IL-10抑制AVP诱导大鼠心脏成纤维细胞CaN活性

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

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)10-7mol/LAVP组CFs的吸光度(A490)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A490值(P<0.05或P<0.01)。IL-10本身对CFs的增殖无抑制作用。(2)10-7mol/LAVP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);10-6g/LIL-10+10-7mol/LAVP组S期百分率及增殖指数均明显低于AVP组(P<0.01)。(3)10-7mol/LAVP组CaN活性明显高于对照组(P<0.01);10-8、10-7、10-6和10-5g/L+10-7mol/LAVP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。

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 （A490 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 A490 value of CFs in comparison with control group （ P 〈 0. 01）. IL- 10 attenuated the A490 value of AVP group in a concentration dependent manner. The A490 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.