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目的 在周期素激酶抑制剂p2 7和血管紧张素Ⅱ受体 1(AT1)水平 ,研究血管紧张素Ⅱ(AngⅡ )诱导系膜细胞 (MC)肥大的分子机制。方法 用蛋白印迹 (western杂交 )方法测定MC中p2 7蛋白水平 ;用 [3 H]胸腺嘧啶核苷 ([3 H]TdR)及 [3 H]亮氨酸 ([3 H]leu)掺入方法测定MC的肥大情况 ,观察p2 7反义寡核苷酸 (ODN)转染对MC肥大程度的影响 ;用ELISA方法测定MC中细胞外基质 (ECM)蛋白 (Ⅳ型胶原及纤维连接蛋白 )。结果 AngⅡ使无血清培养MC中p2 7增多 ,[3 H]leu掺入增加 ,[3 H]TdR掺入减少 ,MC中ECM水平增高 ,p2 7反义ODN转染使AngⅡ的上述作用减弱 ;氯沙坦可降低AngⅡ刺激的MC中的p2 7水平 ,减少 [3 H]leu掺入 ,增加 [3 H]TdR掺入 ,降低MC中的ECM水平 ,且上述作用呈剂量依赖性。结论 AngⅡ通过AT1受体可提高p2 7水平 ,诱导MC细胞肥大 ,而氯沙坦可减轻AngⅡ诱导的MC细胞肥大程度。
Objective To investigate the molecular mechanism of angiotensin Ⅱ induced mesangial cell (MC) hypertrophy at the p2 7 and AT1 levels. Methods The level of p27 protein in MC was determined by Western blot (Western blot). The level of p27 protein in MC was measured by using [3 H] thymidine ([3 H] TdR) and [3 H] leucine Methods The hypertrophy of MC was observed to observe the effect of p27 antisense oligodeoxynucleotide (ODN) transfection on MC hypertrophy. The expression of extracellular matrix (ECM) protein (type Ⅳ collagen and fibronectin) . Results Ang Ⅱ increased the expression of p27 in MC without serum, increased the incorporation of [3 H] leu, decreased the incorporation of [3 H] TdR and increased the level of ECM in MC. The above effect of AngⅡ was attenuated by p2 7 antisense ODN transfection. Losartan decreased the level of p27 in Ang II-stimulated MCs, decreased [3 H] leu incorporation, increased [3 H] TdR incorporation, and decreased ECM levels in MCs in a dose-dependent manner. Conclusion AngⅡ can increase p27 level through AT1 receptor and induce MC cell hypertrophy. Losartan can reduce Ang Ⅱ-induced MC cell hypertrophy.