Application of pinocembrin in preparation of medicine for treating gastric ulcer

Abstract

The invention provides application of pinocembrin in preparation of a medicine for treating gastric ulcer, and belongs to the technical field of medicines. The invention aims to provide the application of the pinocembrin as a unique active ingredient in preparation of the medicine for treating gastric ulcer, and the pinocembrin as the medicine for treating gastric ulcer can reduce the ET-1 content in plasma of rats suffering from gastric ulcer caused by absolute ethyl alcohol, increase the NO content and enhance the repairing effect of gastric mucosa; the contents of TNF-alpha and IL-6 are reduced, and the content of IL-10 is increased; the content of T-SOD in plasma is increased, the content of MPO is reduced, and the antioxidant effect is enhanced. Therefore, the pinocembrin has a remarkable treatment effect on the rat gastric ulcer caused by the absolute ethyl alcohol.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of pine syrup in the preparation of drugs for treating gastric ulcers. Background Technology

[0002] Gastric ulcer (GU) is a common and frequently occurring digestive system disease in clinical practice. It is characterized by a long course, high recurrence rate, numerous complications, and a complex pathogenesis. Currently, it is generally believed that an imbalance between gastric mucosal defense factors (bicarbonate, prostaglandins, and nitric oxide) and aggressive factors (gastric acid and pepsin) is the main cause of this disease. In addition, long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs), excessive alcohol consumption and smoking, excessive mental stress, and Helicobacter pylori infection can also lead to gastric ulcers. Typical symptoms include upper abdominal distension, belching, acid reflux, nausea, stomach pain, decreased appetite, and weight loss. In severe cases, gastric bleeding, ulcer perforation, pyloric obstruction, and even cancer may occur.

[0003] Clinically, commonly used drugs for treating gastric ulcers include proton pump inhibitors (omeprazole, lansoprazole), H2 receptor antagonists (ranitidine, cimetidine), and antacids. While these drugs have advantages such as high specificity and rapid onset of action, they also have limitations and adverse reactions, including high recurrence rates and strong drug resistance. Therefore, finding a gastric ulcer treatment drug with mild side effects and long-lasting efficacy is urgently needed.

[0004] Silver-pine glycoside is a natural trans-stilbene compound, and its structure is shown in the figure below. It is mainly found in plants of the Pinaceae and Lauraceae families. *Lindera reflexa* Hemsl., a plant of the Lauraceae family, is the dried root of the plant. It has a long history of use in traditional medicine for treating gastric ulcers with significant efficacy. It is now clearly established that pine bark extract is the main chemical component of *Lindera reflexa*. Existing technology CN113842390B, "A Composition of Active Ingredients from *Lindera reflexa* with Preventive and Treatment Effects on Gastric Ulcers and its Preparation Method and Application," published on December 28, 2021, describes the treatment of gastric ulcers by extracting a composition of eight active ingredients, including pine bark extract, from *Lindera reflexa*. However, this composition relies on the combined action of all eight active ingredients, resulting in a complex composition and increased medication costs. Furthermore, the inhibition rate of this composition on gastric ulcers is only around 90%, which is relatively low. Therefore, finding a drug that can significantly and effectively treat gastric ulcers is the technical problem this invention aims to solve. Summary of the Invention

[0005] The purpose of this invention is to provide the application of pine syrup as the sole active ingredient in the preparation of a drug for treating gastric ulcers, which has a significant therapeutic effect on gastric ulcers induced by anhydrous ethanol in rats.

[0006] To achieve the above-mentioned invention object, the present invention provides the following technical solutions:

[0007] The present invention provides the use of pinosylvin as the sole active ingredient in the preparation of a drug for treating gastric ulcer, and the structural formula of pinosylvin is:

[0008]

[0009] Preferably, the pinosylvin can reduce the area of gastric ulcer, increase the pain threshold, and decrease the pH value of gastric juice.

[0010] Preferably, the pinosylvin can also improve the blood flow of gastric mucosa, inhibit the inflammatory reaction, inhibit the oxidation reaction, and maintain the integrity of the gastric mucus barrier.

