左归降糖益肾方含药血浆对高糖培养小鼠足细胞podocin及nephrin表达的影响

Effects of Zuogui Jiangtang Yishen decoction plasma on Podocin and Nephrin expressions in podocyte cells of high glucose condition in vitro

关键词:  左归降糖益肾方  高糖  足细胞  Podocin  Nephrin

Key words:  Zuogui Jiangtang Yishen decoction plasma  high glucose  podocyte  Podocin  Nephrin

摘要:

[目的] 探讨在高糖作用下,体外培养的小鼠足细胞podocin及nephrin表达水平的变化,及左归降糖益肾方含药血浆的调控作用。[方法] 将体外培养的小鼠足细胞随机分为空白组(A组)、高糖组(B组)、左归降糖益肾方含药血浆组(C组)、4-苯基丁酸含药血浆组(D组).采用间接免疫荧光细胞化学法鉴定足细胞podocin及nephrin蛋白的表达;噻唑蓝(MTT)自动比色法检测足细胞活力,蛋白质印迹法,逆转录-聚合酶链反应法分别检测podocin及nephrin蛋白或mRNA表达水平的变化。[结果] 与A组相比,B组足细胞活力,podocin及nephrin蛋白或mRNA的表达水平均降低(P <0.01);与B组相比,C组、D组足细胞活力,podocin及nephrin蛋白或mRNA的表达水平均升高(P <0.01);与C组相比,D组足细胞活力升高(P<0.05).[结论] podocin及nephrin蛋白或mRNA表达水平的降低,是足细胞损伤的分子机制之一;左归降糖益肾方含药血浆可以通过升高podocin及nephrin蛋白或mRNA的表达水平,从而保护足细胞。

Abstract:

[Objective] To investigate the levels of protein and mRNA expressions of podocin and nephrin in podocyte cells cultured in high glucose condition in vitro, and the effects of Zuogui Jiangtang Yishen decoction plasma on podocin and nephrin expressions in podocyte cells. [Methods] Cultured podocyte cells were divided into four groups:control group(A group), high glucose group(B group), Zuogui Jiangtang Yishen decoction plasma group(C group) and 4-Phenylbutyric acid group (D group). After incubation for 48 h, the cell viability was detected by MTT colorimetry assay. The protein and mRNA expressions of Podocin and Nephrin were analyzed by western blot or reverse transcription-polymerase chain reaction respectively. [Results] Induced by high g1ucose, the viability of podocyte cells was significantly decreased of B group, and their expressions of Podocin and Nephrin protein or mRNA were downregulated, as compared with A group (P <0.01); as compared with B group, the expressions of Podocin and Nephrin protein or mRNA of podocyte cells of C group and D group were upregulated, and the viability of podocyte cells were significantly increased(P <0.01); and compared with C group, the cell viability was significantly increased of D group (P<0.05). [Conclusion] The lower level expressions of Podocin and Nephrin protein or mRNA were the molecular mechanisms of podocyte injury. ZJYD plasma can repair the damaged podocyte by upregulating Podocin and Nephrin protein or mRNA expressions.