【作者】 杜婧；

【导师】 叶传蕙；

【作者基本信息】 成都中医药大学 ， 中西医结合临床内科， 2003， 博士

【摘要】 目的：观察中药复方制剂肾康注射液对高糖培养大鼠肾小球系膜细胞周期异常的调节作用及其机制，探讨肾康注射液对早期DN的防治作用。 方法：将体外培养的正常大鼠肾小球系膜细胞分为7组，分别为正常组、高糖组、甘露醇组、苯那普利组和不同剂量肾康注射液治疗组，每组设3个复孔，共培养72小时。应用流式细胞术观察细胞周期分布、RNA/DNA比值、细胞大小；ELISA法测定培养细胞上清液中FN的含量；RT-PCR检测各组细胞TGF-β₁mRNA、P²¹mRNA的表达。 结果：（1）高糖培养肾小球系膜细胞出现细胞周期分布异常，G₁期细胞比例增高，G₂期、S期比例下降，RNA/DNA比值增高，以及细胞体积增大；肾康注射液高剂量组可以使细胞周期分布、RNA/DNA比值趋于正常，并能抑制细胞体积增大。（2）高糖培养系膜细胞FN合成及分泌增多，肾康注射液高剂量组能抑制FN的过度合成及分泌。（3）高糖培养系膜细胞TGF-β₁mRNA、P²¹mRNA表达增加，肾康注射液可以抑制TGF-β ImRNA、P²¹mRNA过度表达，高剂量组作用优于苯那普利。 结论：（1）高糖培养肾小球系膜细胞出现细胞周期异常改变，表现为细胞停滞在G₁期，细胞肥大，FN合成及分泌增多；肾康注射液可以改善细胞周期异常改变，促使细胞周期正常运转，肥大减轻，FN异常合成及分泌减少。（2）肾康注射液在转录水平影响系膜细胞TGF-β₁以及P²¹合成，从而促使细胞周期正常运转，可能是肾康注射液调节高糖培养肾小球系膜细胞周期异常的重要机理。更多还原

【Abstract】 Objective: To investigate effects and mechanisms of Shenkang Injection, a compound TCM preparation, on abnormal cell cycle of glomerular mesangial cells cultured in high glucose and explore the effects of Shenkang Injection to prevent and treat diabetic nephropathy.Methods: Cultured rat glomerular mesangial cells were diviced into 7 groups: comtrol group, High Glucose-treated group, Mannitol-treated group, Benazepril-treated group, Shenkang Injection-treated groups with different dose. Every group was trested for 72 hours. The cell cycle , RNA/DNA and cells vovulme of glomerular mesangial cells were measured by flow cytometry. The level of fibronectin in the cultured supernatant was measured by ELISA . The expression of TGF- 1 mRNA and P21 mRNA were examined by RT-PCR.Results: (1) High glucose caused the abnormal cell cycle of glomerular mesangial cells as follows: cells proportion in Gl phase increased, cell proportion in G2 and S phase detreased, RNA/DNA up-regulated, cellular size enlarged. The high-dose Shenkang Injection could decrease cells proportion in phase Gl, increase that in phase G2 and S, and reduce RNA/DNA and cellular size. (2) High glucose increase the level of FN in glumerular measangial cella. The High-dose Shenkang Injection could ruduce the levelof FN. (3)High glucose increased the expression of TGF-31mRNA and P21mRNA , Shenkang Injection could inhibit the overexpression of TGF- PlmRNA and P21mRNA induced by high glucose. High-dose The effect of Shenkang Injection is superio to Benazepril.Conclusion: (1) The abnormal cell cycle of high-glucose cultured glomerular mesangial cells presented retention of cell division at Gl phase, RNA/DNA increase, cells hypertrophy, FN synthesis and secretion increase; Shenkang Injection was able to improve abnormal cell cycle and regulate cell proportion of different phases. In addition, it could reduce hypertrophy of cells, FN synthesis and secretion. (2) TGF- PlmRNA and P21mRNA synthesis of mesangial cells were regulated at transcription level by Shengkang Injection. Consequently, normal cell cycle, were obtained. This may be the important regulative mechanism of Shenkang Injection on abnormal cell cycle of glomerular mesangial cells in high glucose.更多还原