IL-10抑制AVP诱导大鼠心脏成纤维细胞CaN活性

目的：研究白细胞介素10（IL-10）对精氨酸血管加压素(AVP)诱导大鼠心脏成纤维细胞(CFs)钙调神经磷酸酶（CaN）活性的影响。方法:用培养的新生SD大鼠CFs，四氮唑盐(MTT)比色法检测CFs增殖，流式细胞仪检测细胞周期，分光光度法测定CaN活性。结果:（1）10-7 mol/L AVP组CFs的吸光度（A490）明显高于对照组（P<0.01）。IL-10呈浓度依赖性下调 AVP诱导的CFs的A490值（P<0.05或P<0.01）。IL-10本身对CFs的增殖无抑制作用。（2）10-7 mol/L AVP组CFs的S期百分率及增殖指数均明显高于对照组（均P<0.01）；10-6 g/L IL-10+10-7 mol/LAVP组S期百分率及增殖指数均明显低于AVP组（P<0.01）。（3）10-7 mol/L AVP组CaN活性明显高于对照组（P<0.01）；10-8、10-7、10-6和10-5g/L +10-7 mol/L AVP组的CaN活性均明显低于AVP组（P<0.01），但仍高于对照组（P<0.01或P<0.05）。IL-10呈浓度依赖性下调 AVP诱导CFs的CaN活性。结论:IL-10具有抑制AVP诱导CFs增殖和下调CFs的CaN活性的作用，这可能对预防和逆转心脏重构有一定的价值。

Effects of interleukin-10 on the calcineurin activity in cultured cardiac fibroblasts induced by arginine vasopressin

AIM: The purpose of the present study was to investigate the effect of interleukin-10 (IL-10) on the proliferation and calcineurin (CaN) activity in cultured cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP).METHODS: The CFs of left ventricle in neonatal Sprague-Dawley rats were isolated and cultured by trypsin digestion and selective plating technique. Then the proliferation rates of cells were determined by using the MTT ［3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide］ assay (A490 value). Cell cycle distribution was determined with flowcytometry technique. The CaN activity was measured by ultra-violet spectrophotography.RESULTS: (1) MTT colorimetry showed that 10-7 mol/L AVP significantly increased A490 value of CFs in comparison with control group (P<0.01). IL-10 attenuated the A490 value of AVP group in a concentration dependent manner. The A490 value of the 10-8, 10-7, 10-6, 10-5 g/L IL-10+10-7 mol/L AVP groups was 0.201±0.007, 0.187±0.006, 0.173±0.010 and 0.157±0.029 respectively, all data significantly lower than those in the presence of AVP alone (P<0.05 or P<0.01). (2) The percentage of the cells in S stage and proliferation index were markedly increased in 10-7mol/L AVP group compared with the control (P<0.01, respectively). In the 10-6 g/L IL-10+10-7 mol/L AVP group, the percentage cells in S stage and proliferation index were significantly lower than those in AVP group (P<0.01, respectively). IL-10 itself had no effect on fibroblast proliferation, but reduced AVP-induced fibroblast proliferation. (3) There was a significantly increase in CaN activity in AVP group compared with control (P<0.01). In the 10-8, 10-7, 10-6 and 10-5 g/L IL-10＋10-7 mol/L AVP groups, the CaN activity was 3.22±0.04, 3.06±0.06, 2.53±0.04 and 2.22±0.04, respectively. IL-10 dose-dependently down-regulated the CaN activity induced by AVP (P<0.01, respectively). However, the CaN activity was still higher in IL-10+AVP group than that in control group (P<0.05 or P<0.01).CONCLUSION: Our data indicate that IL-10 regulates the CaN activity of CFs in the cell proliferation induced by AVP, suggesting that IL-10 plays a role in the regression of cardiac remodeling.