268] 2. Experimental Materials

[0269] 2.1 Experimental Drugs

[0270] Fukang Granules (dry powder): A brownish-yellow powder. 1 gram of powder is equivalent to 4.83 grams (batch number: 250301) and 4.63 grams (batch number: 250401) of the raw medicinal material, respectively. Store at -20℃. Provided by Chengdu Baiter Pharmaceutical Co., Ltd. The clinically recommended daily dosage of Fukang Granules is 72.50g of raw medicinal material / day. For an adult weighing 60kg, this equates to 1.21g of raw medicinal material / kg / day.

[0271] Preparation of test drug: Weigh an appropriate amount of Fukang granules dry powder daily, add an appropriate amount of pure water and grind, and finally dilute with pure water to a final concentration of 75.05% (w / v), 37.53% (w / v), 18.76% (w / v) (batch number: 250301) or 78.29% (w / v), 39.15% (w / v), 19.57% (w / v) (batch number: 250401) suspension for use in the high, medium and low dose groups of the long-term toxicity test of Fukang granules.

[0272] 2.2 Laboratory animals and experimental environment

[0273] Laboratory animals: Sprague Dawley (SD) rats, weighing 100-120g, 120 rats, half male and half female, SPF grade, provided by Hunan Slack Jingda Laboratory Animal Co., Ltd., production license number: SCXK (Xiang) 2021-0002.

[0274] All animals were acclimatized for one week before all animal experiments. They were raised in the SPF barrier system of the Animal Experimental Center of Sichuan Academy of Chinese Medicine Sciences, and the feeding conditions met the relevant requirements of the barrier system, with a daily light duration of 12 hours. The license number for the use of experimental animals: SYXK (Chuan) 2023-100.

[0275] Animal feed was provided by Beijing Huafukang Biotechnology Co., Ltd.

[0276] 2.3 Main Instruments

[0277] Model 7180 automatic biochemical analyzer (manufactured by Hitachi High-Technologies Corporation, Japan);

[0278] BC-6900 automatic blood cell analyzer (Mindray Biomedical Electronics Co., Ltd., Shenzhen);

[0279] ExC810 automatic coagulation analyzer (Mindray Biomedical Electronics Co., Ltd., Shenzhen);

[0280] HM340E rotary microtome (Epredia Scientific Equipment Manufacturing (Shanghai) Co., Ltd.);

[0281] PQFA automatic tissue dehydrator (Presing (Changzhou) Medical Device Co., Ltd.);

[0282] YB-7LF tissue embedding machine (Xiaogan Yaguang Medical Electronic Technology Co., Ltd.);

[0283] YT-7FB pathological tissue floating and baking instrument (Xiaogan Yaguang Medical Electronic Technology Co., Ltd.);

[0284] OLYMPUS CX43 biological microscope (Evident Optical Technology (Guangzhou) Co., Ltd.);

[0285] LD6100-1 electronic balance (d=0.1 g, Shenyang Longteng Electronics Co., Ltd.);

[0286] MR203 electronic balance (d=1 mg, Mettler-Toledo Instruments (Shanghai) Co., Ltd.).

[0287] 2.4 Main Reagents

[0288] The names, manufacturers and batch numbers of the reagents involved in this test are shown in Table 26.

[0289] Table 26 Table of Main Reagents

[0290]

[0291] 3 Experimental Methods and Results

[0292] 3.1 Experimental Methods

[0293] 3.1.1 Dosage and Grouping

[0294] As shown in Table 27, there were three treatment groups: a high-dose group of Fukang granules (72.50 g raw drug / kg), a medium-dose group (36.25 g raw drug / kg), a low-dose group (18.13 g raw drug / kg), and a blank control group.

[0295] Table 27 Dosage and Grouping Table

[0296]

[0297] 3.1.2 Administration method

[0298] The test drug solutions of different concentrations but the same volume were administered by gavage, while the blank control group was administered the same volume of pure water. The administration volume was 20 ml / kg B.W. / time, administered once daily between 8-11 am, 6 days a week for 91 consecutive days, followed by a 28-day observation period without administration of the test drug or pure water (control group). Body weight and feed consumption were measured weekly.

[0299] 3.1.3 Testing methods and testing time for each testing indicator

[0300] 3.1.3.1 Detection and observation methods for each indicator

[0301] (1) General observation: Before and after daily administration, observe the appearance, behavior, shape and color of urine and feces, and any abnormal secretions from each natural orifice of each animal in each dosage group. Weigh the animal once before administration and weigh the animal and measure its daily feed consumption once a week during administration.

