白细胞介素10对血管升压素诱导心脏成纤维细胞增殖及胶原合成的影响

Effects of Interleukin-10 on the Proliferation and Collagen Synthesis Induced by Arginine Vasopressin

目的研究白细胞介素10(IL-10)对血管升压素(AVP)诱导大鼠心脏成纤维细胞(CFs)增殖及Ⅰ、Ⅲ型胶原合成的影响。方法以培养的新生SD大鼠CFs为实验模型,四氮唑盐(MTT)比色法检测CFs增殖,流式细胞仪技术(FCM)测定细胞周期,逆转录聚合酶链式反应(RT-PCR)检测Ⅰ、Ⅲ型胶原mRNA表达水平。结果(1)10-7mol/L AVP作用24h,CFs的吸光度(A490)为0·216±0·013,较对照组(0·132±0·006)显著增加(P<0·01)。(2)IL-10呈浓度依赖性下调AVP诱导的CFs的A490的增加,其10-8g/mLIL-10干预组CFs的A490(0·157±0·029)较AVP组显著降低(P<0·01)。(3)AVP组CFs的S期百分率及增殖指数(12·30±0·71,19·58±0·88)较对照组(4·22±0·48,7·12±0·62)显著增加(P<0·01);而10-9g/mLIL-10干预组,CFs的S期百分率及增殖指数(9·56±1·13,13·86±1·28)较AVP组显著降低(P<0·01)。(4)AVP组Ⅰ、Ⅲ型胶原mRNA表达水平(1·45±0·06,1·06±0·06)较对照组(1·03±0·05,0·77±0·05)显著增加(P<0·01)。而IL-10可浓度依赖性的下调AVP诱导的CFs的Ⅰ、Ⅲ型胶原mRNA表达,其10-8g/mL IL-10干预组Ⅰ、Ⅲ型胶原mRNA表达水平(1·14±0·06,0·88±0·02)显著低于AVP组(P<0·01)。结论IL-10具有抑制AVP诱导CFs增殖和胶原合成的作用,这可能对预防和逆转心脏重构有一定的价值。

Objective To study the effect of interleukin-10 （IL-10） on the proliferation and collagen synthesis induced by arginine vasopressin（AVP） in cultured cardiac fibroblasts（CFs）. Methods The CFs of neonatal Sprague-Dawley rats were isolated and cultured. Cell number were determined by MTT assay （A490 value）. Cell cycle distribution was assessed using flowcytometry technique. Ⅰ-type and Ⅲ-type collagen mRNA in CFs were analyzed by the reverse transcriptase-polymerase chain reaction （RT-PCR）. Results （1）10^-7 mol/L AVP significantly increased A490 value of CFs（0. 216±0. 013 vs control： 0. 132±0. 006, P〈20.01）. （2） IL-10 attenuated the A490 value in a concentration dependent manner. （3）The percentage of the cells in S stage（12. 30±0.71 vs 4.22±0.48, P 〈0.01） and proliferation index（PI 19.58 ±0.88 vs 7. 12±0.62, P 〈20.01） were markedly increased in AVP goup compared with the control. （4） The mRNA abundances of Ⅰ type（1.45±0.06 vs 1. 03±0. 05, P〈0. 01）and Ⅲ type collagen stimulated by AVP（ 1. 06±0.06 vs 0. 84±0.05, P〈0. 01 ） were significantly higher than that of the control. IL-10 attenuated the mRNA abundances of Ⅰ and Ⅲ type collagen of AVP group in a concentration dependent manner. Conclusion IL-10 effectively inhibits the cell proliferation and collagen synthesis of CFs induced by AVP, suggesting IL-10 may play a role in the regression of heart remolding.