Application of Laetanine in preparation of medicine for preventing and / or treating gastric ulcer

Abstract

The invention provides application of Laetanine in preparation of a medicine for preventing and / or treating gastric ulcer, and belongs to the technical field of biological medicine. The medicine prepared from the Laetanine has the effects of relieving gastric ulcer pain, reducing the pH value of gastric juice and reducing the ET-1 content in plasma at the same time. Research confirms that the Lapanine can enable the pH value of gastric juice of a rat to tend to a normal value, increase the contents of PGE2 and T-SOD in plasma and reduce the contents of ET-1, TNF-alpha and IL-6 in the plasma, that is, the Lapanine has the effects of relieving gastric ulcer pain, reducing the pH value of the gastric juice and reducing the content of ET-1 in the plasma at the same time, the specific pharmacological action of the Lapanine in gastric ulcer is further confirmed, and a foundation is laid for clinical use.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the use of Laetanine in the preparation of drugs for the prevention and / or treatment of gastric ulcers. Background Technology

[0002] Gastric ulcer (GU) is a common digestive system disease with a complex pathogenesis. Currently, it is believed to be mainly caused by an imbalance between defensive factors (bicarbonate, prostaglandins, and nitric oxide) and aggressive factors (gastric acid and pepsin) in the gastric mucosa. In addition, long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs), excessive alcohol consumption and smoking, excessive 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 perforation or gastric bleeding may occur.

[0003] Clinically, commonly used drugs for treating gastric ulcers include proton pump inhibitors (omeprazole, lansoprazole), H2 receptor antagonists (ranitidine, cimetidine), and antacids. However, drugs such as omeprazole and lansoprazole either inhibit gastric acid secretion, relieve pain, or increase serum ET-1 levels. There are few drugs that can effectively relieve gastric ulcer pain, lower gastric pH, and reduce plasma ET-1 levels. Furthermore, no research has proven which specific active ingredient can achieve the corresponding effect.

[0004] Laetanine is an alkaloid extracted from the dried root of *Lindera arvensis* Hemsl., a plant belonging to the genus *Lindera* of the family Lauraceae. Commonly known as *Tu Chen Xiang* or *Xiang Gun*, it is mainly produced in the Dabie Mountains of Xinyang, Henan Province. It is believed to have the effects of regulating qi and relieving pain, strengthening the stomach and eliminating food stagnation, stopping bleeding and reducing swelling. It is mainly used to treat stomach distension and pain, stomach cold pain, belching and acid reflux, and peptic ulcers. Current cell studies have shown that the *Lindera arvensis* extract, Laetanine, has an ameliorative effect on acetic acid-induced cell damage, but whether it can treat gastric ulcers or simultaneously relieve gastric ulcer pain, lower gastric pH, and reduce plasma ET-1 levels remains uncertain. Summary of the Invention

[0005] The purpose of this invention is to provide the use of Laetanine in the preparation of medicaments for the prevention and / or treatment of gastric ulcers.

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

[0007] This invention provides the use of Laetanine in the preparation of a drug for the prevention and / or treatment of gastric ulcers, the drug having the effects of relieving gastric ulcer pain, reducing gastric juice pH and reducing plasma ET-1 levels.

[0008] Preferably, the structure of the Laetanine is as shown in formula (1):

[0009]

[0010] (1).

[0011] The present invention has the following beneficial effects:

[0012] This invention provides the application of Laetanine in the preparation of drugs for the prevention and / or treatment of gastric ulcers. Studies have confirmed that Laetanine can normalize the pH value of gastric juice in rats, increase the content of PGE2 and T-SOD in plasma, and decrease the content of ET-1, TNF-α, and IL-6 in plasma. This indicates that Laetanine can achieve its anti-gastric ulcer effect by inducing endogenous molecules to protect cells, improve gastric mucosal blood flow, inhibit inflammatory response, inhibit oxidative response, and maintain the integrity of the gastric mucus barrier.

[0013] Furthermore, it was also shown that Laetanine simultaneously relieves gastric ulcer pain, lowers gastric pH, and reduces plasma ET-1 levels, confirming the specific pharmacological effects of Laetanine extract in gastric ulcers and laying the foundation for clinical use. Attached Figure Description

[0014] Figure 1 This is a morphological diagram of the stomach tissue in Example 3. Detailed Implementation

[0015] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0016] The Laetanine used in the following examples was extracted and isolated from *Cassia tora*, and the extraction and separation method was not limited. High-performance liquid chromatography (HPLC) analysis was performed, and the purity was ≥97%. The Weitongning tablets used were purchased from Henan Lingrui Pharmaceutical Co., Ltd., batch number 2301262. The rat PGE2, ET-1, TNF-α, and IL-6 enzyme-linked immunosorbent assay (ELISA) kits used were purchased from Wuhan Yilairui Biotechnology Co., Ltd., catalog numbers E-EL-R0792c, E-HSEL-R0004, E-EL-R0472c, and E-EL-R0639c, respectively. The T-SOD (WST-1 method) colorimetric assay kit used was purchased from Wuhan Yilairui Biotechnology Co., Ltd., catalog number E-BC-K020-M.

