Effect of Huangkui extract powder on the expression of nephrin and podocin in podocyte induced by high glucose / 中国基层医药

ABSTRACT

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.