血管钠肽抑制低氧诱导的心肌成纤维细胞的增殖和胶原合成

Vasonatrin peptide inhibits hypoxia-induced proliferation and collagen synthesis in cardiac fibroblasts

目的 观察血管钠肽 (Vasonatrin peptide,VNP)对低氧状态下大鼠心肌成纤维细胞 (CFs)增殖及胶原合成的影响 ,探讨其防治低氧所致心肌间质结构重建的作用机制 .方法 培养新生大鼠 CFs,应用流式细胞仪分析细胞周期 ,以 3H- proline掺入率反映胶原合成速率 .结果 单纯低氧不改变 CFs的细胞周期 ,但增加低浓度血清 (2 5 m L· L- 1 )作用基础上的细胞增殖指数 PI(S+G2 / M) / (S+G2 / M+G0 / G1 )(P<0 .0 1) ,10 - 6 m ol· L- 1的 VNP抑制低氧诱导的 PI增加(P<0 .0 1) .低氧可以增加基础胶原合成 (增加 19.6 % ,P<0 .0 5 ) ,也增加低浓度血清 (2 5 m L· L- 1 )诱导的胶原合成速率 (增加 37.2 % ,P<0 .0 1) . 10 - 8～ 10 - 6 mol· L- 1 的 VNP浓度依赖性抑制低氧诱导的胶原合成速率 (P<0 .0 1) .结论 低氧增加胶原合成以及血清刺激的 CFs的增殖反应 ,VNP抑制上述作用 。

AIM To investigate the influence of vasonatrin peptide (VNP) on hypoxia induced proliferation and collagen synthesis in cultured cardiac fibroblasts (CFs) and the preventive mechanism of VNP in the interstitium remodeling of ischaemic heart diseases due to hypoxia. METHODS Neonatal rat CFs were obtained by enzymatic dissociation. Cell cycle kinetics of CFs was analyzed by flow cytometry. Collagen synthesis rate was determined by measuring 3H proline incorporation rate. RESULTS Hypoxia (20~30 mL·L -1 oxygen) alone for 24 h didn't change the cell cycle of cells cultured on serum free medium, but significantly increased the proliferation index [(S+G 2/M) / (S+G 2/M+G 0/G 1)] of CFs with low concentration serum (25 mL·L -1 ). VNP (10 -6 mol·L -1 ) inhibited the effects of hypoxia on low concentration serum induced proliferation ( P <0.01). Hypoxia increased basal (under nomoxide without serum) and low concentration serum induced collagen syntheses (under nomoxide with 25 mL·L -1 serum) by 19.6% ( P <0.05) and 37.2% ( P <0.01) in CFs, respectively. In a concentrate dependent manner, VNP (10 -8 ~10 -6 mol·L -1 ) decreased the incorporation of 3H proline induced by hypoxia for 24 h ( P <0.01). CONCLUSION Hypoxia might enhance serum induced proliferation in CFs and increase basal and serum induced collagen syntheses. VNP may attenuate myocardial fibrosis induced by hypoxia by inhibiting the above effects.