[0302] (2) Peripheral blood count: Blood was collected from the abdominal aorta of the animal after anesthesia, and anticoagulated with EDTA. The following values ​​were measured: WBC (white blood cells), NEU# (neutrophil count), NEU% (neutrophil percentage), LYMPH# (lymphocyte count), LYMPH% (lymphocyte percentage), RBC (red blood cells), HGB (hemoglobin), HCT (hematocrit), MCV (mean corpuscular volume), MCH (mean corpuscular hemoglobin), MCHC (mean corpuscular hemoglobin concentration), RET# (reticulocyte count), RET% (reticulocyte percentage), and PLT (platelets).

[0303] (3) Blood biochemistry tests: Blood was drawn from the abdominal aorta as described above, serum was separated, and the following parameters were tested: ALT (alanine aminotransferase), AST (aspartate aminotransferase), TP (total serum protein), ALB (serum albumin), TBIL (total bilirubin), ALP (alkaline phosphatase), GLU (blood glucose), Urea (urea), Creatinine (creatinine), TC (total cholesterol), TG (triglycerides), CK (creatine kinase), and K. + (serum potassium), Na + (serum sodium), Cl - (Serium chloride).

[0304] (4) Coagulation function test: Blood was drawn from the abdominal aorta as described above, anticoagulated with sodium citrate, and prothrombin time (PT) and activated partial thromboplastin time (APTT) were measured.

[0305] (5) Systematic autopsy: After the animal is killed, a comprehensive and systematic autopsy is performed in accordance with the "Operating Procedure for Autopsy of Experimental Animals". The appearance and texture of the major organs are observed. The organs such as brain, heart, liver, spleen, lungs, kidneys, adrenal glands, thymus, testes, epididymis, uterus, and ovaries are weighed and the organ coefficients are calculated.

[0306] (6) Histopathological examination: Brain (cerebrum, cerebellum, brainstem), spinal cord (cervical, thoracic, lumbar segments), pituitary gland, thymus, thyroid gland, parathyroid gland, esophagus, salivary glands, stomach, small intestine, large intestine, liver, kidneys, adrenal glands, spleen, pancreas, trachea, lungs, aorta, heart, epididymis, testes, ovaries, uterus, prostate, breast, sciatic nerve, bladder, bone marrow, lymph nodes, etc. are taken, fully fixed in 10% formalin, routinely embedded in paraffin, sectioned, stained with HE, observed under a light microscope and photographed.

[0307] 3.1.3.2 Detection Time

[0308] As shown in Table 28.

[0309] Table 28 Testing Items and Testing Schedule

[0310]

[0311] Note: The interval between weighing body weight and feed consumption is approximately 7 days. Please refer to the relevant tables below for specific times.

[0312] 3.1.4 Statistical Methods

[0313] Experimental data used The results were expressed as t-tests or t' tests, with a significance level of α=0.05. Statistical analysis was performed using the PEMS 3.1 statistical software package.

[0314] 3.2 Experimental Results

[0315] 3.2.1 General Observation

[0316] Before and for 91 days after drug administration, the animals in each group showed no abnormalities in appearance, behavior, coat luster, and were visually normal in urine and feces. Their respiratory, nervous, and digestive systems were also normal. There were no abnormal secretions from the mouth, eyes, nose, ears, genitals, or other natural orifices. After 28 days of observation following drug withdrawal, no abnormalities were observed in either the drug-treated or control groups.

[0317] Feed consumption: As animals age and gain weight, feed consumption increases, while the daily feed consumption per unit body weight decreases. Compared with the blank control group, there was no significant difference in feed consumption at each time point among the different dosage groups of Fukang granules. P >0.05). See Table 29 for details.

[0318] Body weight: Compared with the blank control group, the female rats in the medium-dose group of Fukang granules had a higher body weight on day 21, and the difference was statistically significant. P <0.05%, and there were no statistically significant differences between the other dosage groups and the blank control group at other time points. P >0.05); Compared with the blank control group, the body weight of male rats in the high- and medium-dose groups of Fukang granules decreased on days 14 and 28, and the differences were statistically significant. P <0.05%, and there were no statistically significant differences between the other dosage groups and the blank control group at other time points. P >0.05); There was no statistically significant difference in body weight (female + male) of rats in each of the Fukang granule treatment groups compared with the blank control group. P >0.05). See Tables 30-32 for details.

