局灶性节段性肾小球硬化患者血清对体外培养足细胞凋亡和黏附功能的影响

The effect of serum from FSGS patients on apoptosis and adherence of podocytes

目的探讨局灶性节段性肾小球硬化(FSGS)患者血清对体外培养足细胞凋亡和黏附功能的影响。方法提取原发性FSGS患者和正常人血清,分别以20%正常血清、5%、10%、20%FSGS血清孵育体外培养足细胞24h、48h,乳酸脱氢酶(LDH)释放实验检测足细胞活力,Hoechst33258法检测足细胞凋亡率,分光光度法检测足细胞caspase-3酶活性,并检测足细胞黏附能力。结果 FSGS患者血清呈时间和浓度依赖性地升高足细胞LDH释放率(与同时间点正常血清组比较,P分别<0.05和<0.01),凋亡率(与同时间点正常血清组比较,P分别<0.05和<0.01)及caspase-3活性(与同时间点正常血清组比较,P均<0.01),同时降低了足细胞的黏附功能(与同时间点正常血清组比较,P均<0.05)。结论 FSGS患者血清能诱导体外培养足细胞凋亡,且降低其黏附功能,导致足细胞的数目减少。

Objective To explore the effect of serum from FSGS patients on apoptosis and adherence of podocytes.Methods Serum from primary FSGS patients and normal volunteers were collected.Cultured podocytes were incubated with 20% normal serum from volunteers or 5%,10%,20% serum from FSGS patients for 24 h and 48 h,respectively.Podocyte viability,apoptosis,and enzymatic activity of caspase-3 were assessed by LDH release test,Hoechst 33258,spectrophometry respectively,adherent ability of podocytes was also detected.Results Serum from FSGS patients could increase LDH release（compared with normal sera group,P〈0.05 and0.01 for 24 h and 48 h time points respectively）,apoptosis ratio（compared with normal sera group,P〈0.05 and0.01 for 24 h and 48 h time points respectively）and caspase-3 activity（compared with normal sera group,P〈0.01 for both time points）,but decrease podocyte adherent ability in time and concentration-dependent manners（compared with normal sera group,P〈0.05 for both time points）.Conclusions Serum from FSGS patients may induce apoptosis and reduce adherence of podocye in vitro,which likely contributes to the podocyte loss in patients with FSGS.