基于TLR4炎症通路探讨养胃和络饮对慢性萎缩性胃炎大鼠的作用机制

魏冬梅, 张斌, 王邦才, 王培劼, 何国浓

中国药学杂志 ›› 2023, Vol. 58 ›› Issue (12) : 1102-1109.

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中国药学杂志 ›› 2023, Vol. 58 ›› Issue (12) : 1102-1109. DOI: 10.11669/cpj.2023.12.006
论著

基于TLR4炎症通路探讨养胃和络饮对慢性萎缩性胃炎大鼠的作用机制

  • 魏冬梅a, 张斌a, 王邦才a*, 王培劼a, 何国浓b
作者信息 +

Exploration of the Mechanism of Yangwei-heluo-yin on Rats with Chronic Atrophic Gastritis Based on TLR4 Inflammatory Pathway

  • WEI Dong-meia, ZHANG Bina, WANG Bang-caia*, WANG Pei-jiea, HE Guo-nongb
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文章历史 +

摘要

目的 探究养胃和络饮(YHY)通过抑制Toll 样受体-4(Toll-like receptors-4,TLR4)信号通路改善大鼠慢性萎缩性胃炎(CAG)的作用机制。方法 66只雄性SD大鼠随机分为空白组、模型组、维酶素组、瑞沙托维(TAK242)组、YHY各剂量组,模型组12只,其余每组各9只。除空白组外,其余大鼠采用N-甲基-N'-硝基-N'-亚硝基胍联合破气苦降药、饥饱失常和疲劳多因素复合法构建CAG模型,8周后YHY各剂量组每天分别予以75、150、300 mg·kg-1·d-1 YHY提取液,维酶素组给予维酶素混悬液240 mg·kg-1·d-1,TAK242组给予TAK242溶液1.0 mg·kg-1,空白组和模型组大鼠给予等体积生理盐水,分别灌胃12周。苏木精-伊红(HE)染色法观察大鼠胃黏膜组织病理变化,酶联免疫吸附测定法(ELISA)检测血清和胃黏膜组织白细胞介素(IL)-6、肿瘤坏死因子α(TNF-α)水平,荧光染色定量大鼠胃黏膜组织TLR4、磷酸化丝裂原活化蛋白激酶(p-MAPK)表达,蛋白质免疫印迹法检测胃黏膜组织基质金属蛋白酶2(MMP2)、组织金属蛋白酶抑制剂1(TIMP1)蛋白含量。结果 实验结束后,与空白组相比,模型组大鼠出现胃黏膜层断裂,炎细胞浸润;血清和胃黏膜组织中IL-6、TNF-α水平显著升高(P<0.01);另外,胃黏膜组织TLR4、p-MAPK荧光强度增强;MMP2、TIMP1蛋白含量同时呈现高表达(P<0.01)。与模型组相比,TAK242、维酶素及YHY干预后,胃黏膜组织损伤改善,IL-6、TNF-α水平降低;同时,TLR-4过度表达被抑制,p-MAPK荧光表达减弱;MMP2、TIMP1蛋白含量明显下调(P<0.01);其中以300 mg·kg-1·d-1YHY组改善最为明显。结论 YHY可通过抑制TLR4信号通路,减少炎性因子释放,进而调节胃组织炎症微环境,缓解胃黏膜损伤,从而改善慢性萎缩性胃炎的发展、癌变。

Abstract

OBJECTIVE To study the mechanism of Yangwei-heluo-yin (YHY) in improving chronic atrophic gastritis (CAG) in rats by inhibiting the Toll-like receptor-4 (TLR4) signaling pathway. METHODS Sixty-six male Sprague-Dawley rats were randomly divided into blank group, CAG group, vitacoenzyme group, resatorvid (TAK242) group, YHY treatment groups,with 12 rats in model group and 9 rats in each of the rest groups. Except for the blank group, the other rats were treated with N-methyl-N'-nitro-N'-nitrosoguanidine combined with relieving stagnant Qi and bitter down, hunger and satiety disorders and fatigue synthesis modeling methods to establish CAG model. After 8 weeks, the rats in YHY groups were given with 75, 150, 300 mg·kg-1·d-1 YHY extract every day, the vitacoenzyme group was given with 240 mg·kg-1·d-1 vitacoenzyme suspension, TAK242 group was given with 1.0 mg·kg-1 TAK242 solution, and the blank group and the model group rats were given the same volume of normal saline for 12 weeks by gavage. The histopathological changes of rat gastric mucosa were observed by hematoxylin-eosin (HE) staining. The serum levels of interleu-kin (IL)-6 and tumor necrosis factor α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). The fluorescence expressions of TLR4 and phosphorylated mitogen-activated protein kinase (p-MAPK) in rat gastric mucosa were detected. The expressions of MMP2 and TIMP1 proteins were detected by Western blot. RESULTS At the end of the experiment,compared with the blank group, the gastric mucosa of CAG group was broken and inflammatory cell infiltration appeared, and the levels of IL-6 and TNF-α levels in serum or gastric mucosa tissues of the CAG group increased significantly (P<0.01). In addition, the fluorescence intensities of TLR4 and p-MAPK in tissues were enhanced, and the levels of MMP2 and TIMP1 protein were also highly expressed (P<0.01). Compared with the CAG group, the damage of gastric mucosa tissue was improved after the intervention of TAK242, vitacoenzyme, and YHY, and the levels of IL-6 and TNF-α were decreased. Meanwhile, the over-expression of TLR-4 was inhibited and the fluorescence expression of p-MAPK was decreased. The protein contents of MMP2 and TIMP1 were significantly down-regulated (P<0.01). The improvement was most obvious in 300 mg·kg-1·d-1 YHY group. CONCLUSION YHY can inhibit TLR4 signaling pathway, reduce the release of inflammatory factors, and regulate the inflammatory microenvironment of gastric tissue, relieve gastric mucosal damage, and improve the development of cancer in chronic atrophic gastritis.

关键词

养胃和络饮 / 慢性萎缩性胃炎 / TLR4致炎通路 / 基质金属蛋白酶 / 组织金属蛋白酶抑制剂

Key words

yangwei-heluo-yin / chronic atrophic gastritis / TLR4 inflammatory pathway / matrix metalloproteinase / tissue inhibitor of metalloproteinase

引用本文

导出引用
魏冬梅, 张斌, 王邦才, 王培劼, 何国浓. 基于TLR4炎症通路探讨养胃和络饮对慢性萎缩性胃炎大鼠的作用机制[J]. 中国药学杂志, 2023, 58(12): 1102-1109 https://doi.org/10.11669/cpj.2023.12.006
WEI Dong-mei, ZHANG Bin, WANG Bang-cai, WANG Pei-jie, HE Guo-nong. Exploration of the Mechanism of Yangwei-heluo-yin on Rats with Chronic Atrophic Gastritis Based on TLR4 Inflammatory Pathway[J]. Chinese Pharmaceutical Journal, 2023, 58(12): 1102-1109 https://doi.org/10.11669/cpj.2023.12.006
中图分类号: R965   

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基金

宁波市自然科学基金项目《养胃和络饮对慢性萎缩性胃炎大鼠MMPS/TIMPS表达的影响》资助(2018A610423);宁波市医学重点学科建设项目资助(2022-Z06)
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