基于NF-κB信号通路探讨萎胃通调汤对大鼠慢性萎缩性胃炎的作用机制

Mechanism of Weiwei Tongtiao Decoction Against Chronic Atrophic Gastritis in Rats:Based on NF-κB Signaling Pathway

目的:探析萎胃通调汤对大鼠慢性萎缩性胃炎(CAG)的干预作用及其可能的机制。方法:90只SD大鼠随机分为正常组,模型组,萎胃通调汤高、中、低剂量干预组,胃复春组6组,每组15只,采用N-甲基-N’-硝基-N-亚硝基胍(MNNG)多因素复合造模法制备CAG大鼠模型。经组织病理学检测鉴定模型制备成功后,给药组分别灌胃给予18、9、4.5 g·kg^(-1)·d^(-1)的萎胃通调汤及0.45 g·kg^(-1)·d^(-1)胃复春水溶液,12周后取大鼠胃黏膜组织经苏木素-伊红(HE)染色后,光学显微镜下观察胃黏膜CAG相关病理学表现;采用实时荧光定量聚合酶链式反应(Real-time PCR)、免疫组织化学法(IHC)及蛋白免疫印迹法(Western blot)分别分析大鼠胃黏膜组织核转录因子-κB(NF-κB)信号通路关键指标mRNA及蛋白表达水平。结果:CAG大鼠胃黏膜固有腺体排列和形态极不规则,伴不同程度减少或消失,固有层可见炎性细胞浸润,约48.4%出现肠上皮化生表现。萎胃通调汤处理后显著改善CAG大鼠胃黏膜固有腺体减少与肠化生等病理改变,呈一定的剂量依赖关系。与正常组比较,模型组大鼠胃黏膜组织环氧合酶2(COX-2)、NF-κB p65、白细胞介素(IL)-1α、IL-1β、IL-10、IL-8α、IL-8βmRNA及COX-2、NF-κB p65、IL-10蛋白表达水平显著上调(P<0.01)。萎胃通调汤及胃复春干预后CAG大鼠胃黏膜组织COX-2、NF-κB p65、IL-1α、IL-1β、IL-10、IL-8α、IL-8βmRNA及COX-2、NF-κB p65、IL-10蛋白表达水平显著下调(P<0.01),其中萎胃通调汤高剂量组相应指标表达水平接近正常组。结论:萎胃通调汤能改善甚至逆转CAG大鼠病变胃黏膜,其作用机制可能与萎胃通调汤下调CAG大鼠胃黏膜组织NF-κB信号通路相关指标mRNA及蛋白表达水平有关。

Objective:To explore the effect of Weiwei Tongtiao decoction(WTD)on chronic atrophic gastritis(CAG)rats and the underlying mechanism.Method:A total of 90 SD rats were randomized into normal control group,model group,high-dose,medium-dose,and low-dose WTD groups(18,9,4.5 g·kg^(-1)·d^(-1)WTD,respectively,ig),and weifuchun control group(0.45 g·kg^(-1)·d^(-1)weifuchun aqueous solution,ig),with 15 rats in each group.The N-methyl-N’-nitro-N-nitrosoguanidine(MNNG)was employed to induced CAG in rats.After the modeling(identified by histopathological examination),the administration began and lasted 12 weeks.Then,gastric mucosa tissues of the rats were collected and stained with hematoxylin and eosin(HE),and the pathological changes of gastric mucosa were observed under the optical microscope.Realtime PCR,immunohistochemistry(IHC),and Western blotting were applied to examine the mRNA and protein expression of indexes in nuclear factor kappa-B(NF-κB)signaling pathway in the gastric mucosa of rats.Result:CAG rats showed irregular arrangement and morphology of the inherent glands in gastric mucosa and the glands decreased or disappeared.In addition,inflammatory cell infiltration was observed in the lamina propria of CAG rats,and about 48.4%presented intestinal metaplasia.WTD significantly alleviated the reduction of the glands and intestinal metaplasia in a dose-dependent manner.Compared with the normal control group,the model group demonstrated increase in the mRNA expression of cyclooxygenase-2(COX-2),NF-κB p65,interleukin(IL)-1α,IL-1β,IL-10,IL-8α,and IL-8β,and protein expression of COX-2,NF-κB p65,and IL-10(P<0.01).WTD and weifuchun lowered the mRNA expression of COX-2,NF-κB p65,IL-1α,IL-1β,IL-10,IL-8α,and IL-8β,and the protein expression of COX-2,NF-κB p65,and IL-10 in gastric mucosa of CAG rats(P<0.01).The expression of the above indexes after the intervention with high-dose WTD was close to that of the normal control group.Conclusion:WTD can improve or even reverse the diseased gastric mucosa of CAG rats,and the mechanism is the likelihood that WTD down-regulates the mRNA and protein expression of the indexes in NF-κB signaling pathway in gastric mucosa of CAG rats.