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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)。结论 C
Objective To investigate the regulation of calcineurin (CaN) signaling pathway on arginine vasopressin (AVP) -induced proliferation of neonatal rat cardiac fibroblasts (CFs). Methods The CFs of neonatal Sprague-Dawley rats were cultured by trypsin digestion. The cell number was determined by MTT colorimetric assay. The cell cycle was analyzed by flow cytometry. The activity of CaN was measured by spectrophotometer. Results (1) The A490 value of MTT colorimetric absorbance of CFs increased with the increase of AVP concentration. The A490 values of 10 -7 mol / L AVP and 10 -6 mol / L AVP groups (0 17 ± 0 0 1 and 0 0 18 ± 0 0 1) was significantly different from that of the blank control group (0 11 ± 0 0 1) (P <0.01). The A490 values of AVP + CsA group were all lower than those of the corresponding AVP group, Compared with 10 -7 mol / L AVP group and 10 -6 mol / L AVP + CsA group, there was a significant decrease in 10 -7 mol / L AVP and 10 -6 mol / L AVP + CsA groups compared with 0 7 ± 0 0 1 and 0 15 ± 0 0 1 group Statistical significance (P <0.01). (2) The percentage of cell cycle S phase of CFs treated with 10 -7 mol / L AVP was 17.86 ± 3.18, which was significantly higher than that of the blank control group (12.10 ± 2.38) (P <0.01 ). The percentage of S phase of CFs in 10 -7 mol / L AVP + CsA group (13 76 ± 2 5 2) was significantly different from that in 10 -7 mol / L AVP group (P 0 05). (3) The CaN activity of CFs in 10 -7 mol / L AVP group was 0 30 ± 0 0 6 kU / mg, which was significantly different from the control group (0 14 ± 0 0 3 kU / mg) (P 0 01). Conclusion