[0319] Table 29 Daily feed consumption of rats in each group (g / 100g BW)

[0320]

[0321] Note: n=30 before and during drug administration, n=10 during the drug withdrawal period. Compared with the blank control group, * P <0.05,** P <0.01.

[0322] Table 30 Weight changes (g) after 91 days of drug administration and 28 days after drug withdrawal (♀) )

[0323]

[0324] Note: Before and during drug administration, n=15 animals; during the withdrawal observation period, n=5 animals (of which, 10 animals were sacrificed for observation). Compared with the blank control group, * P <0.05,**P <0.01.

[0325] Table 31 Weight Changes (g) after 91 Days of Drug Administration and 28 Days After Drug Discontinuation (♂) )

[0326]

[0327] Note: Before and during drug administration, n=15 animals; during the withdrawal observation period, n=5 animals (of which, 10 animals were sacrificed for observation). Compared with the blank control group, * P <0.05,** P <0.01.

[0328] Table 32 Weight changes (g) after 91 days of drug administration and 28 days of drug withdrawal (♂+♀) )

[0329]

[0330] Note: Before and during drug administration, n=30; during the withdrawal observation period, n=10 (20 were euthanized for observation, 10 each of sexes). Compared with the blank control group, * P <0.05,** P <0.01.

[0331] 3.2.2 Peripheral blood count and coagulation function tests

[0332] After 91 days of administration, compared with the blank control group, the MCV, MCH, RET#, and RET% of the high, medium, and low dose groups of Fukang granules were all increased. P <0.01), RBC decreased ( P (<0.05 or 0.01), the difference is statistically significant. See Table 33 for details.

[0333] After 28 days of observation following drug withdrawal, there were no statistically significant differences in peripheral blood counts and coagulation function among the various dosage groups of Fukang granules compared to the blank control group. P >0.05). See Table 34 for details.

[0334] Table 33 Peripheral blood count and coagulation function test results after 91 days of drug administration (♂+♀) )

[0335]

[0336] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0337] Table 34 Peripheral blood count and coagulation function test results 28 days after drug withdrawal (♂+♀) )

[0338]

[0339] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0340] 3.2.3 Blood biochemistry tests

[0341] After 91 days of administration, compared with the blank control group, the high-dose group of Fukang granules showed a decrease in TG ( P <0.01, the medium-dose group showed a decrease in GLU, CK, and Cl ( P <0.01 or P <0.05, low-dose group showed decreased GLU, Crea, and Cl ( P <0.01 or P The difference was <0.05, which was statistically significant. See Table 35 for details.

[0342] After 28 days of observation following drug withdrawal, compared with the blank control group, the high-dose Fukang granules group showed a decrease in TBIL and Crea. P <0.05%, Na and Cl levels increased in the medium-dose group ( P <0.05), Crea decreased ( P <0.01); TBIL decreased in the low-dose group ( P The difference was <0.05, which was statistically significant. See Table 36 for details.

[0343] Table 35. Blood Biochemistry Results After 91 Days of Drug Administration (♂+♀) )

[0344]

[0345] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0346] Table 36 Blood Biochemistry Test Results 28 Days After Medication Discontinuation (♂+♀) )

[0347]

[0348] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0349] 3.2.4 Systemic autopsy and organ coefficients

[0350] 3.2.4.1 Anatomical observation

[0351] After 91 days of administration, no abnormalities were observed in the arrangement, color, or texture of the animal's organs and tissues.

[0352] After 28 days of observation following drug withdrawal, no abnormalities were observed in the arrangement, color, or texture of the animal's organs and tissues.

[0353] 3.2.4.2 Organ Coefficient

[0354] After 91 days of administration, the epididymal coefficient in the low-dose group of male rats decreased, and the difference was statistically significant compared with the blank control group. P <0.05); The kidney coefficient of female rats in the low-dose group was increased, and the difference was statistically significant compared with the blank control group. P <0.05). See Tables 37 and 38 for details.

[0355] After 28 days of observation following drug withdrawal, there were no statistically significant differences in the organ coefficients of rats in each of the Fukang granule treatment groups compared with the blank control group. P >0.05). See Tables 39 and 40 for details.

