缬沙坦抑制高糖环境下足细胞损伤的机制探究

Study on the mechanism of valsartan inhibiting podocytes injury in high glucose environment

目的通过观察缬沙坦对高糖环境下足细胞EMT过程中ILK、MMP9、NEPH1 mRNA表达的影响,探讨缬沙坦保护受高糖损伤足细胞的可能机制。方法将培养分化成熟的足细胞随机分为5 mmol/L葡萄糖培养组(对照组)和25 mmol/L葡萄糖培养组(高糖组),在25 mmol/L葡萄糖培养液中分别加入2×10^(-7)mol/L(低Val组)、2×10^(-6)mol/L(中Val组)和2×10^(-5) mol/L(高Val组)缬沙坦。体外培养48 h后,倒置显微镜观察细胞形态,并采用PCR半定量分析技术检测ILK、MMP9、NEPH1的表达。结果高糖组细胞NEPH1的表达较对照组显著减少(P<0.01)。而各剂量缬沙坦干预组NEPH1的表达与高糖组相比均显著升高。与对照组比较,高糖组足细胞ILK和MMP9的表达显著升高,而各剂量缬沙坦干预组足细胞ILK和MMP9的表达对比高糖组均明显减少(P<0.01),其中以高Val组的干预作用最显著(P<0.01),高Val组ILK和MMP9的表达与对照组无明显差异(P>0.05)。结论缬沙坦可能通过抑制ILK通路保护高糖环境下受损的足细胞。

Objective To observe the effect of valsartan on ILK, MMP9 and NEPH1 mRNA expression during the EMT process of podocytes under high glucose environment, and to explore the possible mechanism of valsartan in protecting the podocytes from high glucose. Methods The differentiated mature podocytes were randomly divided into 5 mmol/L glucose culture group （control group） and 25 mmol/L glucose culture group （high glucose group）. 2×10^-7mol/L valsartan （low Val group）, 2×10^-6 mol/L valsartan （medium Val group） and 2×10^-5 mol/L （high Val group） valsartan were added to 25 mmol/L glucose culture medium. After 48 hours of in vitro culture, the cell morphology was observed by inverted microscope, and the expression of ILK, MMP9 and NEPH1 were detected by PCR semi-quantitative analysis. Results The expression of NEPH1 in high glucose group was significantly lower than that in control group（P〈0.01）. The expression of NEPH1 in each dose of valsartan intervention group was significantly higher than that in high glucose group. Compared with the control group, the expression of ILK and MMP9 of podocytes in the high glucose group was significantly increased, and the expression of ILK and MMP9 of podocytes in each dose of valsartan intervention group was significantly lower than that in high glucose group （P〈0.01）. The intervention effect in high Val group was the most significant（P〈0.01）. The expression of ILK and MMP9 in high Val group was not significantly different from that in control group（P〉0.05）. Conclusion Valsartan may protect the podocytes from the high glucose environment by inhibiting the ILK pathway.