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目的观察PI3K/Akt在高糖诱导的肾小管上皮细胞转分化中的作用及对分化抑制因子2(Id2)表达的影响。方法大鼠肾小管上皮细胞随机分成3组:对照(Con)组,高糖(H-Glu)组和PI3K抑制剂Wortmannin(Wort)组。Con组加正常培养液,H-Glu组在培养液中加30 mmol/L葡萄糖,Wort组用Wortmannin预处理1 h后,加30 mmol/L葡萄糖。24 h后用免疫细胞化学、Western blot法和RT-PCR法检测α平滑肌肌动蛋白(α-SMA)、p-Akt和Id2的表达。结果 Con组α-SMA、p-Akt阳性表达细胞很少,绝大部分细胞出现Id2阳性表达。与Con组比较,H-Glu组出现α-SMA和p-Akt表达升高,Id2表达降低(P<0.05)。与H-Glu组比较,Wort组α-SMA和p-Akt表达下降,Id2表达升高(P<0.05)。结论 PI3K/Akt可能通过抑制Id2基因表达,参与高糖诱导的大鼠肾小管上皮细胞转分化。
Objective To observe the effect of PI3K / Akt on high glucose induced transdifferentiation of renal tubular epithelial cells and its effect on the expression of differentiation inhibitor 2 (Id2). Methods Rat renal tubular epithelial cells were randomly divided into 3 groups: control group, H-Glu group and PI3K inhibitor Wortmannin group. Con group added normal culture solution, H-Glu group added 30 mmol / L glucose in culture medium, Wortmannin pretreatment for 1 h, added 30 mmol / L glucose. After 24 h, the expressions of α-SMA, p-Akt and Id2 were detected by immunocytochemistry, Western blot and RT-PCR. Results Con α-SMA, p-Akt positive cells rarely, most of the cells Id2 positive expression. Compared with Con group, the expression of α-SMA and p-Akt increased and the expression of Id2 decreased in H-Glu group (P <0.05). Compared with H-Glu group, the expression of α-SMA and p-Akt decreased and the expression of Id2 increased in Wort group (P <0.05). Conclusion PI3K / Akt may be involved in the high glucose-induced rat renal tubular epithelial cell transdifferentiation by inhibiting Id2 gene expression.