米诺地尔对血管平滑肌细胞迁移及高血压相关基因离子转换调节器5表达的影响

Effects of minoxidil on migration of vascular smooth muscle cells and expression of hypertension related gene FXYD domain containing ion transport regulator 5

目的 探究米诺地尔对血管平滑肌细胞迁移及高血压相关基因离子转换调节器5(FXYD5)表达的影响。方法 体外培养人主动脉血管平滑肌细胞(HASMC),并分为空白组(正常培养HASMC细胞)及低、中、高剂量实验组(分别加入含终浓度为0.1,0.5和1.0 mmol·L^(-1)米诺地尔溶液的不含胎牛血清、含90%高糖的DMEM培养基)。用划痕实验检测各组HASMC细胞的迁移能力,用蛋白质印迹法检测FXYD5、N-钙黏附蛋白(N-cadherin)、Ras同族体基因家族成员A(RhoA)和Rho激酶1(ROCK1)蛋白的表达情况。结果 中、高剂量实验组和空白组的划痕愈合率分别为(45.96±6.85)%,(30.15±4.38)%和(78.94±9.76)%,FXYD5蛋白相对表达水平分别为0.53±0.08,0.16±0.03和1.13±0.16,N-Cadherin蛋白相对表达水平分别为0.45±0.08,0.20±0.03和0.98±0.15,RhoA蛋白相对表达水平分别为0.47±0.08,0.20±0.03和1.06±0.16,ROCK1蛋白相对表达水平分别为0.77±0.12,0.36±0.05和1.25±0.19。中、高剂量实验组的上述指标与空白组比较,差异均有统计学意义(均P<0.05)。结论 米诺地尔可通过下调FXYD5表达,抑制RhoA/ROCK通路活化,从而实现对血管平滑肌细胞增殖和迁移的抑制作用。

Objective To investigate the effect of minoxidil on the migration of vascular smooth muscle cell and the expression of hypertension-related gene FXYD domain containing ion transport regulator 5(FXYD5). Methods Human aortic vascular smooth muscle cells(HASMC) were cultured in vitro and divided into blank group(normal culture HASMC cells) and experimental-L,-M,-H groups(DMEM medium without fetal bovine serum and containing 90% high glucose with final concentrations of 0.1, 0.5 and 1.0 mmol·L^(-1)minoxidil solution). The migration ability of HASMC cells in each group was detected by scratch test. The protein expressions of FXYD5, N-cadherin, Ras homolog gene family member A(RhoA) and Rho kinase1(ROCK1) were detected by Western blotting. Results The wound healing rates of HASMC cells in experimental-M,-H groups and blank group were(45.96±6.85)%,(30.15±4.38)% and(78.94±9.76)%;the expression levels of FXYD5 protein were 0.53±0.08, 0.16±0.03 and 1.13±0.16;the expression levels of N-Cadherin protein were 0. 45 ± 0. 08, 0. 20 ± 0. 03 and 0. 98 ± 0. 15;the expression levels of RhoA protein were0. 47 ± 0. 08, 0. 20 ± 0. 03 and 1. 06 ± 0. 16;the expression levels of ROCK1 were 0. 77 ± 0. 12, 0. 36 ± 0. 05 and1. 25 ± 0. 19, respectively. Compared with blank group, the above indexes of experimental-M,-H groups werestatistically significant(allP< 0. 05).Conclusion Minoxidil can inhibit the proliferation and migration of vascularsmooth muscle cells by down-regulating the expression of FXYD5 and inhibiting the activation of RhoA/ROCK pathway.