左归降糖益肾方含药血浆对高糖培养小鼠足细胞凋亡的影响

Effects of Zuogui Jiangtang Yishen Decoction Plasma on Apoptosis of Podocyte Cells Cultured in High Glucose Condition

目的探讨在高糖作用下,体外培养的小鼠足细胞凋亡及bax、bcl-2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,25 mmo L/L葡萄糖,10%空白血浆)、高糖组(B组,200 mmo L/L葡萄糖,10%空白血浆)、左归降糖益肾方含药血浆组(C组,200 mmo L/L葡萄糖,10%含药血浆),4-苯基丁酸(4-PBA)含药血浆组(D组,200 mmo L/L葡萄糖,10%含药血浆)。采用间接免疫荧光细胞化学法检测足细胞凋亡情况;MTT自动比色法检测足细胞活力,蛋白质印迹法和逆转录-聚合酶链反应法分别检测bax及bcl-2蛋白或mRNA表达水平的变化。结果与A组相比,B组足细胞活力降低,bax及bcl-2蛋白或mRNA的表达水平均升高(P<0.01);与B组相比,C组、D组足细胞活力升高,bax蛋白或mRNA的表达水平均降低(P<0.05或P<0.01),bcl-2蛋白或mRNA的表达水平均升高(P<0.01);与C组比,D组足细胞活力升高(P<0.05),但bax及bcl-2蛋白或mRNA的表达水平,差异均无统计学意义(P>0.05)。结论 bax及bcl-2蛋白或mRNA表达水平的升高,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影响bax及bcl-2蛋白或mRNA的表达水平,从而抑制足细胞凋亡。

Objective To investigate the expression levels of bax, bcl-2 and cell apoptosis in podocyte cells cultured in high glucose condition in vitro, and the regulation effects of Zuogui Jiangtang Yishen decoction plasma. Methods Cultured podocyte cells of mouses were divided into four groups： blank group（group A, 25 mmo L/L glucose, 10% blank plasma）, high glucose group（group B, 200 mmo L/L glucose, 10% blank plasma）, Zuogui Jiangtang Yishen decoction plasma group（Group C, 200 mmo L/L glucose, 10% containing medicine plasma）, and 4-Phenylbutyric acid（4-PBA） group（group D, 200 mmo L/L glucose, 10% containing medicine plasma）. The cell apoptosis were observed by immunofluorescence stainings. The cell viability was detected by MTT colorimetry assay. The protein and mRNA expressions of bax and bcl-2were analyzed by Western blot or reverse transcription-polymerase chain reaction, respectively. Results Compared with the group A, the viability of podocyte cells was significantly decreased of group B, and their expressions of bax and bcl-2protein or mRNA were upregulated（P〈0.01）. Compared with group B, the expressions of bax protein or mRNA of podocyte cells were downregulated（P 〉0.05 or P 〉0.01）, but the protein or mRNA of bcl-2 were upregulated（P 0.01）. Compared with group C, the cell viability of group D was significantly upcreased（P〈0.05）, but there was no statistical significance of their expressions of bax and bcl-2 protein or mRNA（P 〉0.05）. Conclusion The higher level expressions of bax and bcl-2protein or mRNA were the molecular mechanisms of podocyte injury. ZJYD plasma can repair the damaged podocyte by regulateing the protein or mRNA expressions of bax and bcl-2.