摘要
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.
Abstract
Objective:To evaluate the effect of total flavones of fructus chorspondiatis (TFFC) on the cell cycle of cardiac fibroblasts (CFs) isolated from Sprague-Dawley rat induced by angiotensin Ⅱ (Ang Ⅱ),and its relationship with activity of NOS-NO (nitric oxygen syntheses-nitric oxygen) system.Methods:CFs were cultured in vitro and stimulated with 10-7 mol/L Ang Ⅱ.The cell cycle was determined by flow cytometry (FCM).The activity of the actate dehydrogenase (LDH) was used to detect cardiac fibroblasts membrane permeability changes.The cellular growth and proliferation were detected by using trypan blue staining.Nitrate enzyme reverting method was also adopted to evaluate the NO content,and NOS activity was estimated by chemical colorimetric method.Results:Based on the analysis of FCM cell cycle,it showed that the percentage of cells on S and PI stage in CFs were gradually declined as TFFC concentration increased,while the percentage of cells on G0/G1 stage raised (P＜0.01).NO concentration in supernatant [(23.23 ± 0.70) μ mol/L] treated by 100 mg/L TFFC for 72 hours,and was significantly higher than that of Ang Ⅱ group[(20.20 ± 0.83) μ mol/L,P＜0.01].NOS activity in supernatant[(20.43 ± 0.53) U/L] treated by 100 mg/L TFFC for 72 hours,and was also significantly higher than that of Ang Ⅱ group[(20.90 ± 0.85) U/L,P＜0.01).There was a significant positive relevance between NO contents and NOS activity(r=0.964,P＜0.01).Conclusion:TFFC can inhibit the cell cycle proliferation of CFs in vitro,which maybe correlates with upregulation of NOS-NO system.
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维普
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