钙调神经磷酸酶调控血管升压素诱导大鼠心脏成纤维细胞胶原合成的作用

Calcineurin mediates the collagen synthesis of rat cardiac fibroblasts induced by arginine vasopressin

目的:探讨钙调神经磷酸酶(CaN)信号通路对血管升压素(AVP)诱导新生大鼠心脏成纤维细胞(CF)胶原合成的调控作用。方法:采用胰酶消化法培养新生SD大鼠CF,应用羟基脯氨酸比色法测定细胞培养上清中羟基脯氨酸含量,CaN活性通过分光光度计测定。结果:①CF培养上清中羟基脯氨酸含量随着AVP浓度的升高而增加,其中10-7mol/L AVP和10-6mol/L AVP组羟基脯氨酸含量与空白对照组比较有显著性差异(均P<0.01);在0.5μg/m l CaN的特异性抑制剂环孢素共同干预下,羟基脯氨酸含量明显减少,并有统计学意义(P<0.01)。②随着AVP浓度的升高,CF内CaN活性亦随之增高,与空白对照组比较,10-7和10-6mol/L AVP组细胞内CaN活性显著增高(P<0.01)。③在10-7mol/L AVP作用下,随着细胞内CaN活性的升高,CF培养上清中羟基脯氨酸含量随之增加,两者之间呈显著正相关(r=0.91,P<0.05)。④在10-7mol/L V1受体拮抗剂[d(CH2)5-Tyr2(M e)]AVP和10-7mol/L AVP共同作用下,CF中CaN活性和培养上清中羟基脯氨酸含量与单独10-7mol/LAVP作用组比较均显著降低,并有统计学意义(均P<0.01)。结论:CaN信号通路在AVP诱导CF胶原合成过程中起到了重要的调控作用,该通路的激活可能是通过V1受体介导的。

Objective ：To investigate the effects of calcineurin signaling pathway on the collagen synthesis of rat cardiac fibroblasts （CF） induced by arginine vasopressin （AVP）. Methods：CF of neonatal Sprague-Dawley rats were secured by trypsinization and Cultured. Collagen content of CF culture medium was estimated by hydroxyproline colorimetry. Calcineurin activity was determined with spectrophotometer. Results ：① AVP increased the content of hydroxyproline in culture medium of CF in a concentration dependent manner. But in the presence of 0.5 μg/ml cyclosporin A （CsA）, the specific inhibitor of calcineurin, the hydroxyproline content of culture medium in corresponding AVP ＋ CsA group was significantly lower than that of 10^-7 mol/L AVP or 10^-6mol/L AVP group （both P 〈0.01 ）. ② The calcineurin activity was gradually elevated with the increase of AVP concentration. Compared with the control, the activity in 10^-7 mol/L AVP and 10^-6mol/L AVP group significantly increased （P 〈 0.01 ）. ③ The correlation analysis indicated there was a positive correlation between the calcineurin activity and the content of hydroxyproline in culture medium of CF （r = 0.91, P 〈 0.05）. ④ In the presence of [ d （CH2 ） 5-Tyr^2 （Me） ] AVP, the antagonist of V1 receptor, the elevation of ealeineurin activity stimulated by AVP was completely inhibited （P 〈0.01 ） and the increase of hydroxyproline content was blocked too（P 〈0.01 ）. Conclusion：Above results suggest that ealeineurin signaling pathway probably plays a great role in collagen synthesis of CF induced by AVP, which might be mediated through V1 receptor.