吡格列酮对自发性高血压大鼠心脏成纤维细胞周期的影响

Effects of Pioglitazone on cell cycle of cardiac fibroblasts in spontaneous hypertesion rats

目的研究盐酸吡格列酮对培养的自发性高血压大鼠(SHR)和血压正常大鼠(WKY)心脏成纤维细胞(CFs)细胞周期的影响,并进一步探讨其与NO-NOS系统的关系。方法采用胶原酶消化法培养SHR和WKY的CFs,流式细胞仪(FCM)分析技术检测细胞周期,硝酸还原酶法测定CFs培养上清NO浓度,分光光度法测定CFs培养上清NOS活性。结果0.1、1、5和10μmol/L的吡格列酮作用48h后,SHR、WKY CFs的G0/G1期细胞百分率较对照组显著增高(均P<0.01),S期细胞百分率、G2/M期细胞百分率和增殖指数则显著低于对照组(P<0.01,P<0.05),且随着作用浓度的增加,吡格列酮对细胞周期的影响逐渐增强。5μmol/L的吡格列酮干预48 h后,SHR和WKY的CFs培养上清NO浓度分别为(111.2±12.4)μmol/L和(221.7±35.3)μmol/L,与各自对照组(76.8±2.4)μmol/L、(112.1±8.9)μmol/L相比,差异非常显著(P<0.01);SHR和WKY大鼠CFs培养上清NOS活性分别为(208±18)μkat/L、(257±30)μkat/L,和各自对照组(148±10)μkat/L、(187±8)μkat/L相比,亦有非常显著性差异(P<0.01)。NO浓度和NOS活性呈显著正相关(r=0.964,P<0.01)。结论吡格列酮能够抑制SHR CFs的细胞周期增殖,其效应可能与上调NO-NOS系统活性有关。

AIM To evaluate the effects of insulin-sensitizer Pioglitazone on the cell cycle of cardiac fibroblasts （CFs） isolated from SHR and WKY rat and the activity of NO-NOS. METHODS Isolated and cultured CFs of SHR and WKY rats were used as experiment model. Cell cycle was determined by FCM. Nitrate enzyme reverting methods and spectrophotometer were also adopted to detect the content of NO and activity of NOS in CFs of SHR and WKY rats with or without Pioglitazone. RESULTS Based on the analysis of FCM cell cycle, it was shown that the percentage of cells on S and PI stages in CFs of SHR and Wistar was gradually declined as Pioglitazone concentration increased, while the percentage of cells on G0/G1 stages rose（P 〈 0.01 ）. In SHR and Wistar rats, the NO concentration in supematant （111.2± 12.4） μmol/L, （221.7± 35.3 ） μmol/L, respectively treated by 5 μmol/L pioglitazone for 48 hours, was significantly higher than that of control group （76.8 ± 2.4） μmol/L, （ 112. 1± 8.9）μmol/L, respectively, P 〈 0. 01. NOS activity in supernatant of SHR and Wistar rats [ （208 ±18 ） μkat/L, （257±30） μkat/L, respectively ] treated by 5 μmol/L pioglitazone for 48 hours, was also significantly higher than that of control group （148 ±10） μkat/L, （187 ±8）μkat/L, respectively, P 〈0. 01. There was a significantly positive correlation between NO contents and NOS activity （r = 0. 964, P 〈 0. 01 ）. CONCLUSION Insulin-sensitizer hydrochloride Pioglitazone inhabits the cell cycle proliferation of CFs of SHR in vitro, which may related with the up-regulation of NO-NOS system.