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[合欢皂苷对乙醇诱导的大鼠胃黏膜损伤的保护作用:聚焦于黏液和黏膜屏障]

[Protective effect of Albizia chinensis saponin on ethanol-induced gastric mucosal injury in rats focus on mucus and mucosal barriers].

作者信息

Jia Xiao-Yu, Zhang Jian-Jun

机构信息

State Key Laboratory of Digestive Health, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College Beijing 100050, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2024 Jun;49(12):3340-3347. doi: 10.19540/j.cnki.cjcmm.20240318.701.

Abstract

This study aims to explore the protective effect of Albizia chinensis saponin on ethanol-induced acute gastric ulcer in rats and elucidate its mechanisms. SD rats were deprived of water for 24 hours before the experiment. The control group and model group were administered water by gavage, and the positive drug group received rabeprazole sodium solution(40 mg·kg(-1)) by gavage. The experimental groups were given different doses of Albizia chinensis saponin solution(3, 10, and 30 mg·kg(-1)). After 30 minutes, the control group received 1.5 mL of water by gavage, while the other groups were administered an equal volume of 95% ethanol for modeling. After six hours, the rats were killed by cervical dislocation, and the stomachs were collected. The ulcer area was measured, and the ulcer index was calculated. Hematoxylin-eosin(HE) staining was performed to assess histopathological changes in gastric tissue. Periodic acid-Schiff(PAS) staining was used to evaluate the distribution of gastric mucosal surface mucus. Enzyme-linked immunosorbent assay(ELISA) was employed to measure the levels of phospholipids and aminohexose in the gastric mucosa. Western blot was performed to determine the expression levels of the bicarbonate transporter, matrix metalloproteinase, and tight junction-associated proteins in gastric tissue. Immunohistochemistry(IHC) staining was conducted to quantify the number of positive cells for secreted mucin and tight junction-associated proteins. The results showed that the gastric tissue surface of rats in the control group was smooth without ulceration, and the gastric ulcer index of rats in the model group was 35±11. Albizia chinensis saponin at doses of 3, 10, and 30 mg·kg~(-1) resulted in inhibition rates of gastric ulcer of 46%(P<0.01), 85%(P<0.001), and 100%(P<0.001), respectively. Severe disruption of gastric mucosal structure and absence of the mucus layer were observed in the model group. Compared with the model group, the Albizia chinensis saponin group showed intact gastric mucosal surface mucus layer, significantly increased levels of phospholipids and aminohexose in the mucus, increased number of MUC5AC positive cells, and upregulated expression levels of the bicarbonate transporter SLC26A3 and CFTR. It also showed decreased phosphorylation of JNK and c-Jun, reduced expression levels of MMP-8, elevated expression of TIMP-1, and increased expression levels of Occludin and ZO-1. In conclusion, Albizia chinensis saponin enhances the function of the mucus-bicarbonate barrier by upregulating the content of MUC5AC, phospholipids, and aminohexose and increasing the expression levels of the bicarbonate transporter SLC26A3 and CFTR. Moreover, Albizia chinensis saponin exerts its protective effects on gastric ulcers by inhibiting the JNK signaling pathway to prevent excessive activation of MMP-8, thereby reducing the degradation of Occludin and ZO-1 and enhancing the mucosal barrier function. In summary, Albizia chinensis saponin exerts its anti-gastric ulcer effects by simultaneously enhancing the mucus barrier and the mucosal barrier.

摘要

本研究旨在探讨合欢皂苷对乙醇诱导的大鼠急性胃溃疡的保护作用,并阐明其作用机制。实验前,将SD大鼠禁水24小时。对照组和模型组经口灌胃给予水,阳性药物组经口灌胃给予雷贝拉唑钠溶液(40 mg·kg⁻¹)。实验组给予不同剂量的合欢皂苷溶液(3、10和30 mg·kg⁻¹)。30分钟后,对照组经口灌胃给予1.5 mL水,而其他组给予等体积的95%乙醇进行造模。6小时后,通过颈椎脱臼处死大鼠,采集胃组织。测量溃疡面积,计算溃疡指数。采用苏木精-伊红(HE)染色评估胃组织的组织病理学变化。采用过碘酸-希夫(PAS)染色评估胃黏膜表面黏液的分布。采用酶联免疫吸附测定(ELISA)法检测胃黏膜中磷脂和氨基己糖的水平。采用蛋白质免疫印迹法检测胃组织中碳酸氢盐转运体、基质金属蛋白酶和紧密连接相关蛋白的表达水平。采用免疫组织化学(IHC)染色定量分泌型黏蛋白和紧密连接相关蛋白的阳性细胞数量。结果显示,对照组大鼠胃组织表面光滑无溃疡,模型组大鼠胃溃疡指数为35±11。3、10和30 mg·kg⁻¹剂量的合欢皂苷导致胃溃疡抑制率分别为46%(P<0.01)、85%(P<0.001)和100%(P<0.001)。模型组观察到胃黏膜结构严重破坏且黏液层缺失。与模型组相比,合欢皂苷组胃黏膜表面黏液层完整,黏液中磷脂和氨基己糖水平显著升高,MUC5AC阳性细胞数量增加,碳酸氢盐转运体SLC26A3和CFTR的表达水平上调。还显示JNK和c-Jun的磷酸化降低,MMP-8的表达水平降低,TIMP-1的表达升高,Occludin和ZO-1的表达水平升高。综上所述,合欢皂苷通过上调MUC5AC、磷脂和氨基己糖的含量以及增加碳酸氢盐转运体SLC26A3和CFTR的表达水平来增强黏液-碳酸氢盐屏障的功能。此外,合欢皂苷通过抑制JNK信号通路来防止MMP-8过度活化,从而减少Occludin和ZO-1的降解并增强黏膜屏障功能,对胃溃疡发挥保护作用。总之,合欢皂苷通过同时增强黏液屏障和黏膜屏障发挥抗胃溃疡作用。

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