高糖对小鼠足细胞表型转化的诱导作用及机制探讨

摘要

目的 观察高糖对小鼠足细胞表型转化的诱导作用,并探讨其机制.方法 将传代培养后的小鼠足细胞随机分为A、B、C三组,A组常规培养,B组加入终浓度为30 mmol/L的葡萄糖、C组加入终浓度为30 mmol/L的葡萄糖及10 μmol/L的蛋白质酪氨酸激酶(JAK2)特异性阻断剂α-氰-(3,4-二羟基)-N-苄基肉桂酰胺(AG490)进行培养,48 h后收集细胞,采用Western blot法检测足细胞中自身标志物人肾病蛋白(Nephrin)、间充质细胞标志物α-平滑肌肌动蛋白(α-SMA)及磷酸化蛋白质酪氨酸激酶(p-JAK2)蛋白,采用RT-PCR检测足细胞中的Nephrin、α-SMA mRNA.结果 A组足细胞中Nephrin、α-SMA、p-JAK2蛋白积分光密度值分别为0.96 ±0.01、0.21 ±0.02、0.39±0.01,B组分别为0.58 ±0.01、0.66 ±0.07、0.71 ±0.02,C组分别为0.87±0.04、0.35 ±0.02、0.41±0.04;A组足细胞中Nephrin、α-SMA mRNA的相对表达量为1.53 ±0.04、0.57 ±0.01,B组分别为0.82 ±0.09、0.89±0.03,C组分别为1.30 ±0.04、0.78 ±0.03.B组与A组比较,P均＜0.05；C组与B组比较,P均＜0.05.结论 高糖可通过激活JAK/STAT信号途径诱导足细胞发生表型转化.%Objective To investigate the mechanism of high glucose-induced epithelial-mesenchymal transition in podocytes. Methods Cultured mouse podocytes were divided into three groups; group A ( cells cultured conventionally) , group B (cells cultured in 30 mmol/L glucose condition) and group C (3 cells cultured in 30 mmol/L glucose and 10 μmol/L AG490 conditions). Cells were collected after cultured 48 h. Western blotting analysis was used to determine the expression of nephrin, α-SMA and p-JAK2. RT-PCR analysis was used to detect the nephrin and α-SMA mRNA. Results The relative absorbance values of nephrin,α-SMA and p-JAK2 were 0.96 ±0.01 ,0.21 ±0.02,0.39 ±0.01 in group A, 0.58 ±0.01,0.66 ±0.07,0.71 ±0.02 in group B, 0.87 ±0.04,0.35 ±0.02,0.41 ±0.04 in group C. The relative absorbance values of nephrin,α-SMA mRNA were 1. 53 ± 0. 04,0. 57 ± 0. 01 in group A ,0. 82 ± 0. 09 ,0. 89 ± 0. 03 in group B,l. 30 ±0.04,0. 78 ±0.03 in group C. Compared with group B, these values in group A and C showed significant difference (all P<0. 05). Conclusion High glucose can induce the epithelial-mesenchymal transition in podocytes by activation of the JAK/STAT pathway.