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本文旨在观察p38MAPK与高糖诱导的肾小管上皮细胞向间充质细胞转变之间的关系。将雄性Sprague-Dawley(SD)大鼠随机分为对照组、糖尿病组、胰岛素治疗组,用免疫组织化学、Westernblot检测p38MAPK和磷酸化p38MAPK(p-p38MAPK)蛋白表达。采用机械分离和酶消化获取SD大鼠肾小管节段,进行肾小管上皮细胞培养,将肾小管上皮细胞分为对照组、高渗组(20mmol/LD-mannitol)、高糖组(20mmol/LD-glucose)和SB202190(p38MAPK特异性抑制剂)+高糖组,处理72h后收集细胞,用免疫细胞化学检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、p-p38MAPK和Snail1蛋白表达,Western blot检测p38MAPK、p-p38MAPK、Snail1、转化生长因子β1(transforming growth factor-β1,TGF-β1)、α-SMA和E-cadherin的表达,RT-PCR检测α-SMA和E-cadherin mRNA的表达。体内和体外结果均显示,高糖状态激活了p38MAPK,这种活化作用在体内可因胰岛素控制血糖而被消除,在体外可被p38MAPK特异性抑制剂SB202190显著抑制;高糖组α-SMA蛋白和mRNA在原代培养肾小管上皮细胞的表达较对照组分别增加12倍和8倍(P<0.01),SB202190处理组其表达则较高糖组分别减少67%和50%(P<0.01)。SB202190不影响TGF-β1蛋白表达,但下调Snail1蛋白表达,并部分恢复高糖组E-cadherin蛋白和mRNA的表达。上述结果提示,p38MAPK可能通过转录因子Snail1介导高糖诱导的肾小管上皮细胞向间充质细胞转变。
This article aims to observe the relationship between p38 MAPK and high glucose-induced renal tubular epithelial cells to mesenchymal transition. Male Sprague-Dawley (SD) rats were randomly divided into control group, diabetes group and insulin treatment group. The protein expressions of p38MAPK and p38MAPK (p-p38MAPK) were detected by immunohistochemistry and Western blot. The renal tubular epithelial cells were harvested by mechanical separation and enzymatic digestion. The renal tubular epithelial cells were divided into control group, hypertonic group (20 mmol / L-mannitol), high glucose group (20 mmol / LD -Glucose and SB202190 (specific inhibitors of p38MAPK) + high glucose group. The cells were harvested after 72 hours and the expressions of α-smooth muscle actin (α-SMA), p-p38MAPK and The expressions of Snail1 and p-p38MAPK, Snail1, TGF-β1, α-SMA and E-cadherin were detected by Western blot. The expressions of α-SMA and E-cadherin -cadherin mRNA expression. Both in vivo and in vitro results show that p38MAPK is activated by high glucose, which can be abolished by the control of blood glucose in the body and inhibited by p20MAPK specific inhibitor SB202190 in vitro. The α-SMA protein and Compared with the control group, the expression of mRNA in primary cultured renal tubular epithelial cells increased by 12-fold and 8-fold (P <0.01), while the expression of SB202190 decreased by 67% and 50% (P <0.01), respectively. SB202190 did not affect the expression of TGF-β1, but down-regulated the expression of Snail1 protein and partially restored the expression of E-cadherin protein and mRNA in high glucose group. These results suggest that p38MAPK may mediate high glucose-induced tubular epithelial-to-mesenchymal transition through the transcription factor Snail1.