[0017] Example 1: Animal grouping and administration

[0018] Twenty male SPF-grade SD rats, weighing 180 g ± 20 g, were purchased from Jinan Pengyue Laboratory Animal Breeding Co., Ltd., with the experimental animal license number: SCXK(Shandong)2002-0006. Before the experiment, the rats were raised in an environment with a 12-hour day-night cycle, allowed free access to water and food, and adaptively fed for 3 days.

[0019] Twenty SPF-grade SD rats were randomly divided into 4 groups of 5 rats each, namely the blank control group (KB), the model control group (MX), the Laetanine group (LAE), and the Weitengning Tablet group (WT, positive control); the LAE and WT groups were continuously intragastrically administered for 7 days for intervention, once a day. At 8 pm on the sixth day, the rats were fasted but allowed to drink water. After 12 hours, the drugs were administered. One hour after drug administration, except for the blank control group, all rats were intragastrically administered absolute ethanol at a dose of 5 mL / kg to establish a gastric ulcer model.

[0020] Example 2 Determination of the latency of the heat-induced foot withdrawal reflex in rats

[0021] One hour after intragastric administration of absolute ethanol, the rats were placed in an acrylic cage, and the central part of the plantar surface of the rats was irradiated with a PL-200 thermal pain stimulator. The parameters were set as an intensity of 35%, and each continuous photothermal stimulation did not exceed 30 s. When the rats showed rapid foot lifting and foot licking responses, the latency value of the foot withdrawal reflex displayed by the instrument was recorded. It was continuously measured 5 times, and the average of the 5 measurements was taken as the latency of the heat-induced foot withdrawal reflex of the rat.

[0022] Table 1 Latency of the heat-induced foot withdrawal reflex

[0023] Group Dosage (mg / kg) The latency (s) of the heat-shrink foot reflex. KB 0.00 5.14±0.90 MX 0.00 <![CDATA[3.18±0.22 ## <!-- 2 -->]]> LAE 28.08 <![CDATA[4.14±0.70 ** ]]> WT 225.00 <![CDATA[3.92±0.44 * ]]>

[0024] Note: Compared with KB, ##P < 0.01, indicating a significant difference; compared with the MX group, **P < 0.01 indicating a significant difference, and *P < 0.05 indicating a difference.

[0025] As can be seen from Table 1, the latencies of the heat-induced foot withdrawal reflex in the MX group and the WT positive control group were (3.18 ± \alpha)s and (3.92 ± 0.44)s respectively. Compared with the MX group, the latency of the heat-induced foot withdrawal reflex in the LAE group was significantly increased (P < 0.01), and it was longer than that in the WT group, proving that Laetanine can effectively relieve the pain caused by gastric ulcer.

[0026] Example 3 Observation of gastric tissue morphology

[0027] (In the above translation, the symbol \alpha in the text of item should be the correct value in the original Chinese text, but it is not clearly given in the provided content. Here it is retained as \alpha for reference.)After isoflurane inhalation anesthesia, rats were incised along the midline of the abdomen to expose the stomach and abdominal aorta. Plasma was collected from the abdominal aorta using a lancet. 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 stomach was then cut open along the greater curvature, rinsed with physiological saline, unfolded, and photographed. Images of the gastric tissue were observed, and the morphology of bleeding points, gastric ulcer edges, ulcer surfaces, and mucosal folds were compared among the groups of rats. The gastric ulcer index and ulcer inhibition rate were measured using ImageJ software under the same conditions.

[0028] Ulcer Index = (Area of ​​Gastric Ulcers) / (Total Area of ​​Stomach) × 100%

[0029] Ulcer inhibition rate = (Ulcer index of model group - Ulcer index of treatment group) / (Ulcer index of model group) × 100%

[0030] Depend on Figure 1 It can be seen that 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; the WT group showed a large number of linear and punctate hemorrhages under gross observation; while the LAE group had only a few linear hemorrhages and very few punctate hemorrhages, indicating that Laetanine can reduce gastric mucosal bleeding.

[0031] Table 2 Gastric ulcer index and ulcer inhibition rate

[0032] Group Dosage (mg / kg) Ulcer index (%) Inhibition rate (%) KB 0.00 — — MX 0.00 <![CDATA[5.08±0.80 ## ]]> — LAE 28.08 0.74±0.53** 85.43 WT 225.00 1.78±1.51** 64.96

[0033] Note: Compared with KB, ##P<0.01, indicating a significant difference; compared with MX, **P<0.01, indicating a significant difference.

[0034] Table 2 shows that the gastric ulcer index of the MX group was significantly different from that of the KB group (P < 0.01), proving the successful establishment of the anhydrous ethanol-induced gastric ulcer model in rats. The LAE group showed significant differences compared to the MX group (P < 0.01), indicating significant therapeutic effects in both groups. The gastric ulcer index of the LAE group was much smaller than that of the WT group, and the ulcer inhibition rate of the LAE group was 31.51% higher than that of the WT group, indicating that the therapeutic effect of the alkaloid LAE was better than that of the stomach-soothing tablets.