[0011] As a drug for treating gastric ulcer, pinosylvin can decrease the content of ET-1 in the plasma of rats with gastric ulcer induced by absolute ethanol, increase the content of NO, enhance the repair effect of gastric mucosa; and reduce the contents of TNF-α and IL-6, increase the content of IL-10; increase the content of T-SOD in the plasma, decrease the content of MPO, and enhance the antioxidant effect, etc. Therefore, pinosylvin has a significant therapeutic effect on gastric ulcer in rats induced by absolute ethanol. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is the gastric tissue morphology of each group of rats in Example 2. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0013] The technical solutions provided by the present invention will be described in detail below in conjunction with the embodiments, but they cannot be construed as limiting the protection scope of the present invention.

[0014] Experimental Animals

[0015] 20 male healthy SPF-grade SD rats, body weight: 180 g ± 20 g, were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., and the experimental animal license number: SCXK (Lu) 2002-0006. Before the experiment, the rats were raised in an environment with 12-hour day and night alternation, with free access to water and food, and were adaptively fed for 3 days.

[0016] Drugs and Reagents

[0017] Pinus sylvestris was extracted and isolated from *Rhizoma Cylindricae*, and its purity was greater than 98% as analyzed by high performance liquid chromatography. Weitongning tablets were purchased from Henan Lingrui Pharmaceutical Co., Ltd., batch number 2301262. Anhydrous ethanol was purchased from Xinxiang Sanwei Disinfectant Preparation Co., Ltd. Rat ET-1, TNF-α, IL-6, IL-10 enzyme-linked immunosorbent assay (ELISA) kits (catalog numbers: E-HSEL-R0004, E-EL-R0472c, E-EL-R0639c, E-EL-0034c, respectively); T-SOD (WST-1 method), NO, MPO colorimetric assay kits (catalog numbers: E-BC-K020-M, E-BC-K035-M, E-BC-K074-M, respectively) were all from Wuhan Yilairui Biotechnology Co., Ltd.

[0018] Main instruments

[0019] ME203E / 02 electronic balance (Mettler-Toledo Instruments Shanghai Co., Ltd.), PL-200 thermal pain relief device (Chengdu Taimeng Software Co., Ltd.), KZ-Ⅲ-F high-speed tissue grinder (Servicebio), D3024R benchtop high-speed refrigerated centrifuge (Dalong), MX-S vortex mixer (SCILOGEX), HBS-1096A enzyme-linked immunosorbent assay (ELISA) reader (Nanjing Detie), BCD-318 horizontal freezer (Haier), etc.

[0020] Example 1

[0021] Twenty SPF-grade SD rats were randomly divided into four groups of five each: blank control group (KB), model control group (MX), silver pine syrup group (YSS, 45.45 mg / kg), and gastric pain relief tablet group (WT, positive control, 225.00 mg / kg). The YSS and WT groups were treated with the drug by gavage for seven consecutive days, once a day. On the sixth day, the rats were fasted but allowed to drink water at 8 p.m., and the drug was administered 12 hours later. One hour after the drug administration, except for the blank control group, all rats were given an anhydrous ethanol at a dose of 5 ml / kg to create a gastric ulcer model.

[0022] One hour after gavage with anhydrous ethanol, all rats were placed on a thermal pain instrument to measure their pain threshold and record the time it took for them to reach the pain threshold.

[0023] The experimental results of the time to reach the pain threshold in rats by ginsenosides (Table 1) show that the time to reach the pain threshold in the MX group and the WT positive control group were (3.18±0.22)s and (3.92±0.44)s, respectively. Compared with the MX group, the time to reach the pain threshold in the YSS group was significantly increased (P<0.05), which proves that ginsenosides can effectively relieve pain caused by gastric ulcers.

[0024] Table 1 Pain Threshold ( n=5)

[0025]

[0026] Note: Compared with KB, ##P<0.01, indicating a significant difference; compared with MX, *P<0.05, indicating a difference.

