探讨茶黄素对糖尿病肾病大鼠肾小球系膜细胞p38丝裂原活化蛋白激酶（p38MAPK）及细胞外基质(ECM)合成的影响。分别以正常糖（NG）、高糖（HG）、糖基化终产物（AGE）及过氧化氢（H2O2）孵育大鼠肾小球系膜细胞,Western Blot检测系膜细胞中p38MAPK和转化生长因子-β(TGF-β)蛋白表达,酶联免疫吸附实验（ELISA）检测细胞培养液中细胞外基质成分纤维连接蛋白（FN）、Ⅳ型胶原（Col Ⅳ）及层黏连蛋白（LN）的含量。结果显示HG、AGE和H2O2能明显诱导p38MAPK磷酸化和TGF-β蛋白表达增加而导致细胞外基质的积聚,茶黄素能明显抑制p38MAPK的磷酸化活性和TGF-β蛋白表达及减少细胞外基质的积聚。提示茶黄素可通过调节p38MAPK信号转导通路而减少细胞外基质的合成,延缓糖尿病肾小球肥大和肾小球硬化。
To study the effect of theaflavin on rat mesangial cells (GMCs) were separately incubated with normal glucose, high glucose, advanced glycosylation end products (AGE) and H2O2. The protein expression of p-p38MAPK and TGF-β was examined by Western blot, and the levels of fibronectin (FN), collagen Ⅳ(ColⅣ), laminine (LN) in the supernatant of cultured GMCs were detected by ELISA. Compared to the control group, high glucose, AGE and H2O2 significantly activated p-p38MAPK and increased the protein expression of TGF-β and extracellar matrix production. The protein expression of p-p38MAPK and TGF-β was inhibited by theaflavin. Compared to GMCs incubated with corresponding stimulators, FN, ColⅣ and LN were expressed at a lower level after pre-treatment of theaflavin. It can be concluded that theaflavin may effectively attenuate diabetic nephropathy progression through down-regulating the expression of TGF-β and extracellar matrix production via p38MAPK.
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