RepSox对X射线辐照后血管内皮细胞ALK5表达的抑制作用

Inhibition effect of RepSox on expression of ALK5 in vascular endothelial cells after X-ray irradiation

目的研究X射线辐照后血管内皮细胞中激活素受体样激酶5(ALK5)的表达变化以及RepSox对ALK5及其下游靶点的抑制作用。方法将人脐静脉内皮细胞(HUVEC)分为对照组(Control)、辐照组(IR)和RepSox干预组(IR+RepSox)。RepSox干预组辐照前24 h进行干预,然后用X射线进行照射。辐照后24 h采用CCK-8法检测细胞增殖,qRT-PCR检测细胞ALK5、Smad2和Smad3 mRNA表达,Western blot检测ALK5、p-Smad2/3蛋白表达。结果辐照后24 h,与对照组比较,辐照组细胞数量下降(P<0.05);ALK5、Smad2、Smad3 mRNA表达均上调(P<0.05),ALK5、p-Smad2/3蛋白表达均上调(P<0.05)。与辐照组比较,RepSox干预组细胞数量升高(P<0.05);ALK5、Smad2、Smad3 mRNA表达均下调(P<0.05),ALK5、p-Smad2/3蛋白表达均下调(P<0.05)。结论辐照可以导致血管内皮细胞ALK5及其下游基因表达上调,使用RepSox可以有效地抑制ALK5及其下游基因的表达,从而降低辐照对血管内皮细胞的损伤。

Objective To investigate the changes of activin receptor-like kinase 5(ALK5)expression in vascular endothelial cells after X-ray irradiation and the inhibitory effect of RepSox on ALK5 and its downstream targets.Methods Human umbilical vein endothe-lial cells(HUVECs)were divided into control group,irradiation group and RepSox intervention group(irradiation+RepSox).HUVECs in RepSox intervention group were interfered 24 h before X-ray irradiation.Cell proliferation was detected at 24 h after irradiation by CCK-8 method,the mRNA levels of ALK5,Smad2 and Smad3 were detected by qRT-PCR,and the protein expression of ALK5 and p-Smad2/3 was detected by Western blot.Results Compared with control group,the number of cells was decreased in irradiation group 24 h after irradiation(P<0.05);the mRNA levels of ALK5,Smad2 and Smad3 were up-regulated in irradiation group(P<0.05),and the protein expression of ALK5 and p-Smad2/3 was up-regulated(P<0.05).Compared with irradiation group,the number of cells was increased in RepSox intervention group(P<0.05);the mRNA levels of ALK5,Smad2 and Smad3 were down-regulated(P<0.05),and the protein expression of ALK5 and p-Smad2/3 was down-regulated(P<0.05).Conclusion Irradiation can lead to the upregulation of ALK5 and its downstream gene expression in vascular endothelial cells,and RepSox can effectively inhibit the expression of ALK5 and its downstream genes,thus reducing the damage of irradiation on vascular endothelial cells.