[0027] Example 2

[0028] After anesthetizing rats with isoflurane inhalation, the abdomen was incised along the midline to expose the stomach and abdominal aorta. Plasma was collected from the abdominal aorta using a lancet, centrifuged at 5000 g / min for 4 min at 4°C, and the supernatant was collected and stored at -80°C for later use. Next, the pylorus and cardia were ligated with sutures, and a small incision was made at the bottom of the greater curvature of the stomach to collect the gastric contents. The contents were centrifuged at 1000 r / min for 4 min at 4°C, and the supernatant was collected. The gastric fluid volume was recorded and stored at -80°C for later analysis. Finally, the stomach was cut open along the greater curvature, rinsed with physiological saline, unfolded, and photographed.

[0029] 2.1 Observation of gastric tissue morphology and determination of ulcer index

[0030] Observe all the images of gastric tissue taken, and compare the morphology of gastric ulcer edges, ulcer surfaces, and mucosal folds in each group of rats. Use ImageJ software to measure the gastric ulcer index and ulcer inhibition rate under the same conditions.

[0031] Ulcer Index = (Area of ​​Gastric Ulcers) / (Total Area of ​​Stomach) × 100%

[0032] Ulcer inhibition rate = (Ulcer index of model group - Ulcer index of treatment group) / (Ulcer index of model group) × 100%

[0033] The results of stomach tissue imaging are as follows Figure 1 As shown, the gastric mucosa in the KB group was normal in color and smooth in surface, without ulcers or bleeding points; the gastric mucosa in the MX group had a large number of clearly visible hemorrhagic lesions. According to the data in Table 2, the rat gastric ulcer model induced by anhydrous ethanol was successfully established; the YSS group showed only a few punctate hemorrhages visible to the naked eye, which showed a better therapeutic effect compared with the WT group, indicating that ginsenoside has a therapeutic effect on gastric ulcers.

[0034] The effects of ginsenoside on the gastric ulcer index and ulcer inhibition rate in rats are shown in Table 2. Compared with the KB group, the MX group showed a significant difference in the gastric ulcer index (P < 0.01), proving the successful establishment of the anhydrous ethanol-induced gastric ulcer model in rats. The YSS and WT groups also showed significant differences compared with the MX group (P < 0.01), indicating significant therapeutic effects. The ulcer inhibition rate more directly demonstrates the therapeutic effect of ginsenoside.

[0035] Table 2 Gastric ulcer index and ulcer inhibition rate ( n=5)

[0036]

[0037] Note: Compared with KB, ##P<0.01, indicating a significant difference; compared with MX, **P<0.01, indicating a significant difference.

[0038] 2.2 Measurement of gastric juice pH

[0039] Take a portion of the gastric contents supernatant and measure the pH value of the gastric juice using a pH meter.

[0040] Because the gastric fluid volume in the KB group was relatively small, its pH value could not be measured. According to literature review, the normal pH value of gastric fluid in rats is approximately 2.5. Compared with the MX group, the gastric fluid pH value in the YSS group was different (P < 0.05), indicating that ginsenosides have a therapeutic effect on gastric ulcers.

[0041] Table 4. pH values ​​of gastric juice ( n=5)

[0042]

[0043] Note: Compared with the MX group, **P<0.01 indicates a significant difference, and *P<0.01 indicates a significant difference.

[0044] 2.3 Detection of the content of various factors

[0045] Plasma should be used only as needed during testing to avoid repeated freeze-thaw cycles. The levels of ET-1, TNF-α, IL-6, and IL-10 in plasma should be determined using an enzyme-linked immunosorbent assay (ELISA) kit, strictly following the instructions. The levels of T-SOD, MPO, and NO in plasma should be determined using a colorimetric assay kit, also following the instructions.

[0046] 2.3.1 Effect of pine resin on the content of mucosal repair factor in rat plasma

[0047] ET-1 is an active polypeptide with a strong and long-lasting vasoconstrictive effect. This vasoconstrictive effect can lead to decreased blood flow to the gastric mucosa, potentially causing severe ulceration. NO, on the other hand, is an endogenous vasodilator that effectively dilates blood vessels and increases blood flow to the gastric mucosa. When the mucosa is damaged, NO improves gastric mucosal microcirculation, enhancing local blood and oxygen supply, and promoting the renewal and repair of the gastric mucosal epithelium.

