【摘要】 目的观察黄芪甲苷(AS-Ⅳ)对高糖环境下足细胞黏附能力的影响,并探讨其作用机制。方法将条件永生化小鼠足细胞分为正常对照组、高糖组、高糖+低剂量AS-Ⅳ组、高糖+高剂量AS-Ⅳ组,采用细胞黏附荧光定量试剂盒检测足细胞黏附能力,白蛋白流量率检测法测定足细胞单层屏障功能,Western blot法检测粘着斑激酶(FAK)与磷酸化FAK(p-FAK)表达。观察并比较4组足细胞黏附能力、单层屏障功能、FAK与p-FAK表达情况。结果 4组足细胞黏附能力、单层屏障功能比较差异均有统计学意义(均P<0.05),高糖环境会降低足细胞的黏附能力、单层屏障功能,而AS-Ⅳ可明显增强高糖环境下足细胞的黏附能力、单层屏障功能,且对黏附能力的影响具有时间和剂量依赖性。4组足细胞FAK表达情况比较无统计学差异(均P>0.05),而p-FAK表达情况比较有统计学差异(均P<0.05);高糖环境对足细胞FAK表达无明显影响,但会明显增加FAK磷酸化;AS-Ⅳ可明显抑制高糖环境下足细胞FAK蛋白磷酸化,且该作用具有时间和剂量依赖性。结论 AS-Ⅳ或通过抑制FAK磷酸化改善高糖环境下足细胞的黏附能力。更多还原

【Abstract】 Objective To investigate the effect of astragaloside Ⅳ(AS-Ⅳ) on podocyte adhesion function under high glucose circumstances and explore its mechanisms. Methods Conditionally immortalized mouse podocytes were divided into normal control group,high glucose group, high glucose with low dose AS-Ⅳ group and high glucose with high dose AS-Ⅳ group. Podocyte adhesion was measured by fluorescence. The filtration barrier function of podocyte monolayer was measured by albumin influx. The expression of focal adhesion kinase(FAK) and phosphorylated FAK(p-FAK) were measured by Western blot. The results of 4 groups were compared. Results The adhesion ability and monolayer barrier function of podocytes among 4 groups were significantly different(all P<0.05). Compared with normal control group podocyte adhesion with basement membrane complex and monolayer filtration barrier function reduced significantly under high glucose circumstances and were improved by AS-Ⅳ. AS-Ⅳ attenuated podocyte adhesion reduction induced by high glucose in a time and dose dependent manner. The protein expression of FAK had no significant difference among 4 groups(all P >0.05), but there were significant differences in p-FAK expression among 4 groups(all P <0.05). The p-FAK expression in podocytes was increased significantly under high glucose circumstances, and was inhibited by AS-Ⅳ time-and dose-dependently. Conclusion The study indicates that AS-Ⅳ may improve podocyte adhesion function in diabetic nephropathy, which may be associated with the inhibition of FAK phosphorylation.更多还原