摘要
目的研究萘哌地尔对大鼠血管平滑肌细胞增殖的抑制作用,并对作用机制进行探讨。方法培养大鼠血管平滑肌细胞,并用去甲肾上腺素诱导其进行增殖,用细胞计数法检测和比色法研究萘哌地尔对大鼠血管平滑肌细胞增殖的影响;用应荧光法测定细胞内的钙浓度,用实时PCR对相关基因的表达进行检测。结果培养的大鼠胸主动脉平滑肌细胞,经鉴定阳性数量达到96%,符合实验标准。各组细胞数差异无统计学意义,且加药前后细胞的形态没有明显变化;与对照组比较,去甲肾上腺素(NE)能明显诱导细胞的增殖,吸光度(OD)值为(0.55±0.016),显著高于对照组的(0.42±0.020),差异有统计学意义(P<0.05);萘哌地尔可抑制细胞的增殖,OD值为(0.30±0.19),与对照组比较,差异有统计学意义(P<0.05);与对照组比较,NE能显著促进细胞中c-myc和c-fosmRNA的表达,分别为(132.5±8.9)与(670.5±9.6),显著高于对照组的(81.2±2.6)与(22.8±1.6),差异有统计学意义(P<0.05);对HRG-1 mRNE的表达具有显著抑制作用;萘哌地尔对NE诱导的c-fosmRNA和c-myc过度表达和HRG-1 mRNA表达的降低具有明显的抑制作用;钙离子浓度与基因表达研究结果一致。结论萘哌地尔可明显的抑制大鼠血管平滑肌细胞增殖,其作用机制可能为降低细胞内血钙浓度,调节相关基因的表达。
Objective To study the inhibition effect of naftopidil on the proliferation of rat vascular smooth muscle cells, and to discuss its mechanisms. Methods The rat vascular smooth muscle cells were cultured and proliferated by norepinephrine （ NE ） inducing. The ef-fect of naftopidil on the proliferation of rat vascular smooth muscle cells was detected by using the colorimetric method and the cell counting method;the intracellular calcium concentration was determined by the fluorescence method and the expression of related genes was detected by the real-time PCR. Results The identified positive number of cultured rat aortic smooth muscle cells reached 96%and conformed to the experimental criteria. No statistically significant differences in cell number among various groups existed, moreover no obvious changes of the cell morphology were found before and after adding medication;compared with the control group, NE could significantly induce the cell proliferation, its OD value （ 0. 55 ± 0. 016 ） was significantly higher than that in control group （ 0. 42 ± 0. 020 ） , the difference was statistically significant （ P〈0. 05 ）; naftopidil could inhibit the cell proliferation, the OD value was 0. 30 ± 0. 19, the difference was statistically significant compared with the control group （ P〈0. 05 ）;compared with the control group, NE could significantly promote the expression of c-myc and c-fosmRNA in cells, which were 132. 5 ± 8. 9 and 670. 5 ± 9. 6, and significantly higher than 81. 2 ± 2. 6 and 22. 8 ± 1. 6 in the control group respectively, the difference was statistically significant （ P〈0. 05 ） , which had significant inhibition effect on the expression of HRG-1 mRNA;naftopidil had significant inhibition effect on the over expression of NE induced c-fosmRNA and c-myc and the decrease of HRG-1 mRNA expression;the calcium ion concentration was consistent to the results of gene expression. Conclusion Naftopidil can significantly inhibit the proliferation of rat vascular smooth muscle cel
出处
《中国药业》
CAS
2014年第17期24-25,共2页
China Pharmaceuticals
关键词
萘哌地尔
大鼠
血管平滑肌
细胞
增殖
naftopidil
rat
vascular smooth muscle
cell
proliferation
