Traditional Chinese medicine composition for treating portal vein and hepatic sinus vascular diseases and application of traditional Chinese medicine composition

Abstract

The invention relates to a traditional Chinese medicine composition for treating portal vein and hepatic sinus vascular diseases. The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 13-17 parts of astragalus membranaceus, 13-17 parts of angelica sinensis and 4-8 parts of leech. The invention also provides application of the traditional Chinese medicine composition in preparation of medicines for treating portal vein and hepatic sinus vascular diseases. According to the invention, an SD (Sprague Dawley) rat is fed with selenium-rich diet, meanwhile, a model is made according to a chemotherapy regimen of FOLFOX intravenously injected into the tail of the rat every week, the model is made continuously for 8 weeks, a PSVD animal model is copied, the modified Chinese angelica blood replenishing decoction is fed for intervention while the model is made, and simvastatin is taken as a positive control. Results show that the modified Chinese angelica blood replenishing decoction can significantly reduce the portal vein pressure of rats and improve related expressions of liver microangiopathy. The liver index is obviously reduced, and the spleen index is improved. Liver tissue pathological reticular staining and Japanese stellate red staining show that both the liver tissue and the Japanese stellate red staining are relieved.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically, to a traditional Chinese medicine composition for treating porto-sinusoidal vascular disorder and its application. Background Art

[0002] Porto-sinusoidal vascular disorder (PSVD) is a newly proposed disease name in recent years and is an important supplement to idiopathic non-cirrhotic portal hypertension (INCPH). INCPH is a pre-sinusoidal PH disease in the liver and has become increasingly common clinically in recent years. Its significant feature is that the clinical symptoms of PH (such as splenomegaly, esophageal and gastric fundus varices, etc.) are significant but there is no liver cirrhosis. Traditional Chinese medicine usually conducts syndrome differentiation and treatment based on the clinical manifestations and symptoms of patients, and may classify it into the categories of "hypochondriac pain", "accumulation", "abdominal distension", etc. Traditional Chinese medicine believes that the occurrence of this disease may be related to factors such as liver qi stagnation, qi stagnation and blood stasis, and damp-heat accumulation.

[0003] Chinese patent document CN105617255A discloses a traditional Chinese medicine for treating liver cirrhosis of portal vein type with liver depression and spleen deficiency. Its special feature is that the components include: 15 grams each of angelica and turtle shell, 9 grams each of bupleurum, white peony root, cyperus rotundus, curcuma aromatica, atractylodes macrocephala, peach kernel, safflower, and chicken gizzard membrane, and 12 grams of codonopsis pilosula. It solves the problem of the high cost of using western medicine to control liver cirrhosis and is very suitable for patients with liver cirrhosis of portal vein type with liver depression and spleen deficiency.

[0004] Chinese patent document CN106474302A discloses a traditional Chinese medicine for treating liver cirrhosis of portal vein type with blood stasis stagnation. Its special feature is that the components include: 9 grams each of peach kernel, red peony root, safflower, and fructus aurantii, 12 grams each of angelica and cyperus rotundus, 18 grams of turtle shell, and 30 grams of salvia miltiorrhiza. It solves the problem of the high cost of using western medicine to control liver cirrhosis and is applicable to the treatment of liver cirrhosis of portal vein type with blood stasis stagnation.

[0005] However, there is no report on a traditional Chinese medicine composition with a moderate number of medicinal flavors and good effects for treating porto-sinusoidal vascular disorder and its application. Summary of the Invention

[0006] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating porto-sinusoidal vascular disorder and its application in view of the deficiencies in the prior art.

[0007] In the first aspect, the present invention provides a traditional Chinese medicine composition for treating porto-sinusoidal vascular disorder, and the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 13 - 17 parts of astragalus membranaceus, 13 - 17 parts of angelica sinensis, and 4 - 8 parts of leech.

[0008] As a preferred example, the traditional Chinese medicine composition is made from the following raw materials by weight: 14-16 parts of Astragalus membranaceus, 14-16 parts of Angelica sinensis, and 5-7 parts of Hirudo.

[0009] As a preferred example, the traditional Chinese medicine composition is made from the following raw materials by weight: 15 parts of Astragalus membranaceus, 15 parts of Angelica sinensis, and 6 parts of Hirudo.

