TGF-MAPK/ERK-c-myc 通路介导慢性萎缩性胃炎癌变机制及加味七方胃痛颗粒干预效应

Cells proliferation and apoptosis in the gastric mucosa in chronic atrophic gastritis(CAG) induced by TGF-MAPK/ERK-c-myc signaling pathway. The abnormal regulations at the different segments of the signaling pathway lead to the accessive cells proliferation and inadequate apoptosis hence to cancerization..Based on the previous clinical and animals experiments,this project is to duplicate CAG rats models by feeding the carcinogen MNNG; to detect the key index of TGF-MAPK/ERK-c-myc pathway by means of mucosa immuohistochemistry some modern molecular biological techniques such as RT-PCR，Western Blot; to observe the changes of TGF-β1/TGF-β1R、Ras、Raf、MEK、ERK，apoptosis GAP c-myc、bcl-2、p53 and proliferation GAP cyclinD1、CDKs etc; to study the cancerization mechanism of CAG induced by TGF-MAPK/ERK-c-myc signaling pathway; to adopt the compound Chinese patent medicine Modified Qifangweitong Granules which has been conformed in effects clinically in the intervention of CAG model rats and to observe it's effects so as to provide experimental basis for the prevention and treatmnet of gastric cancer by Modified Qifangweitong Granules.

TGF-MAPK/ERK-c-myc通路介导慢性萎缩性胃炎（CAG）胃黏膜细胞的增殖与凋亡，在其通路的不同环节出现调控失常，导致胃黏膜细胞过度增殖和凋亡不足，最终导致癌变。本课题在前期临床和实验研究的基础上，以饮用化学致癌剂MNNG复制CAG大鼠模型，运用免疫组化、RT-PCR，Western Blot等现代分子生物学技术检测TGF-MAPK/ERK -c-myc通路的关键指标：TGF-β1/TGF-β1R、Ras、Raf、MEK、ERK，细胞凋亡相关蛋白c-myc、bcl-2、p53及细胞增殖相关蛋白cyclinD1、CDKs等的变化，探讨TGF-MAPK/ERK-c-my通路介导的CAG癌变机制，并运用临床疗效肯定的中药复方"加味七方胃痛颗粒"干预CAG模型大鼠，观察"加味七方胃痛颗粒"对TGF-MAPK/ERK-c-my通路的干预效应，为中药"加味七方胃痛颗粒"防治胃癌提供实验依据。

我国胃癌的发病率及死亡率均仅次于位于恶性肿瘤首位的肺癌，所以对胃癌癌前病变的研究现已成为胃癌二级预防的重要内容。本项目的研究目地是为了探讨TGF- MAPK/ERK-c-myc通路介导胃癌癌前病变（PLGC）及其癌变的病理生理机制，和加味七方胃痛颗粒对TGF- MAPK/ERK-c-myc通路的干预效应。方法：采用饮用化学致癌物MNNG建立胃癌癌前病变大鼠模型，除空白组、模型组外分别给予三种不同剂量加味七方胃痛颗粒和胃复春干预，采用RT-PCR、免疫组化、Western blot检测TGF- MAPK/ERK-c-myc通路各基因的表达水平。结果：1.与模型组对比，加味七方胃痛颗粒及胃复春有效改善 PLGC大鼠模型胃粘膜萎缩、炎症浸润程度、不完全肠上皮化生及异型增生 (P＜0.05)。2.加味七方胃痛颗粒大、中、小组及胃复春组均能降低TGF-β1/TGF-β1-R、Raf、MEK、ERK、c-myc、bcl-2、P53、CyclinD1、CDKs基因的表达，并且明显低于模型组（p﹤0.01）。3.加味七方胃痛颗粒大、中、小组及胃复春组均能有效抑制PLGC模型大鼠TGF-β1/TGF-β1-R、Raf、MEK、ERK、c-myc、bcl-2、P53、CyclinD1、CDKs基因蛋白表达，并且明显低于模型组（p﹤0.01）。结论：1.TGF- MAPK/ERK-c-myc通路各基因的过度表达是PLGC发生发展的机制之一，当TGF- MAPK/ERK-c-myc通路各基因蛋白表达过高时可造成细胞损伤、凋亡及增殖失控。2.加味七方胃痛颗粒具有保护、修复PLGC大鼠模型的胃粘膜。3.TGF- MAPK/ERK-c-myc通路通过下调TGF-β1/TGF-β1-R、Raf、MEK、ERK、c-myc、bcl-2、P53、CyclinD1、CDKs基因及其蛋白的表达来修复损伤细胞及逆转癌前病变。