摘要
目的 探讨钙调神经磷酸酶 (CaN)信号通路对精氨酸升压素 (AVP)诱导新生大鼠心脏成纤维细胞 (CFs)增殖的调控作用。方法 采用胰酶消化法培养新生Sprague-Dawley大鼠CFs,MTT比色法测定细胞数目 ,应用流式细胞仪分析细胞周期 ,CaN活性通过分光光度计测定。结果 (1)CFs的MTT比色法吸光度A490 值随着AVP作用浓度的升高而增加 ,其中10 -7mol/LAVP和 10 -6mol/LAVP组A490 值(分别为 0 17±0 0 1和 0 18± 0 0 1)与空白对照组 (0 11± 0 0 1)比较有显著差异 (P <0 0 1) ;不同浓度的AVP +CsA组A490 值均较相应的AVP组降低 ,其中 10 -7mol/LAVP +CsA组和 10 -6mol/LAVP +CsA组分别为 0 13± 0 0 1和 0 15± 0 0 1,与 10 -7mol/LAVP和 10 -6mol/LAVP组比较显著降低 ,并有统计学意义 (P <0 0 1)。(2 ) 10 -7mol/LAVP作用下CFs细胞周期S期百分率为 17.86± 3.18,与空白对照组 (12 .10± 2 .38)比较明显升高 ,并有统计学意义(P <0 0 1)。 10 -7mol/LAVP +CsA组CFs细胞周期S期百分率 (13 76± 2 5 2 )与 10 -7mol /LAVP组比较有显著差异(P <0 0 5 )。 (3) 10 -7mol/LAVP组CFs的CaN活性为 0 30± 0 0 6kU/mg,与对照组 (0 14± 0 0 3kU/mg)比较差异显著(P <0 0 1)。
Objective To investigate the effects of calcineurin signaling pathway on the proliferation of rat myocordial fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods Cultured CFs of neonatal SD rats were isolated by trypsinization. Cell number and cell cycle were evaluated by MTT assay and flowcytometry technique. Calcineurin activity was determined with spectrophotometer. Results ①AVP increased the absorbance at 490 nmol (A_ 490 ) of CFs in a concentration dependent manner. In comparison with A_ 490 of the control group(0.11±0.01),the A_ 490 value in 10 -7 mol/L AVP and 10 -6 mol/L AVP group were markedly increased(0.17±0.01,0.18±0.01,P <0.01 respectively). In the presence of 0.5 μg/mL Cyclosporin A(CsA),the specific inhibitor of calcineurin,A_ 490 value of CFs in AVP+CsA group (0.13±0.01,0.15±0.01 respectively ) were significantly lower than that of 10 -7 mol/L AVP and 10 -6 mol/L AVP group (both P <0.01). ② Compared with the control(12.10±2.38),cells in S stage were increased in 10 -7 mol/L AVP group(17.86±3.18,P <0.01),but in the presence of 0.5 μg/mL CsA,the percentage of S stage cells was decreased (13.76±2.52,P <0.05). ③Calcineurin activity in 10 -7 mol/L AVP group was higher than that of control group (0.30±0.06 vs 0.14±0.03 kU/mg, P <0.01). Conclusion These results indicate AVP induces the activation of calcineurin which was in hibited by CsA. Calcineurin signaling pathway may play a role in CFs proliferation induced by AVP.
出处
《高血压杂志》
CSCD
2004年第1期58-61,共4页
Chinese Journal of Hypertension
基金
陕西省自然科学基金资助项目 (2 0 0 0SM2 6)