[0010] In a second aspect, the present invention provides the use of the traditional Chinese medicine composition in the preparation of a medicament for treating portal sinusoidal vascular disease of the liver.

[0011] As a preferred example, the liver index of the portal sinusoidal vascular disease of the liver is significantly reduced, and the spleen index is improved.

[0012] As a preferred example, the reticular staining and Sirius red staining of the liver tissue pathology of the portal sinusoidal vascular disease of the liver show: both are relieved.

[0013] The advantages of the present invention are as follows: By feeding SD rats a selenium-rich diet and simultaneously injecting the FOLFOX chemotherapy regimen into the tail vein once a week for 8 consecutive weeks, an animal model of PSVD was replicated. During the modeling process, the modified Danggui Buxue Decoction of the present invention was given for intervention, with simvastatin as the positive control. The results show that the modified Danggui Buxue Decoction can significantly reduce the portal vein pressure of rats and improve the manifestations related to liver microvascular lesions. The liver index is significantly reduced, and the spleen index is improved. The reticular staining and Sirius red staining of the liver tissue pathology show: both are relieved. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 : The technical roadmap of the present invention.

[0015] Figure 2 : Effects of the modified Danggui Buxue Decoction on ALT, AST, liver-body ratio, and spleen-body ratio of PSVD rats.

[0016] Figure 3 : Effects of the modified Danggui Buxue Decoction on the liver tissue pathology of PSVD rats. DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content recorded in the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

[0018] Example 1 Traditional Chinese Medicine Composition (I) for Treating Portal Sinusoidal Vascular Disease of the Liver

[0019] 15 parts of Astragalus membranaceus, 15 parts of Angelica sinensis, and 6 parts of Hirudo.

[0020] Example 2 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (II)

[0021] Astragalus membranaceus 15 parts, Angelica sinensis 13 parts, Hirudo 8 parts.

[0022] Example 3 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (III)

[0023] Astragalus membranaceus 13 parts, Angelica sinensis 17 parts, Hirudo 5 parts.

[0024] Example 4 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (IV)

[0025] Astragalus membranaceus 17 parts, Angelica sinensis 14 parts, Hirudo 7 parts.

[0026] Example 5 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (V)

[0027] Astragalus membranaceus 14 parts, Angelica sinensis 16 parts, Hirudo 6 parts.

[0028] Example 6 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (VI)

[0029] Astragalus membranaceus 16 parts, Angelica sinensis 15 parts, Hirudo 4 parts.

[0030] Example 7 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (VII)

[0031] Astragalus membranaceus 15 parts, Angelica sinensis 17 parts, Hirudo 5 parts.

[0032] Example 8 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (VIII)

[0033] Astragalus membranaceus 13 parts, Angelica sinensis 14 parts, Hirudo 7 parts.

[0034] Example 9 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (IX)

[0035] Astragalus membranaceus 17 parts, Angelica sinensis 16 parts, Hirudo 6 parts.

[0036] Example 10 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (X)

[0037] Astragalus membranaceus 14 parts, Angelica sinensis 15 parts, Hirudo 4 parts.

[0038] Example 11 Traditional Chinese Medicine Composition for Treating Portal Venous Sinus Vascular Disease (XI)

[0039] Astragalus membranaceus 16 parts, Angelica sinensis 13 parts, Hirudo 8 parts.

[0040] Example 12 Animal Experiment

[0041] Study on the Intervention Effect and Mechanism of Modified Danggui Buxue Decoction of the Invention on PSVD Induced by FOLFOX Combined with Selenium-Rich Diet in Rats

[0042] 1. Research overview:

[0043] A PSVD model of rats was induced by FOLFOX combined with selenium-rich diet. Using simvastatin as a control, different doses of modified Danggui Buxue Decoction were given for intervention treatment. By observing the pathology of portal vein blood vessels and hepatic sinusoids, portal vein pressure, etc., the therapeutic effect of modified Danggui Buxue Decoction of the invention on PSVD was explored. The technical roadmap of the invention is shown in Figure 1 .

[0044] 2. Experimental animals:

[0045] 60 clean-grade SD rats at 10 weeks of age, male, with a body weight of 160g ～ 180g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and raised in the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine. They were adaptively raised with free diet and water for 2 weeks. All experiments and operations were carried out in accordance with the ethical requirements of experimental animals of Shanghai University of Traditional Chinese Medicine.