[0356] Table 37 Organ coefficients after 91 days of drug administration (organ g / 100g body weight) (♂) )

[0357]

[0358] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0359] Table 38 Organ coefficients after 91 days of drug administration (organ g / 100g body weight) (♀) )

[0360]

[0361] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0362] Table 39 Organ coefficient table 28 days after drug withdrawal (organ g / 100g body weight) (♂) )

[0363]

[0364] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0365] Table 40 Organ coefficient table 28 days after drug withdrawal (organ g / 100g body weight) (♀) )

[0366]

[0367] Note: Compared with the blank control group, * P <0.05,** P <0.01.

[0368] 3.2.4.5 Histopathological examination

[0369] No significant histopathological changes caused by the test drug were observed in organs including the brain (cerebrum, cerebellum, brainstem), spinal cord (cervical, thoracic, and lumbar segments), pituitary gland, thymus, thyroid gland, parathyroid gland, esophagus, salivary glands, stomach, small intestine, large intestine, liver, kidneys, adrenal glands, spleen, pancreas, trachea, lungs, aorta, heart, epididymis, testes, ovaries, uterus, prostate, breast, sciatic nerve, bladder, bone marrow, and lymph nodes after 91 days of drug administration and 28 days of observation following drug withdrawal.

[0370] 3.3 Summary

[0371] 3.3.1 General Observation

[0372] Before and for 91 days after drug administration, the animals in each group showed no abnormalities in appearance, behavior, coat luster, and were visually normal in urine and feces. Their respiratory, nervous, and digestive systems were also normal. There were no abnormal secretions from the mouth, eyes, nose, ears, genitals, or other natural orifices. After 28 days of observation following drug withdrawal, no abnormalities were observed in either the drug-treated or control groups.

[0373] 3.3.2 Feed consumption and body weight

[0374] Feed consumption: As animals age and gain weight, feed consumption increases, while the daily feed consumption per unit body weight decreases. Compared with the blank control group, there was no significant difference in feed consumption among the different dosage groups of Fukang granules during the administration period and the observation period after drug withdrawal. P >0.05).

[0375] Body weight: The body weight of male rats in the high- and medium-dose groups decreased on days 14 and 28. P <0.05%, but there was no obvious dose-response relationship, and the reduction was not significant, so it was considered to have no significant biological significance. The female rats in the medium-dose group showed an increase in body weight on day 21 ( P <0.05%, indicating no significant dose-response relationship and a small increase, therefore considered to have no significant biological significance. There was no significant difference in overall weight between male and female animals during the drug administration period and the observation period after drug withdrawal. P >0.05). After 28 days of observation following drug withdrawal, there was no significant difference in weight gain among the groups.

[0376] 3.3.3 Peripheral blood count and coagulation function tests

[0377] After 91 days of administration, the levels of MCV, MCH, RET#, and RET% increased in the high, medium, and low dose groups of Fukang granules. P <0.01), RBC decreased ( P <0.05 or P <0.01). The RBC reduction rates were 6.69%, 7.18%, and 5.72%, respectively; the MCV increase rates were 5.07%, 4.95%, and 4.87%, respectively; and the MCH increase rates were 3.85%, 4.63%, and 4.14%, respectively. The changes in these indicators were all small, and there was no dose-response relationship. All changes recovered after drug discontinuation. Combined with normal bone marrow pathological examination, this indicates that the above changes had no significant biological significance. The RET# increase rates were 29.22%, 27.27%, and 48.05%, respectively; and the RET% increase rates were 38.50%, 3.36%, and 58.82%, respectively. The changes were all large, but there was no dose-response relationship. The increase in RET# and RET% may be related to the feedback effect of RBC reduction leading to increased bone marrow hematopoietic function, which is a physiological feedback regulation of the body. Therefore, it is considered that no biologically significant changes in peripheral blood counts were observed due to the drug.

[0378] 3.3.4 Blood biochemistry tests

[0379] After 91 days of administration, the TG level in the high-dose group of Fukang granules decreased ( P <0.01, the medium-dose group showed a decrease in GLU, CK, and Cl ( P <0.01 or P <0.05, low-dose group showed decreased GLU, Crea, and Cl ( P <0.01 or P <0.05%. After 28 days of observation following drug withdrawal, TBIL and Crea levels decreased in the high-dose group of Fukang granules (<0.05). P <0.05%, Na and Cl levels increased in the medium-dose group ( P <0.05), Crea decreased ( P <0.01); TBIL decreased in the low-dose group ( P <0.05). The changes in the above indicators were small, with no obvious dose-response relationship, and all returned to normal after drug withdrawal. Decreases in CK, TG, Crea, Urea, and TBIL generally have no significant toxicological significance. The reduction rates of GLU in the medium-dose and low-dose groups were 17.42% and 15.42%, respectively, with small decreases and no obvious dose-response relationship; therefore, the changes had no significant biological significance. The changes in Cl and Na were extremely small, with no obvious dose-response relationship; therefore, they were considered to have no significant biological significance. Therefore, it was concluded that no drug-induced changes in blood biochemistry were biologically significant.

