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目的 探讨黄葵浸膏粉对高糖诱导下足细胞相关蛋白Nephrin和Podocin表达的影响及其在糖尿病肾病治疗中的作用机制.方法 以小鼠足细胞系(MPC5)为研究对象,以正常糖浓度(NG,5.6 mmol/L)为对照,将培养好的足细胞置于高糖浓度(HG,25 mmol/L)和不同处理浓度的黄葵浸膏粉(HK,0 g/L、0.45 g/L)中处理24 h,将其分为四组:5.6 mmol/L NG组、5.6 mmol/L NG+ 0.45 g/L HK组、25 mmol/L HG组、25 mmol/L HG+ 0.45g/L HK,其中以5.6 mmol/L NG为对照组.采用细胞凋亡双染试剂盒(Annexin-v FITC/PI)双染法检测细胞凋亡、定量即时聚合酶链锁反应(qRT-PCR)及western blot检测Nephrin和Podocin的mRNA和蛋白表达水平.结果 与对照组比较,25 mmol/L HG组的足细胞凋亡率[(20.39±0.03)％比(17.70±0.91)％]显著增高(t=2.947,P＜0.05);在高糖浓度处理下,与25 mmol/L HG组比较,25 mmol/L HG+ 0.45 g/L HK组的足细胞凋亡率[(11.96±1.11)％比(20.39±0.03)％]显著降低(t=7.586,P＜0.01).qRT-PCR及western blot检测结果发现,与对照组比较,高糖浓度显著抑制了Nephrin和Podocin的表达[(0.489±0.040)比(0.721 ±0.022),t =4.992,P＜ 0.01;(0.387±0.014)比(0.778±0.036),t=10.050,P＜0.01];在高糖浓度处理条件下,适当浓度的黄葵浸膏粉可促进足细胞相关蛋白Nephrin和Podocin的表达[(0.603 ±0.013)比(0.489±0.040),t=2.653,P ＜0.05;(0.640±0.024)比(0.387±0.014),t=8.946,P＜0.01].结论 长时间的高糖浓度处理可诱导足细胞凋亡,但一定浓度的黄葵浸膏粉可抑制高糖诱导的足细胞凋亡.在高糖浓度下,黄葵浸膏粉可能通过调控足细胞相关蛋白Nephrin和Podocin的表达,从而抑制足细胞的凋亡,对足细胞起着重要的保护作用.“,”Objective To investigate the effect of Huangkui extract powder (HK) on the expression of nephrin and podocin proteins in mouse podocytes induced by high glucose,which is involved in the treatment of diabetic nephropathy (DN).Methods Cultured mouse podocytes (MPC5) were incubated in high glucose and HK at 5.6 mmol/L NG,5.6 mmol/L NG + 0.45 g/L HK,25 mmol/L HG,25 mmol/L HG + 0.45 g/L HK,respectively.The 5.6 mmol/L NG group was used as normal control.After 24 hours of intervention,we detected podocyte apoptosis by Annexin-V FITC/PI double staining,measured the mRNA and protein expression of nephrin and podocin by qRT-PCR and Western blot.Results Compared with the control group (5.6 mmol/L,NG),the apoptosis rate of podocytes in the high glucose concentration group (25 mmol/L,HG) was significantly higher [(20.39 ± 0.03) ％ vs.(17.70 ± 0.91) ％,t =2.947,P ＜ 0.05)].The apoptosis rate of podocytes in the 25 mmol/L HG + 0.45 g/L HK group was significantly lower than that in the 25 mmol/L HG group [(11.96 ± 1.11) ％ vs.(20.39 ± 0.03) ％,t =7.586,P ＜ 0.01].The results of qRT-PCR and Western blot showed that the expression of nephrin and podocin was significantly inhibited by high glucose concentration compared with the control group[(0.489 ±0.040) vs.(0.721 ±0.022),t =4.992,P ＜0.01;(0.387 ±0.014) vs.(0.778 ±0.036),t =10.050,P ＜0.01],and the expression of podocin and nephrin was increased by appropriate concentration of H K [(0.603 ± 0.013) vs.(0.489 ± 0.040),t =2.653,P＜0.05;(0.640±0.024) vs.(0.387 ±0.014),t=8.946,P＜0.01].Conclusion Podocyte apoptosis can be induced by prolonged high glucose treatment,but a certain concentration of HK can inhibit podocyte death induced by high glucose.The possible mechanism is that HK may inhibit the apoptosis of podocytes by regulating the expression of podocin and nephrin in podocytes at high glucose concentration,thus plays a protective role on podocytes.