[0046] 3. Drugs and quality control:

[0047] Preparation of aqueous extract of traditional Chinese medicine compound: According to the provisions of the 2020 edition of the Pharmacopoeia of the People's Republic of China, using genuine medicinal materials, the traditional Chinese medicine is from Gansu Province, and the cut crude drugs are purchased from Shanghai Huayu Pharmaceutical Co., Ltd. and identified by experts in pharmacognosy.

[0048] Modified Danggui Buxue Decoction of the invention: Astragalus membranaceus 15g, Angelica sinensis 15g, Hirudo 6g. First, decoct with 6 times the volume of water for 1 hour, then decoct with 8 times the volume of water for 1.5 hours. The decoction liquids of the two decoctions are mixed, filtered, and concentrated into an aqueous extract containing 1.4g / ml of crude drug. The dose conversion of rats for administration is shown in Table 1.

[0049] Table 1 Dose conversion of rats for administration

[0050]

[0051] Simvastatin (SVS): It is an orally active competitive inhibitor of HMG-CoA reductase (HMGCR), which can reduce cholesterol synthesis and lower cholesterol levels in the blood. Simvastatin can significantly reduce the HVPG level of patients, but has no effect on mean arterial pressure, systemic vascular resistance, and hepatic blood flow. It is a newly added vasoprotective agent in Baveno VII, so it is used as the positive control drug in this experiment. The intragastric administration dose of rats is 5mg / kg / d (equivalent to 7 times the dose of a 60kg adult). 4. Model preparation:

[0052] After 2 weeks of adaptive feeding of SD rats, the model group and each drug administration group were given a diet containing 4 ppm Se (rich in selenium), and at the same time, the modeling working solution was injected into the tail vein once a week for 8 consecutive weeks. Preparation method of the modeling working solution: Oxaliplatin (3 mg / kg), 5-fluorouracil (25 mg / kg), and folinic acid (45 mg / kg) were dissolved in 5% glucose solution, and the solution was injected into the tail vein at a dose of 4 ml / kg of the rat body weight. The normal control group was injected with an equal volume of 5% glucose solution into the tail vein.

[0053] 5. Animal grouping and drug administration:

[0054] SD rats were randomly divided into: normal control group (10 rats), PSVD model group (10 rats), model + SVS group (10 rats), model + low- and high-dose Jiawei Danggui Buxue Decoction groups (10 rats each), for a total of 50 rats. After 2 weeks of adaptive feeding, the corresponding drugs were given to each drug intervention group, and the gavage volume was 10 ml / kg of the rat body weight. The normal control group and the model control group were given an equal volume of double-distilled water by gavage.

[0055] 6. Main observation contents and methods:

[0056] (1) Efficacy evaluation:

[0057] Sample collection and preparation: After anesthesia by intraperitoneal injection of 3% sodium pentobarbital, blood was taken from the inferior vena cava. The blood was allowed to stand at 4°C for 2 hours and then centrifuged to collect the serum, which was stored at -70°C. Liver tissue at the same location was excised and fixed in neutral formalin solution, and then dehydrated and paraffin-embedded for pathological examination; the remaining liver tissue was quickly frozen in liquid nitrogen for subsequent detection of protein, nucleic acid, and hydroxyproline content.

[0058] Serum liver function: ALT and AST were measured using kits;

[0059] Paraffin pathology of liver tissue: HE staining was used to observe liver tissue inflammation, Sirius red staining and reticular fiber staining were used to observe collagen deposition in liver tissue, and an automatic digital image system was used to evaluate the inflammation grade and liver fibrosis stage;

[0060] Portal vein pressure: Direct puncture manometry method (open the manometry kit, connect the two ends of the glass tube to the scalp needle and rubber tube respectively. After checking the airtightness of the device, fill the glass tube with 0.9% NaCl solution, remove the air bubbles, and clamp the rubber tube with a vascular clamp. After the rat is anesthetized, open the abdominal cavity, puncture and intubate the main portal vein, release the vascular clamp, and read the value after the water column in the glass tube is stable. Measure 3 times and take the average value);

[0061] (2) Statistical analysis:

