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目的观察脂多糖( LPS)对足细胞活动力的作用,探讨环孢素(CsA)对脂多糖诱导足细胞活动力改变的影响。方法足细胞分为3组:正常对照组, LPS组(50mg·L-1 LPS作用足细胞24h), LPS+CsA组(50mg·L-1 LPS与0.5mg·L-1CsA共同作用足细胞24h)。利用免疫荧光、流式细胞术、转板迁移测定法等,检测整合素β3分布和活化情况以及足细胞活动力的变化。结果3组足细胞整合素β3分布未见显著差异。抗活化整合素β3的AP5抗体表达率、转板迁移测定法中每个视野足细胞数 LPS组[(52.47±s0.12)%,(328±24)个]均高于正常对照组[(1.25±0.12)%,(177±7)个]和 LPS+CsA组[(10.28±0.28)%,(232±10)个](P<0.01), LPS+CsA组均高于正常对照组(P<0.01)。与正常对照组相比,环孢素干预后整合素β3的活化水平仍然较高(P<0.01),但2组迁移能力并无显著差异(P>0.05)。结论脂多糖诱导足细胞损伤时,整合素β3的活化、足细胞的活动增强。环孢素可能通过降低整合素β3活化,降低足细胞活动力,稳定足细胞。
Objective To observe the effect of lipopolysaccharide (LPS) on podocyte viability and to explore the effect of cyclosporine (CsA) on the changes of podocyte viability induced by lipopolysaccharide. Methods Podocytes were divided into 3 groups: normal control group, LPS group (50mg · L-1 LPS-treated podocytes for 24h), LPS + CsA group (50mg · L-1 LPS and 0.5mg · L-1CsA) ). Immunofluorescence, flow cytometry, plate transfer assay and so on, to detect integrin β3 distribution and activation and changes in podocyte viability. Results There was no significant difference in integrin β3 distribution between the three groups. The AP5 antibody expression rate of anti-activated integrin β3 was higher in LPS group [(52.47 ± s0.12)%, (328 ± 24)%] than that in normal control group [ (P <0.01), LPS + CsA group (1.25 ± 0.12)%, (177 ± 7) and LPS + CsA group (10.28 ± 0.28)% and (232 ± 10) P <0.01). Compared with the normal control group, the activation level of integrin β3 was still high after cyclosporine intervention (P <0.01), but there was no significant difference between the two groups in migration ability (P> 0.05). Conclusion LPS can induce the activation of integrin β3 and increase the activity of podocytes when podocyte injury is induced. Ciclosporin may stabilize podocytes by decreasing the activation of integrin β3 and decreasing the podocyte viability.