中药萎胃康颗粒剂逆转慢性萎缩性胃炎基于NF-kB相关信号通路的机制研究

Generation mechanism and reversing mechanism of chronic atrophic gastritis are hot spots and difficult problems in the study of gastric disease. It has vital clinical and theoretical significance to prove the pathologic mechanism in the development process of atrophic gastritis, to summary the general rules of generation and reversal of atrophic gastritis and to seek effective methods and drug of reversion,prevention and treatment of atrophic gastritis. This project wil use Wistar rats to make experimental animal model of chronic atrophic gastritis by many kinds of stimulation ,such as primarily filling the stomach using the sodium salicylate.Carry on the reversal treatment with oral administration of Wei-Wei kang granule with different dosage.To explore the effection on the content of the Inflammatory cell factor IL-6 and IL-10, the gamma gene expression of ITF and PPAR mRNA in gastric mucosa and the TLRs/NF-kB pathway from the angle of immune regulation. To explore the effection on the apoptosis of gastric mucosa, the protein expression of p53 and Bcl-2, the gamma gene expression of ITF and the pathway of NF-κB(P65)/CyclinE from the angle promoting gastric mucosal to repair by RT-PCR technology, Western Blotting method , ELASA method and so on.To reveal its inner mechanism and key targets for the treatment of the disease and to study on therapeutic effect of different doses of Weiweikang granule of Chinese traditional medicine. to provide clear dosage instructions for clinic.

慢性萎缩性胃炎的产生机理及其逆转机制是胃病研究中的热点和难点问题，探明萎缩性胃炎产生发展变化过程的病理机制，总结萎缩性胃炎产生和逆转的一般规律，寻求逆转萎缩性胃炎的有效方法和药物对萎缩性胃炎的防治，具有重要的临床价值和理论意义。本项目拟应用Wistar大鼠以水杨酸钠灌胃为主等多重刺激制备萎缩性胃炎动物模型，并服不同剂量的中药萎胃康进行逆转。拟采用RT-PCR技术、Western Blotting及ELASA等方法进行指标检测，从免疫调节角度探讨其对炎性细胞因子IL-6，IL-10的含量、胃粘膜中PPARγmRNA的表达及TLRs/NF-κB通路的影响，从促进胃黏膜修复角度，探讨其对胃黏膜细胞凋亡、p53 和Bcl-2蛋白表达、ITF mRNA表达及NF-κB(P65)/CyclinE通路的影响，从而揭示其治疗该病的内在机制和关键靶点，并探讨中药萎胃康中药萎胃康颗粒不同剂量疗效，以服务临床。

慢性萎缩性胃炎（Chronic atrophic gastritis，CAG）是临床常见病、多发病，且有一定的癌变倾向。本病的病因和发病机理尚未明确。目前西医尚缺乏有效的治疗药物。而中药具有治疗与调理双层作用、毒副作用少等优势。但目前国内外中药治疗CAG的研究尚处于起步阶段。本项目应用SD大鼠以水杨酸钠灌胃为主等多重刺激制备CAG动物模型，并服不同剂量的中药萎胃康进行逆转。用药30天后，观察各组大鼠胃黏膜组织形态的变化。采用ELISA 法检测大鼠血清炎性细胞因子IL-6，IL-10 的含量；免疫组化法和Western Blotting 法检测大鼠胃黏膜NF-kB、Bax、Bcl-2、CyclinE、p53蛋白表达；TUNEL 法检测大鼠胃黏膜细胞凋亡；RT-PCR 技术检测TFF3、PPARγ1、PPARγ2、TLR2、TLR4 mRNA 表达。结果显示各治疗组大鼠胃黏膜病理明显改善，以萎胃康大剂量组效果最好，接近正常状态。与正常组比较，模型组大鼠血清IL-6含量增加，IL-10 含量减少；模型组大鼠胃黏膜凋亡细胞数增加，NF-kB、Bcl-2、CyclinE蛋白表达升高，Bax、p53蛋白表达降低，TFF3、PPARγ1、PPARγ2、TLR2、TLR4 mRNA表达升高。经中西药治疗后，与模型组比较，各治疗组大鼠血清IL-6含量降低，IL-10 含量增加；各治疗组大鼠胃黏膜凋亡细胞数减少，NF-kB、Bcl-2、CyclinE蛋白表达降低，Bax、p53蛋白表达升高，TFF3、PPARγ1、PPARγ2、TLR2、TLR4 mRNA表达降低。其中以萎胃康大剂量组变化最为明显，具有统计学意义。说明中药萎胃康治疗CAG疗效确切，其机制可能与调节免疫与促进胃黏膜修复有关。该研究揭示了中药萎胃康治疗CAG的内在机制和关键靶点，并探讨了中药萎胃康不同剂量疗效，可更好地服务临床。