[0035] Example 4: Measurement of gastric juice pH

[0036] Take the gastric contents collected in Example 3, centrifuge at 1000 r / min for 4 min at 4°C, take the supernatant, record the gastric fluid volume, and measure the pH value of the gastric fluid using a pH meter.

[0037] Table 3 pH values ​​of gastric juice

[0038] Group Dosage (mg / kg) gastric juice pH KB 0.00 The gastric fluid volume was small, and the pH value could not be measured. MX 0.00 5.77±1.13 LAE 28.08 4.56±0.38** WT 225.00 4.47±0.69**

[0039] Note: Compared with the MX group, **P < 0.01 indicates a significant difference.

[0040] As shown in Table 3, the gastric juice pH of the LAE group and the WT group was significantly lower than that of the MX group. According to the literature, the normal gastric juice pH of rats is about 2.5. This means that the LAE group can effectively restore the acid secretion function of the gastric mucosa and has a certain therapeutic effect on gastric ulcers caused by anhydrous ethanol.

[0041] Example 5: Detection of the levels of PGE2, ET-1, TNF-α, IL-6, and T-SOD

[0042] Plasma collected from the abdominal artery in Example 3 was centrifuged at 5000g / min for 4 min at 4°C. The supernatant was collected, and the contents of ET-1, TNF-α, IL-6, and PGE2 in the plasma were determined using an enzyme-linked immunosorbent assay (ELISA) kit, strictly following the instructions. The contents of T-SOD in the plasma were determined using a colorimetric assay kit, also following the instructions.

[0043] SIMCA 14.0 software was used to perform PCA (principal component analysis) on the data. SPSS statistical software was used to analyze the experimental results, and the experimental data are expressed as mean ± standard deviation.

[0044] Table 4. Plasma levels of PGE2, ET-1, TNF-α, IL-6, and T-SOD

[0045]

[0046]

[0047] 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.

[0048] As shown in Table 4, compared with the MX group, the plasma PGE2 content in both the LAE and WT groups was increased. The content change in the LAE group was significantly different (P<0.01). This will increase the protective factors on the gastric mucosa, promote mucosal blood circulation and protein synthesis, dilate blood vessels, increase gastric mucosal blood flow, reduce gastric mucosal damage, and have a therapeutic effect on gastric ulcers in rats.

[0049] Compared with the MX group, the plasma ET-1 levels in both the LAE and WT groups were significantly lower (P < 0.01), with the LAE group showing a greater decrease in ET-1 levels. This indicates that LAE can increase gastric mucosal blood flow and promote gastric mucosal repair by reducing plasma ET-1 levels, thereby achieving the therapeutic effect on gastric ulcers.

[0050] Compared with the KB group, the plasma levels of TNF-α and IL-6 in the MX group were significantly different (P < 0.01). The plasma levels of TNF-α and IL-6 in the LAE group and the WT positive control group were lower than those in the MX group, showing a significant difference (P < 0.01). The decrease in TNF-α and IL-6 levels was more pronounced in the LAE group, indicating that Laetanine can exert its anti-inflammatory effect by reducing TNF-α and IL-6 levels, thereby treating gastric ulcers.

[0051] Compared with the MX group, the plasma T-SOD level in the LAE group was significantly higher (P < 0.01), indicating that under the action of Laetanine, the plasma T-SOD level in rats increased, antioxidant factors increased, and antioxidant effect was enhanced. That is, Laetanine exerts its therapeutic effect on gastric ulcers through antioxidant activity.

[0052] In summary, compared with the MX group, Laetanine decreased the pH of gastric juice in rats, bringing it closer to normal; increased plasma PGE2 levels, resulting in stronger gastric mucosal protection; decreased plasma ET-1 levels, enhancing gastric mucosal repair; reduced TNF-α and IL-6 levels, enhancing anti-inflammatory effects; and increased plasma T-SOD levels, enhancing antioxidant effects. This indicates that Laetanine exerts its anti-gastric ulcer effect by inducing endogenous molecules to protect cells, improving gastric mucosal blood flow, inhibiting inflammatory responses, suppressing oxidative reactions, and maintaining the integrity of the gastric mucus barrier. In other words, Laetanine simultaneously relieves gastric ulcer pain, lowers gastric juice pH, and reduces plasma ET-1 levels.

[0053] 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. The use of Laetanine in the preparation of drugs for the prevention and / or treatment of gastric ulcers, characterized in that, The drug has the effects of relieving gastric ulcer pain, lowering gastric pH, and reducing the ET-1 content in plasma.

2. The use of Laetanine according to claim 1 in the preparation of drugs for the prevention and / or treatment of gastric ulcers, characterized in that, The structure of the Laetanine is shown in equation (1):

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