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目的观察广枣总黄酮(total flavones of fructus chorspondiatis,TFFC)对血管紧张素Ⅱ(angiotensin,AngⅡ)诱导心脏成纤维细胞(cardiac fibroblasts,CFs)增殖的影响。方法采用差速贴壁法培养新生大鼠CFs,在体外建立AngⅡ诱导CFs增殖模型。采用四甲基氮唑盐(methyl thiazole tetrazolium,MTT)比色法及3H-胸腺嘧啶核苷(3H-deoxythy-midine,3H-TdR)掺入法检测细胞增殖,分别观察一氧化氮合酶抑制剂[nitric oxide synthase inhibitor,Nω-nitro-L-arginine methyl ester(L-NAME)]、鸟苷酸环化酶抑制剂(1H-[1,2,4]oxadiazolo[4,3-a]-quinoxalin-1-one,ODQ)及TFFC对AngⅡ诱导CFs增殖的影响;采用硝酸还原酶法测细胞培养液中一氧化氮(nitric oxide,NO)水平;化学比色法测细胞培养液中一氧化氮合酶(nitricoxide synthase,NOS)水平;放免法测定细胞内环磷酸乌苷(cyclic guanosine monophosphate,cGMP)水平。结果广枣总黄酮25~100 mg/L呈剂量依赖性抑制AngⅡ诱导的CFs增殖,但这种作用可被L-NAME及ODQ部分阻断;广枣总黄酮作用细胞后NO、NOS、cGMP水平升高。结论广枣总黄酮在一定浓度范围对AngⅡ诱导的CFs增殖有抑制作用,通过NO-cGMP信号通路可能是其发挥作用的途径之一。
Objective To observe the effect of total flavones of fructus chorspondiatis (TFFC) on the proliferation of cardiac fibroblasts (CFs) induced by angiotensin Ⅱ (Angiotensin Ⅱ). Methods CFs of newborn rats were cultured by differential adherence method. The proliferation of CFs induced by Ang Ⅱ was established in vitro. Cell proliferation was detected by methyl thiazole tetrazolium (MTT) colorimetric assay and 3H-deoxythy-midine (3H-TdR) incorporation assay. Nitric oxide synthase Nitric oxide synthase inhibitor (Nω -nitro-L-arginine methyl ester (L-NAME)], guanylate cyclase inhibitor (1H- [1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ) and TFFC on the proliferation of CFs stimulated by AngⅡ. The level of nitric oxide (NO) was measured by nitric acid reductase method and the level of nitric oxide The level of nitric oxide synthase (NOS) was determined by radioimmunoassay. The levels of intracellular cyclic guanosine monophosphate (cGMP) were determined by radioimmunoassay. Results Total flavonoids from jujube could inhibit the proliferation of CFs induced by AngⅡ in a dose-dependent manner, but this effect could be partially blocked by L-NAME and ODQ. The contents of NO, NOS and cGMP Rise. Conclusion The total flavonoids of jujube can inhibit the proliferation of CFs induced by angiotensin Ⅱ in a certain concentration range, and it may be one of the ways through which NO-cGMP signaling pathway plays a role.