[0048] Table 7 shows that, compared with the MX group, the plasma ET-1 levels in both the YSS and WT groups were decreased, and the differences were statistically significant (P < 0.01); the plasma NO levels were increased in both groups. These data indicate that ginseng can reduce plasma ET-1 levels and increase NO levels, thereby increasing gastric mucosal blood flow, promoting gastric mucosal repair, and achieving a therapeutic effect on gastric ulcers.

[0049] Table 7 Changes in plasma ET-1 and NO levels ( n=5)

[0050]

[0051] Note: Compared with KB, ##P<0.01, indicating a significant difference; compared with MX, **P<0.01, indicating a significant difference, and *P<0.05, indicating a significant difference.

[0052] 2.3.2 Effects of pine resin on inflammatory factors in rat plasma

[0053] Gastric ulcers are always accompanied by inflammation. After an inflammatory response occurs, macrophages secrete the pro-inflammatory factor TNF-α, which in turn promotes the production of other inflammatory factors such as IL-6. These factors collectively promote the aggregation of neutrophils at the site of inflammation, exacerbating the severity of the gastric ulcer. IL-10 is a multifunctional negative regulator that inhibits and antagonizes the production and action of pro-inflammatory factors, thereby suppressing the inflammatory response.

[0054] Table 8 shows that the levels of TNF-α, IL-6, and IL-10 in plasma were significantly different between the MX group and the KB group (P < 0.01). Compared with the MX group, the YSS group had lower levels of TNF-α and IL-6 and higher levels of IL-10 in plasma, indicating that ginsenosides can exert anti-inflammatory effects by reducing the levels of TNF-α and IL-6 and increasing the levels of IL-10, thereby treating gastric ulcers.

[0055] Table 8. Plasma levels of TNF-α, IL-6, and IL-10 ( n=5)

[0056]

[0057]

[0058] Note: Compared with KB, ##P<0.01, indicating a significant difference; compared with MX group, **P<0.01, indicating a significant difference, and *P<0.05, indicating a significant difference.

[0059] 2.3.3 Effects of pine resin on antioxidant factors in rat plasma

[0060] Oxidative stress is characterized by elevated levels of intracellular reactive oxygen species (ROS), unpaired molecules that play a crucial physiological role in cellular homeostasis. They damage lipids, proteins, and DNA, leading to lipid peroxidation, cell death, and tissue damage. T-SOD is an important antioxidant widely distributed in organisms; its main function is to resist and prevent damage to cells from oxygen free radicals and to scavenge the oxidation products of oxygen free radicals. MPO is a major component of neutrophil cytoplasmic granules and can promote inflammation, quantitatively reflecting the degree of inflammatory damage.

[0061] As shown in Table 9, compared with the MX group, the plasma T-SOD content in both the YSS and WT groups increased and the MPO content decreased, with significant differences (P < 0.05). This indicates that under the influence of ginsenosides, the plasma T-SOD content in rats increased, the MPO content decreased, and the antioxidant effect was enhanced. Therefore, ginsenosides can exert a therapeutic effect on gastric ulcers through antioxidant activity.

[0062] Table 9. Plasma T-SOD and MPO content ( n=5)

[0063]

[0064] Note: Compared with KB, ##P<0.01, indicating a significant difference; compared with MX, **P<0.01, indicating a significant difference, and *P<0.05, indicating a difference.

[0065] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Use of lariciin as the sole active ingredient in the preparation of a medicament for the treatment of gastric ulcers, characterized in that, The structural formula of the silver pine is:

2. Use according to claim 1, characterized in that, The silver pine can reduce the area of gastric ulcer, increase the pain threshold, and reduce the pH value of gastric juice.

3. Use according to claim 2, characterized in that, The silver pine can also improve the blood flow of gastric mucosa, inhibit inflammatory reaction, inhibit oxidation reaction, and maintain the integrity of gastric mucus barrier.

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