摘要
目的 :观察精氨酸加压素 (AVP)作用下 p2 7蛋白表达在心脏成纤维细胞 (CFs)中的变化 ,探讨 p2 7蛋白调控AVP促 CFs增殖的作用。方法 :以培养的新生 SD大鼠 CFs为实验动物模型 ,采用胰酶消化、差速贴壁法培养CFs,四氮唑盐 (MTT)比色法检测细胞增殖 ,碘化丙啶标记细胞 DNA ,p2 7蛋白的单抗和标记了 FITC的二抗标记细胞内的 p2 7蛋白 ,采用流式细胞分析仪技术测定细胞周期及 p2 7蛋白表达的阳性率。结果 :1随 AVP浓度的增加 ,MTT法测得的 CFs的 A值呈递增趋势 ,其中 10 - 7,10 - 6 m ol/ L AVP组分别为 0 .30 7± 0 .0 0 9,0 .337± 0 .0 0 7,均明显地高于对照组 (0 .2 98± 0 .0 0 9) ,并有统计学意义 (分别 P<0 .0 5 ,P<0 .0 1) ;2随 AVP浓度的增加 ,CFs的 S期细胞百分率和细胞增殖指数 (PI)逐渐增加 ,G0 / G1 期百分率下降 ,10 - 7,10 - 6 mol/ L AVP组与对照组比较差异非常显著 (P<0 .0 1) ;3CFs的 p2 7蛋白表达阳性率随 AVP浓度的增加而呈递减趋势 ,其中 10 - 7,10 - 6 m ol/ L AVP组 (分别为 71.6 %± 2 .2 % ,6 3.1%± 2 .3% )明显低于对照组 (86 .5 %± 2 .3% ) ,并有统计学意义 (P<0 .0 1)。结论 :p2 7蛋白是 AVP促 CFs增殖过程的重要负性调节因子 ,AVP通过下调 p2 7蛋白发挥促 CFs增殖作用 。
AIM: To study the change of p27 expression in cardiac fibroblasts (CFs) proliferation, investigate the role of p27 in the CFs proliferation modulated by arginine vasopressin(AVP). METHODS: As a model, neonatal rat CFs was cultured with enzymatic dissociation. AVP in different concentration was added. Proliferation was measured through MTT assay. To identify the cell cycle,propidium iodide was used to lable DNA. Cell cycle distribution and p27 expression were determined with flow cytometre. RESULTS: AVP increased A value of CFs in a concentration dependent manner. The value of 10 -7 and 10 -6 mol/L AVP groups by MTT assay were 0.307±0.009 and 0.337±0.007 respectively, which were significantly higher than that of the control (0.298±0.009)( P <0.05, P <0.01,respectively). The percentage of CFs in S stage and proliferation index (PI) were increased significantly with the elevation of AVP concentration,concomitant with a significant decrease in the percentage of G 0/G 1 stage cells. Positive rate of p27 expression in CFs was downregulated by AVP also in a concentration dependent manner. Positive rate of p27 expression in CFs of 10 -7 and 10 -6 mol/L AVP groups (71.6%±2.2%,63.1%±2.3%) were significantly lower than that of the control (86.5%±2.3%)( P <0.01). CONCLUSION:p27 may be a negative modulating factor in CFs proliferation .By downregulating p27 expression,AVP can increase CFs proliferation,thus,participate in the pathogenesis and development of cardiac fibrosis.
出处
《心脏杂志》
CAS
2001年第2期122-124,共3页
Chinese Heart Journal