[0380] 3.3.5 Systemic autopsy and organ coefficients

[0381] After 91 days of drug administration and 28 days of drug withdrawal, autopsies were performed on the organs and tissues of the animals in both the drug administration group and the control group, and no abnormal lesions were observed to the naked eye.

[0382] After 91 days of administration, the epididymal coefficient in the low-dose group of male rats decreased ( P <0.05%, the kidney coefficient was increased in the low-dose group of female rats ( P <0.05), indicating no significant dose-response relationship; all results returned to normal after drug discontinuation. Combined with the absence of significant histopathological changes in the epididymis and kidneys in the high-dose group, it was considered to have no significant toxicological significance. Therefore, it was concluded that no biologically significant changes in organ coefficients caused by the drug were observed.

[0383] 3.3.6 Histopathological examination

[0384] No obvious organ tissue pathological changes caused by the drug were observed.

[0385] 4. Experimental Conclusions

[0386] Under the conditions of this experiment, Fukang granules were administered orally at doses of 72.50, 36.25, and 18.13 g of raw drug / kg (equivalent to 60, 30, and 15 times the clinically recommended daily dose, respectively) for 91 days, followed by a 28-day observation period after drug withdrawal. No clinically significant biological changes were observed in the animals' general condition, feed consumption, body weight, peripheral blood count, blood biochemistry, major organ coefficients, or histopathology.

Claims

1. A traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: 8-12 parts of Sparganium rhizome, 8-12 parts of Curcuma rhizome, 8-12 parts of Peach kernel, 8-12 parts of Safflower, 24-36 parts of White peony root, 8-12 parts of Corydalis rhizome, 8-12 parts of Sichuan Chinaberry, 8-12 parts of Frankincense, 8-12 parts of Myrrh, 12-18 parts of Poria cocos, 12-18 parts of Atractylodes macrocephala, and 4.8-7.2 parts of Licorice root.

2. The traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery according to claim 1, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: 10 parts of Sparganium rhizome, 10 parts of Curcuma rhizome, 10 parts of Peach kernel, 10 parts of Carthamus tinctorius, 30 parts of Paeonia lactiflora, 10 parts of Corydalis yanhusuo, 10 parts of Melia toosendan, 10 parts of Boswellia carterii, 10 parts of Commiphora myrrha, 15 parts of Poria cocos, 15 parts of Atractylodes macrocephala, and 6 parts of Glycyrrhiza uralensis.

3. The traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery according to claim 1 or 2, characterized in that: The Corydalis Rhizome mentioned is vinegar-processed Corydalis Rhizome; the Sichuan Chinaberry Fruit mentioned is stir-fried Sichuan Chinaberry Fruit; the Frankincense and Myrrh mentioned are vinegar-processed Frankincense and Myrrh; the Atractylodes Macrocephalae mentioned is raw Atractylodes Macrocephalae; and the Licorice Root mentioned is prepared Licorice Root.

4. The traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery according to claim 1 or 2, characterized in that: It is a pharmaceutically commonly used preparation made from the original medicinal material, water or organic solvent extract of the active ingredient, and with the addition of pharmaceutically acceptable excipients or auxiliary ingredients.

5. The traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery according to claim 4, characterized in that: The preparations mentioned are tablets, granules, pills, oral liquids, and capsules.

6. A method for preparing the traditional Chinese medicine composition for treating and / or preventing intestinal adhesions after abdominal surgery as described in any one of claims 1-5, characterized in that: It includes the following steps: a. Weigh the raw materials according to the specified weight ratio; b. Grind into powder, or extract with water or organic solvents, and then add pharmaceutically acceptable excipients or auxiliary ingredients to prepare commonly used pharmaceutical formulations.

7. Use of the traditional Chinese medicine composition according to any one of claims 1-5 in the preparation of a medicament for treating and / or preventing intestinal adhesions after abdominal surgery.

8. The use according to claim 7, characterized in that: The intestinal adhesions mentioned are of the qi stagnation and blood stasis type.

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