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目的:探讨幽门螺杆菌(Helicobacter pylori)对人胃癌MKN45细胞环氧合酶2(Cyclooxygenase-2,COX-2)启动子荧光素酶报告基因重组质粒(pGL3-Basic-COX-2-promoter)活性的影响和健脾解毒方对其的调控作用及可能的机制。方法:将构建的pGL3-Basic-COX-2-promoter转染MKN45细胞,观察H.pylori对其活性的影响。运用p38MAPK特异性抑制剂SB203580阻断p38MAPK信号通路,观察H.pylori对MKN45细胞COX-2启动子活性的影响。Western blot法检测健脾解毒方对H.pylori感染的MKN45细胞p38MAPK信号通路及其下游激活转录因子-2(ATF-2)表达的影响。结果:H.pylori可增加COX-2启动子活性;抑制p38MAPK信号通路后,COX-2启动子活性明显下调;健脾解毒方能够抑制H.pylori诱导的p38MAPK及ATF-)的活性。结论:H.pylori通过p38MAPK信号通路上调胃癌细胞COX-2启动子活性;健脾解毒方通过调控p38MAPK/ATF-2信号通路,抑制COX-2启动子活性,是其防治H.pylori诱发胃癌的机制之一。
Objective: To investigate the effect of Helicobacter pylori on the activity of pGL3-Basic-COX-2-promoter of COX-2 promoter in human gastric cancer MKN45 cells. Effect of Jianpi Jiedu prescription and its possible mechanism. Methods: The constructed pGL3-Basic-COX-2-promoter was transfected into MKN45 cells to observe the effect of H.pylori on its activity. P38MAPK specific inhibitor SB203580 was used to block p38MAPK signaling pathway and the effect of H.pylori on COX-2 promoter activity in MKN45 cells was observed. The effect of Jianpi Jiedu Decoction on the p38 MAPK pathway and the expression of downstream activated transcription factor-2 (ATF-2) in MKN45 cells infected with H.pylori was detected by Western blot. Results: The activity of COX-2 promoter was increased by H.pylori. The activity of COX-2 promoter was down-regulated after p38MAPK signaling pathway was inhibited. Jianpi Jiedu decoction could inhibit the activity of p38MAPK and ATF- induced by H.pylori. Conclusions: H.pylori up-regulates the COX-2 promoter activity in gastric cancer cells via the p38MAPK signaling pathway. Jianpi Jiedu Decoction can inhibit the COX-2 promoter activity by regulating p38MAPK / ATF-2 signaling pathway, One of the mechanisms.