广枣总黄酮抑制Ang Ⅱ诱导大鼠心脏成纤维细胞的增殖及机制  

Inhibitory effects and mechanism of total flavones of fructus Chorspondiatis on the proliferation of cardiac fibroblasts induced by AngⅡ

作　　者：贾磊[1] 金明 杨雨民 包俊萍 高晓慧 舒亮[1] 邢慧慧[1] 

机构地区：[1]内蒙古医科大学研究生学院,内蒙古呼和浩特010050 [2]内蒙古林业总医院,内蒙古牙克石022150 [3]内蒙古自治区中医医院,内蒙古呼和浩特010020

出　　处：《时珍国医国药》2014年第6期1308-1311,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81060362);内蒙古自治区自然科学基金(No.2009BS1204)

摘　　要：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 investigate the effects of total flavones of fructus Chorspondiatis （TFFC） on cardiac fibroblasts （ CFs ） proliferation in cultured rat CFs induced by angiotensin Ⅱ（ AngⅡ ）. Methods The CFs was euhured by the differential attachment method. Cell proliferation model was established by stimulation with AngⅡ. Cell proliferation was measured by MTT assay and 3H -deoxythy- midine（3H -TdR） ,in order to observe the effects of TFFC, L- NAME and ODQ on cell proliferation induced by Ang Ⅱ. Nitric oxide （NO） level in culture medium was measured by Griess reagent. Nitrieoxide synthase （NOS） level in culture medium were measured by chemical colorimetric method. The cyclic guanosine monophosphate（cGMP） level was measured by ra- dioimmunoassay. Results TFFC at the dose from 25 to 100 mgl/L inhibited CFs proliferation, and these effects were blocked by pretreatment with L - NAME or ODQ. Pretreatment of CFs with TFFC resulted in increase NO , NOS and cGMP level. Conclu- sion TFFC inhibited CFs proliferation induced by Ang II in dose dependent manner,the inhibitory effects might be partly correla- ted with NO -eGMP signaling pathways.

关 键 词：广枣总黄酮 心脏成纤维细胞 血管紧张素Ⅱ 细胞增殖 鸟苷酸环化酶 

分 类 号：R285.5[医药卫生—中药学]