【摘要】 目的研究血管平滑肌细胞(VSMCs)增殖过程中钙调神经磷酸酶(CaN)对三磷酸肌醇受体Ⅰ型(IP3RⅠ)表达的调节。方法植块法分离培养VSMCs,比色法(MTT法)测定VSMCs的增殖,荧光染色法测定VSMCs的活性,RT-PCR检测IP3RⅠ型mRNA转录水平,免疫印迹法(Westernblot)测定IP3RⅠ型蛋白质翻译水平。结果苯肾上腺素(PE)组VSMCs吸光度显著高于对照组(P<0·05),环孢素A(CsA)组和CsA+PE组VSMCs吸光度显著低于对照组(P<0·05)。PE组活细胞百分数与对照组无显著差异(P>0·05),CsA组和CsA+PE组活细胞百分数显著低于对照组(P<0·05)。PE组IP3RⅠ型mRNA表达水平显著高于对照组(P<0·05),CsA组和CsA+PE组mRNA水平显著低于对照组(P<0·05)。PE组IP3RⅠ型蛋白质表达水平显著高于对照组(P<0·05),CsA组和CsA+PE组蛋白质表达水平显著低于对照组(P<0·05)。结论CaN调节VSMCs增殖与活性,并且在转录水平和翻译水平调节IP3RⅠ的表达。更多还原

【Abstract】 Objective To explore the modification of inositol 1, 4, 5-trisphosphate receptor (IP3R) type Ⅰ expression by calcineurin in proliferation of vascular smooth muscle cells (VSMCs). Methods VSMCs were derived from Wistar rats. Cellular growth was studied by using MTT assay and cell viability by acridine orange and ethidium bromide nuclear fluorescent staining. IP3R type Ⅰ expression was determined by RT-PCR and immunoblotting methods. Results The absorbance value was increased in the presence of PE, and lower in the presence of CsA or CsA+PE than that in control (P<0.05). The percentage of viable cells was of no difference between PE group and control (P>0.05), but was decreased in the presence of CsA or CsA+PE (P<0.05). PE accentuated the expression levels of mRNA IP3R type Ⅰ(1.68±0.39 vs control:0.83±0.19, P<0.05), which was attenuated by CsA or CsA+PE ( 0.39±0.12 and 0.51±0.05 vs control:0.83±0.19, P of all <0.05). PE also enhanced the expression levels of protein of IP3R type Ⅰ( 1.78±0.27 vs 1.00±0.00, P<0.05), however CsA or CsA+PE inhibited the increases (0.46±0.37 vs 1.00±0.00; 0.63±0.25 vs 1.00±0.00, P of all<0.05). Conclusion Calcineurin regulates VSMC proliferation and viability. The expression of IP3R type Ⅰ mRNA and protein which was regulated by calcineurin at the transcriptional and translational level.更多还原