温阳消饮法对胸腔积液大鼠小肠AQP4及cAMP-PKA信号通路表达的影响

Influence of warming yang for resolving fluid retention therapy on expression of aquaporin-4 and c AMP-PKA pathway in small intestine of rats with pleural effusion

目的:探讨温阳消饮法对胸腔积液模型大鼠小肠水通道蛋白4(A Q P4)的表达、环磷酸腺苷(c AMP)、蛋白激酶A(PKA)含量的影响,阐释该法的作用机制。方法:用计算机随机分组方法,将Wistar成年大鼠随机分为温阳消饮组、去桂消饮组、模型对照组、空白对照组,每组10只。除K组外,其余3组动物以1%角叉菜胶胸腔注射建立大鼠胸腔积液(悬饮)模型,造模24h后拍摄X线胸片进行模型评价。于造模成功后2h各组予相应药物灌胃,每天1次,连续5d。灌胃结束后处死各组大鼠,采取小肠组织,用蛋白印迹法检测AQP4,用酶联免疫吸附法检测c AMP、PKA。结果:1AQP4表达:与空白对照组比较,模型对照组显著降低(P<0.05);与模型对照组比较,温阳消饮组、去桂消饮组均显著增高(P<0.05);2c AMP含量:各组比较,其差异均无统计学意义;3PKA含量:与K组比较,其余各组均显著升高(P<0.05);与模型对照组比较,温阳消饮组、去桂消饮组的差异无统计学意义。结论:在胸腔积液模型中,大鼠小肠上段AQP4的表达降低,而温阳消饮法可上调之,这可能与该法治疗水饮病的作用机制有关。

Objective： To explore the influence of warming yang for resolving fluid retention therapy on expression of aquaporin-4（AQP4）, cyclic adenosine monophosphate（c AMP） and protein kinase A（PKA） in small intestine of rats with pleural effusion, and to reveal its possible mechanism. Methods： Using the digitally random methods to divide the adult Wistar rats into four groups： warming yang for resolving fluid retention group（W）, warming yang for resolving fluid retention without cassia twig group（Q）, model control group（M） and bank control group（K）, each group with 10 rats. All rats expect those in K group received intrapleural injection of 1% carrageenan to establish the rat model of pleural effusion（suspending fluid retention）. After 24 h, the rat model was evaluated by radiography. 2h after modeling, each group received respective treatments by gavage once daily for 5d. After treatment, the small intestine of rats was collected. The expression of AQP4 was detected by Western blot, the content of c AMP and PKA was determined by enzme-linked immunosorbent assay（ELISA）. Results： 1AQP4： Compared with the K group, the expression of AQP4 in M group was decreased; compared with the M group, the W group and Q group was increased; there was a significant difference between them（P〈0.05）. 3c AMP： The content of c AMP in each group was no significant difference. 2PKA： Compared with the K group, the expression of PKA in other groups were significance increased（P〈0.05）, and other group has no difference. Conclusion： In the rat model of pleural effusion, the expression of AQP4 in empty intestine was reduced, and it could be up-regulated by the therapy of warming yang for resolving fluid retention, this might be one of the mechanisms of warming yang for resolving fluid retention therapy in treating the disease of retained fluid.