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[摘要]
目的 观察广枣总黄酮(TFCF)对血管紧张素II(Ang II)诱导心脏成纤维细胞(CFs)胶原合成的影响。方法 采用差速贴壁法培养新生大鼠CFs,在体外建立Ang II诱导CFs增殖模型。采用3H-脯氨酸掺入法及羟脯氨酸比色法检测CFs胶原合成,分别观察一氧化氮合酶抑制剂(L-NAME)、鸟苷酸环化酶抑制剂(ODQ)及TFCF对Ang II诱导CFs胶原合成的影响;采用硝酸还原酶法测定细胞培养液中一氧化氮(NO)水平;化学比色法测细胞上清液中一氧化氮合酶(NOS)水平;放免法测定细胞内环鸟苷酸(cGMP)水平。结果 TFCF 25~100 mg/L呈剂量依赖性抑制Ang II诱导的CFs胶原合成,但这种作用可被L-NAME及ODQ部分阻断;TFCF作用细胞后NO、NOS、cGMP水平升高。结论 TFCF在一定质量浓度范围对Ang II诱导的CFs胶原合成有抑制作用,NO-cGMP信号通路可能是其发挥作用的途径之一。
[Key word]
[Abstract]
Objective To investigate the effects of total flavones from Chorspondiatis Fructus (TFCF) on collagen synthesis in cardiac fibroblasts (CFs) induced by angiotensin II (Ang II). Methods The CFs of rats were cultured by the differential adherence method. CFs proliferation model was established by Ang II in vitro. The collagen synthesis was measured by 3H-proline incorporation assay and hydroxyproline chromatography in order to observe the effects of TFCF, L-NAME, and ODQ on collagen synthesis induced by Ang II. The levels of NO, NOS, and cGMP in culture media were measured by nitrate reductase method, chemical colorimetric method, and radioimmunoassay, respectively. Results TFCF (25—100 mg/L) inhibited collagen synthesis of CFs and these effects were blocked by pretreatment with L-NAME or ODQ. Pretreatment of CFs with TFCF resulted in the increase of NO, NOS, and cGMP levels. Conclusion TFCF could inhibit the collagen synthesis of CFs induced by Ang II in a dose dependent manner and the inhibitory effects might be partly correlated with NO-cGMP signaling pathways.
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[基金项目]
国家自然科学基金资助项目(81060362);内蒙古自治区自然科学基金资助项目(2009BS1204)