[0062] Statistical analysis was performed using SPSS 21.0 software, and graphs were plotted using GraphPad Prism 6 software. Semi-quantitative analysis of image data was performed using fully automated digital image analysis. For measurement data, normality and homogeneity of variance tests were first conducted. If the data conformed to a normal distribution, it was expressed as Mean±SD; if the data did not conform to a normal distribution, it was expressed as Median(Max,Min). If the data conformed to a normal distribution and had homogeneous variance, one-way ANOVA was used for statistical analysis. If there were statistically significant differences between groups, further pairwise comparisons were made using the LSD test; if the data did not conform to a normal distribution or had inhomogeneous variance, the Kruskal-Wallis test was used for statistical analysis. If there were statistically significant differences between groups, further pairwise comparisons were made using the Games-Howell test. Count data was expressed as n(%), and 2 the χ

[0063] 7. Results:

[0064] (1) Effects of the modified Danggui Buxue Decoction on general indicators of PSVD rats

[0065] The effects of the modified Danggui Buxue Decoction of the present invention on ALT, AST, liver-to-body ratio, and spleen-to-body ratio in PSVD rats are shown in Figure 2 . There were no statistically significant differences in ALT and AST among the groups of rats. Only AST showed an increasing trend in the model group and an improving trend in the drug-administered groups. Compared with the normal group, the liver and spleen indices of the model group were significantly increased (P<0.05); compared with the model group, the liver indices of each drug intervention group were significantly decreased (P<0.05), with the high-dose group of the modified Danggui Buxue Decoction being the most obvious. In terms of the spleen index, each drug intervention group showed improvement, and there were statistically significant differences in the high-dose group of the modified Danggui Buxue Decoction and the simvastatin group (P<0.05).

[0066] (2) Effects of the modified Danggui Buxue Decoction on hemodynamics of PSVD rats

[0067] The effects of the modified Danggui Buxue Decoction of the present invention on the hemodynamics of PSVD rats are shown in Table 2. The hemodynamic results showed that compared with the normal group, the portal vein pressure of the model group was significantly increased (P<0.05); compared with the model group of rats, the portal vein pressure of each drug-administered group decreased to some extent, but only the high-dose group of the modified Danggui Buxue Decoction had a statistically significant difference (P<0.05). In addition, there were no statistically significant differences in the mean arterial pressure and heart rate among the groups.

[0068] Table 2 Effects of the modified Danggui Buxue Decoction on the hemodynamics of PSVD rats

[0069]

[0070] Note: * P < 0.05 compared with the normal group; # P < 0.05 compared with the model group.

[0071] (3) Effect of Jiawei Danggui Buxue Decoction on liver tissue pathology of PSVD rats

[0072] The effect of the Jiawei Danggui Buxue Decoction of the present invention on liver tissue pathology of PSVD rats is shown in Figure 3 . HE staining showed that: spotty necrosis, obvious NSD and OPV appeared in the model group, while each administration group was relieved to varying degrees, and the high-dose group of Jiawei Danggui Buxue Decoction and the simvastatin group had significant effects. Reticular staining and Sirius red staining showed that: obvious ISF appeared in the model group, while each administration group was relieved to varying degrees, and the high-dose group of Jiawei Danggui Buxue Decoction and the simvastatin group had significant effects. In addition, the manifestation of NRH was not observed in the model and each administration group.

[0073] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the method of the present invention, several improvements and supplements can be made, and these improvements and supplements should also be regarded as the protection scope of the present invention.

Claims

1. A Chinese medicine composition for treating portal sinusoidal vasculopathy, characterized in that: The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 13-17 parts of astragalus, 13-17 parts of angelica, and 4-8 parts of leech.

2. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 14-16 parts of astragalus, 14-16 parts of angelica, and 5-7 parts of leech.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that: The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 15 parts of astragalus, 15 parts of angelica and 6 parts of leech.

4. Use of the Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a drug for treating portal sinusoidal vascular disease.

5. The use according to claim 4, characterized in that: The liver indexes of the portal sinusoidal vascular disease were significantly reduced and the spleen index was improved.

6. The use according to claim 4, characterized in that: The liver tissue pathological reticular staining and Sirius red staining of the portal sinusoidal vasculopathy showed that all of them were relieved.

Images