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目的探讨高糖培养环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和基质金属蛋白酶9(MMP-9)表达的影响。方法体外培养的小鼠肾小球足细胞分为空白对照组、高渗对照组、高糖组和高糖+雷帕霉素干预组。Real-time PCR和Western blotting检测各组肾小球足细胞Ⅳ型胶原α3链(COL4A3)、α5链(COL4A5)mRNA及MMP-9mRNA和蛋白的表达;ELISA法检测细胞培养上清液中Ⅳ型胶原的含量。结果与对照组比较,高糖组足细胞COL4A3、COL4A5mRNA表达及培养上清液中Ⅳ型胶原含量均升高(P<0.05),而足细胞MMP-9mRNA和蛋白表达均降低(P<0.05)。与高糖组比较,雷帕霉素干预组足细胞COL4A3、COL4A5mRNA表达及培养上清液中Ⅳ型胶原含量均降低(P<0.05),足细胞MMP-9mRNA和蛋白表达均升高(P<0.05)。结论雷帕霉素对高糖环境中足细胞Ⅳ型胶原和MMP-9表达异常具有逆转作用。
Objective To investigate the effects of high glucose culture environment and rapamycin on the expressions of type Ⅳ collagen and matrix metalloproteinase-9 (MMP-9) in glomerular podocytes. Methods The mouse glomerular podocyte cultured in vitro was divided into blank control group, hypertonic control group, high glucose group and high glucose + rapamycin intervention group. Real-time PCR and Western blotting were used to detect the expression of COL4A3 and COL4A5 mRNA and MMP-9 mRNA and protein in glomerular podocytes. The expression of type Ⅳ Collagen content. Results Compared with the control group, the expression of COL4A3, COL4A5 mRNA and the content of type Ⅳ collagen in the supernatant of the high glucose group were significantly increased (P <0.05), while the expressions of MMP-9 mRNA and protein in the podocytes were decreased (P <0.05) . Compared with high glucose group, the expressions of COL4A3 and COL4A5mRNA in podocytes and the content of type Ⅳ collagen in supernatant of rapamycin group were significantly decreased (P <0.05), while the mRNA and protein expressions of MMP-9 in podocytes were increased (P < 0.05). Conclusion Rapamycin can reverse the abnormal expression of type Ⅳ collagen and MMP-9 in podocytes in high glucose environment.