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10μmol/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 10μmol/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 groupB, 0. 87 ± 0. 04 ,0. 35±0. 02 ,0. 41±0. 04 in group C. The relativeab-sorbance 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, 1.30±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.
出处
《山东医药》
CAS
2012年第34期22-24,共3页
Shandong Medical Journal
基金
河北省教育厅科学研究项目(2011168)
河北省卫生厅医学科学研究重点课题资项目(20110081)
