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【目的】研究盐酸吡格列酮对Wistar大鼠心脏成纤维细胞(CFs)细胞周期的影响及其与一氧化氮-一氧化氮合酶(NO-NOS)系统的关系。【方法】采用胶原酶消化法培养Wistar大鼠的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48 h后,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(均P<0.01),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,CFs培养上清NO浓度为221.7±35.3μmol/L,与对照组(112.1±8.9μmol/L)相比,差异非常显著(P<0.01);NOS活性为256.7±30.1 nkat/mL,和对照组(186.7±8.3 nkat/mL)相比,差异亦有非常显著性(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CFs的细胞周期增殖,其效应可能与上调NO-NOS系统活性有关。
【Objective】 To investigate the effect of pioglitazone hydrochloride on cell cycle of cardiac fibroblasts (CFs) in Wistar rats and its relationship with nitric oxide-nitric oxide synthase (NO-NOS) system. 【Methods】 The CFs of Wistar rats were cultured by collagenase digestion and the cell cycle was detected by flow cytometry (FCM). Nitric acid reductase was used to determine the concentration of NO in culture supernatant of CFs. The activity of NOS in supernatant of CFs was determined by spectrophotometry . 【Results】 After treated with 0.1, 1, 5 and 10 μmol / L pioglitazone for 48 h, the percentages of CFs in G0 / G1 phase were significantly higher than those in control group (all P <0.01). The percentages of cells in S phase and G2 / M phase And proliferation index were significantly lower than those in the control group (all P <0.01). With the increase of the concentration, the effect of pioglitazone on cell cycle increased gradually. Compared with the control group (112.1 ± 8.9μmol / L), the concentration of NO in the culture supernatant of CFs was 221.7 ± 35.3μmol / L after 48h treatment with 5μmol / L pioglitazone significantly (P <0.01). The activity of NOS was 256.7 ± 30.1 nkat / mL, compared with the control group (186.7 ± 8.3 nkat / mL), the difference was also significant (P <0.01). There was a significant positive correlation between NO concentration and NOS activity (r = 0.964, P <0.01). 【Conclusion】 Pioglitazone can inhibit cell cycle proliferation of CFs, which may be related to the up-regulation of NO-NOS activity.