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目的:研究广枣总黄酮(total?flavones?of?fructus?chorspondiatis,TFFC)对血管紧张素Ⅱ(angiontensin,AngⅡ)诱导大鼠心脏成纤维细胞(cardiac?fibroblasts,CFs)细胞周期的影响,并进一步探讨其与一氧化氮(nitric?oxide,NO)-一氧化氮合酶(nitricoxide?synthase,NOS)系统的关系。方法:建立新生大鼠心脏成纤维细胞系;采用流式细胞仪(flow?cytometry,FCM)分析技术检测细胞周期;采用化学比色法测定CFs培养上清乳酸脱氢酶(lactate?dehydrogenase,LDH)水平;采用硝酸还原酶法测细胞培养液中NO水平;化学比色法测细胞上清液中NOS水平。结果:25、50、100?mg/L的TFFC作用72?h后,CFs的G0/G1期细胞百分率较AngⅡ组显著增高(P<0.01,P<0.05),S期细胞百分率,G2/M期细胞百分率和增殖指数(proliferation?index,PI)则显著低于对照组(P<0.01,P<0.05),且随着作用浓度的增加,TFFC对细胞周期的影响逐渐增强。100?mg/L的TFFC干预72?h后,CFs培养上清NO浓度为(23.23±0.70)μmol/L,与AngⅡ组(20.20±0.83)μmol/L相比,差异非常显著(P<0.01);培养上清NOS活性为(20.43±0.53)U/L,和AngⅡ组(20.90±0.85)U/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:TFFC能够抑制AngⅡ诱导大鼠?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 total flavones of fructus chorspondiatis (TFFC) on the cell cycle of rat cardiac fibroblasts induced by angiotensin Ⅱ Further explore its relationship with nitric oxide (NO) - nitric oxide synthase (NOS) system. Methods: Newborn rat cardiac fibroblast cell line was established. Flow cytometry (FCM) was used to detect cell cycle. Chemiluminescence method was used to determine the expression of lactate dehydrogenase (LDH) ), The level of NO in cell culture medium was measured by nitrate reductase method, and the level of NOS in cell supernatant was measured by chemical colorimetry. Results: After treated with 25, 50, 100 mg / L TFFC for 72 h, the percentages of CFs in G0 / G1 phase were significantly higher than those in AngⅡ group (P <0.01, P <0.05) The percentage of staging cells and proliferation index (PI) were significantly lower than that of the control group (P <0.01, P <0.05), and the effects of TFFC on the cell cycle gradually increased with the increase of the concentration of TFFC. Compared with AngⅡgroup (20.20 ± 0.83) μmol / L, the difference of NO concentration in CFs culture supernatant was (23.23 ± 0.70) μmol / L after 72 h intervention with 100 mg / L TFFC ). The activity of NOS in culture supernatant was (20.43 ± 0.53) U / L, which was also significantly different from that in AngⅡ group (20.90 ± 0.85) U / L (P <0.01). There was a significant positive correlation between NO concentration and NOS activity (r = 0.964, P <0.01). CONCLUSION: TFFC can inhibit the cell cycle arrest induced by AngⅡ in CFs induced by AngⅡ, and its effect may be related to the up-regulation of NO-NOS activity.