替米沙坦对人脐带动脉平滑肌细胞增殖的抑制作用

The inhibition of telmisartan on human umbilical artery smooth muscle cells proliferation

目的研究替米沙坦对体外培养的人脐带动脉血管平滑肌细胞(HUASMCs)增殖的抑制作用及其机制。方法组织块贴壁法原代培养HUASMCs,选传至3代的细胞实验;MTT比色法检测替米沙坦的同一浓度(97.16μmol·L^(-1))在不同作用时间(3~96 h)和不同浓度(0.97~971.59μmol·L^(-1))在同一作用时间(48 h)对HUASMCs增殖的抑制作用;RT-PCR法测定3种浓度的替米沙坦(1.94、19.43、194.32μmol·L^(-1))对HUASMCs表达血管内皮细胞生长因子(VEGF)mRNA的影响;ELISA法测定上述3种浓度的替米沙坦对HUASMCs分泌VEGF水平的影响。结果替米沙坦的作用时间在3~48 h范围内,对HUASMCs增殖的抑制作用呈时间依赖性(P<0.05),48 h抑制作用最强,抑制率为(29.12±2.89)%,72 h、96 h与48 h比较,抑制作用无统计学意义(P>0.05)。替米沙坦的作用浓度在0.97~194.32μmol·L^(-1)范围内,对HUASMCs增殖的抑制作用呈浓度依赖性(P<0.05),作用浓度为194.32μmol·L^(-1)时抑制作用最强,抑制率为(41.46±3.36)%,388.64μmol·L^(-1)、582.95μmol·L^(-1)、971.59μmol·L^(-1)与194.32μmol·L^(-1)比较,抑制作用无统计学意义(P>0.05);高(194.32μmol·L^(-1))、中(19.43μmol·L^(-1))、低(1.94μmol·L^(-1))3种浓度的替米沙坦对HUASMCs表达VEGF的mRNA和因子均有抑制作用(P<0.05),并呈浓度依赖性(P<0.05)。结论替米沙坦可抑制体外培养的HUASMCs增殖,使其表达的VEGF下调。

Objective To investigate the inhibition and mechanism in human umbilical artery smooth muscle cells （HUASMCs） Proliferation of telmisartan. Methods HUASMCs were primarily cultured with tissue adherent method and the third generation cells were selected for immunohistochemical staining. The inhibition of telmisartan （97. 16 μmol·L^-1 ） at different time （3 -96 h） and different concentrations （0.97 -971.59 μmol·L^-1） at the same time （48 h） for the proliferation of HUASMCs were determined by MTT assay. The effect of telmis- artan （ 1.94, 19.43, 194. 32μmol·L^-1 ） on expression of vascular endothelial growth factor VEGF mRNA in HUASMCs was determined by RT-PCR. The effect of telmisartan （1.94, 19.43, 194. 32μmol·L^-1） on secretion level of VEGF in HUASMCs was determined by double antibody sandwich ELISA. Results Telmisartan inhibited the activity of HUASMCs in every detection time point （P 〈 0. 05 ） and the inhibition rate of 3 to 48 h increased with the time and reached its peak after 48 h, inhibition rate reached （29.12 ±2. 89 ）% , the inhibition of proliferation at 72 h and 96 h compared with 48 h, the difference had no statistical significance （P 〉0. 05）. When telmisartan concentration at the range of 0. 97 - 194. 32μmol·L^-1 the proliferation inhibition on HUASMCs presented dosage dependent （ P 〈 0, 05 ）. When telmisartan concentration in 194.32 μmol·L^-1, the inhibitory effect was strongest, inhibition rate was （41.46 ± 3.36）%. With the concentration was expanding （388.64 μmol·L^-1 -971.59μmol·L^-1 ） , the inhibition of proliferation had no significant difference compared with 194. 32 μmol·L^-1 （ p 〉 0. 05 ）. Concentrations of telmisartan with high, medium, low （ 1.94, 19.43, 194.32μmol·L^-1） inhibited the expression of VEGF mRNA of HUASMCs （P 〈 0. 05）. Three kinds of concentrations of telmisartan inhibited the secretion of VEGF of HUASMCs （P 〈 0. 05 ） and presented dosage dependent. Conclusion Telmisartan inhibited the vitro proliferation of HUASMCs. Telmisartan inhibited the expression of VEGF mRNA and the secretion of VEGF in HUASMCs.