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目的:探讨大黄总蒽醌含药血清对体外培养NRK细胞水通道蛋白2(AQP2)、水通道蛋白4(AQP4)表达的调节效应。方法:16只SD大鼠随机分为正常组、大黄总蒽醌低、中、高剂量组(0,1400,2500,4500mg·kg-1.d-1灌胃),7d后麻醉处死取血制备含药血清。体外培养NRK细胞并给予不同浓度大黄含药血清培养液24h处理,采用免疫荧光检测AQP2、AQP4的定位,Western blot及半定量RT-PCR检测AQP2、AQP4蛋白及mRNA的表达。结果:AQP2、AQP4主要表达在NRK细胞膜上,2500,4500mg·kg-1.d-1大黄总蒽醌含药血清培养液可抑制NRK细胞的AQP2、AQP4蛋白及mRNA的表达,与对照组相比差异有统计学意义(P<0.05)。结论:大黄总蒽醌含药血清可抑制NRK细胞AQP2、AQP4基因转录与翻译,提示大黄的利尿作用可能与调节AQP2、AQP4表达有关。
Objective: To investigate the regulatory effect of rhubarb drug-containing serum on the expression of aquaporin 2 (AQP2) and aquaporin 4 (AQP4) in cultured NRK cells. METHODS: Sixteen SD rats were randomly divided into normal group, low-dose, medium-dose, high-dose group of rhubarb (0, 1400, 2500, 4500 mg·kg-1.d-1 gavage), and blood was taken by anesthesia 7 days later. Preparation of drug-containing serum. NRK cells were cultured in vitro and treated with different concentrations of rhubarb serum-containing medium for 24 h. Immunofluorescence was used to detect the localization of AQP2 and AQP4. Western blot and semi-quantitative RT-PCR were used to detect the expression of AQP2 and AQP4 protein and mRNA. Results: AQP2 and AQP4 were mainly expressed on the membrane of NRK. 2500, 4500 mg·kg-1.d-1 rhubarb serum-containing medium could inhibit the expression of AQP2, AQP4 protein and mRNA in NRK cells, and compared with the control group. The difference was statistically significant (P<0.05). Conclusion: The rhubarb-containing serum can inhibit the transcription and translation of AQP2 and AQP4 genes in NRK cells, suggesting that the diuretic effect of rhubarb may be related to the regulation of AQP2 and AQP4 expression.