摘要
目的探讨替米沙坦联合羟苯磺酸钙在自发性高血压中对主动脉的保护作用及作用机制。方法将自发性高血压(SHR)大鼠及其WKY大鼠随机分为(n=6):对照组(WKY)、SHR+NC组(未经处理的SHR大鼠)、SHR+替米沙坦组(替米沙坦治疗的SHR大鼠)、SHR+CAD组(CAD治疗的SHR大鼠)、替米沙坦+CAD组(替米沙坦和CAD联合治疗的SHR大鼠),每组6只。利用苏木精-伊红染色观察治疗前后主动脉血管的变化情况;利用ELISA检测炎症因子的表达水平。利用TargetScan预测miR-19b-3p的下游靶基因,利用双荧光素酶实验验证miR-19b-3p以及下游靶基因之间的结合关系。细胞分组:control组,替米沙坦+NC组,NC+CAD组及替米沙坦+CAD组。随后为了验证替米沙坦与羟苯磺酸钙与miR-19b-3p/SOCS1信号轴的调节关系,利用替米沙坦、羟苯磺酸钙单独处理或者联合对VSMC细胞进行处理。细胞分组:空白组、miR-19b-3p inhibitoror组、miR-19b-3p inhibitor+替米沙坦+NC组、miR-19b-3p inhibitor+NC+CAD组、miR-19b-3p inhibitoror+替米沙坦+CAD组。利用qRT-PCR检测miR-19b-3p和SOCS1的表达水平。从自发性高血压大鼠主动脉中分离血管平滑肌细胞(VSMC),利用CCK-8、Transwell小室实验、qRT-PCR以及Western blot方法验证替米沙坦和羟苯磺酸钙对细胞增殖、迁移表型分型以及炎症因子表达的影响。结果与对照组相比,SHR+NC组大鼠的主动脉管壁厚度明显增厚,管腔内壁降低;经SHR+替米沙坦和SHR+CAD组的SHR大鼠胸主动脉管壁厚度降低,管腔半径增加。ELISA结果显示,与对照组相比,SHR+NC组大鼠血液中IL-1β、TNF-α、IL-6的表达水平显著增加(P<0.05),SHR+替米沙坦、SHR+CAD组、替米沙坦+CAD组的大鼠体内炎症因子的表达水平较SHR+NC组显著降低(P<0.05)。与对照组主动脉中miR-19b-3p的表达水平相比,SHR+NC组显著升高,SHR+替米沙坦和SHR+TAD组显著降低(P<0.05)。qRT-PCR结果显示,与对照组SOCS1的表达水平相比,SHR+NC组显著降低,SHR+替米沙坦、SHR+CAD组、替米沙坦+CAD组较SHR+NC组显著上调(P<0.05)。qRT-PCR结果显示,与control组相比,替米沙坦+NC组、NC+CAD组和替米沙坦+CAD组VSMC细胞中α-SMA、SM22α的表达水平显著增高,OPN的表达水平显著下降(P<0.05)。transwell小室实验结果显示,与control组VSMC细胞的迁移相比,替米沙坦+NC组、NC+CAD组和替米沙坦+CAD组显著降低(P<0.05)。与空白组相比,miR-19b-3p inhibitor组IL-1β、TNF-α的表达水平显著降低,进一步添加替米沙坦与羟苯磺酸钙处理后能增强这一结果(P<0.05)。结论替米沙坦与羟苯磺酸钙联用可以通过调节miR-19b-3p/SOCS1信号轴改善自发性高血压大鼠主动脉损伤,抑制炎症因子的表达。
Objective To investigate the protective effect and mechanism of telmisartan and calcium dobesilate on aorta in rats with spontaneous hypertension.Methods Spontaneously hypertensive(SHR)rats and their WKY rats were randomly divided(n=6)into control group(WKY),SHR group,telmisartan group,calcium oxybenesulfonic acid group and combined treatment group.Hematoxylin-eosin staining was used to observe the changes of aortic vessels before and after treatment.The expression level of inflammatory factors was detected by ELISA.The downstream target genes of miR-19b-3p were predicted by TargetScan,and the binding relationship between miR-19b-3p and downstream target genes was verified by double luciferase assay.The expression levels of miR-19b-3p and SOCS1 were detected by qRT-PCR.Cells were divided into:control group,temesartan+NC group,NC+CAD group and temesartan+CAD group.Subsequently,in order to verify the regulatory relationship between temesartan and calcium hydroxybenzenesulfonate and miR-19b-3p/SOCS1 signaling axis,VSMC cells were treated with temesartan and calcium hydroxybenzenesulfonate alone or in combination.Cell were grouped into:blank group,miR-19b-3p inhibitoror group,miR-19b-3p inhibitor+timisartan+NC group,miR-19b-3p inhibitor+NC+CAD group,miR-19b-3p inhibitoror+timisartan+CAD group.Vascular Smooth Muscle cells(VSMC)were isolated from the aorta of spontaneously hypertensive rats.The effects of telmisartan and calcium isobenesulfate on cell proliferation,migration phenotyping and expression of inflammatory factors were investigated by CCK-8,Transwell laboratory assay,qRT-PCR and WB.Results Compared with the control group,the aortic wall thickness was significantly thickened and the lumen inner wall was decreased in the SHR+NC group;the thickness of the thoracic aortic wall was decreased and the lumen radius was increased in SHR rats via SHR+timisartan and SHR+CAD groups.ELISA results showed that the expression levels of IL-1β,TNF-α,and IL-6 in the blood of rats in the SHR+NC group were significantly increased compared with those in the control group(P<0.05),and the expression levels of inflammatory factors in the rats in the SHR+timisartan,SHR+CAD,and timosartan+CAD groups were significantly decreased compared with those in the SHR+NC group(P<0.05).Compared with the expression level of miR-19b-3p in the aorta of the control group,it was significantly higher in the SHR+NC group and significantly lower in the SHR+timisartan and SHR+TAD groups(P<0.05).The qRTPCR results showed that the expression level of SOCS1 was significantly decreased in the SHR+NC group compared with the control group,and significantly up-regulated in the SHR+timisartan,SHR+CAD,and timisartan+CAD groups,compared with the SHR+NC group(P<0.05).The results of qRT-PCR showed that the expression level of miR-19b-3p in aorta of SHR rats was significantly decreased in VSMC cells in the temesartan+NC,NC+CAD and temesartan+CAD groups compared with the control group(P<0.05).The results of transwell assay showed that the migration of VSMC cells was significantly decreased in the temosartan+NC,NC+CAD and temosartan+CAD groups compared with that in the control group(P<0.05).Compared with the blank group,the expression levels of IL-1βand TNF-αwere significantly reduced in the miR-19b-3p inhibitor group,which was enhanced by further addition of temosartan and calcium hydroxybenzenesulfonate treatment(P<0.05).Conclusion Telmisartan combined with Calcium Dobesylate to relieve the aortic-injury and inhibitor the expression of inflammatory factors in spontaneous hypertension via the miR-19b-3p/SOCS1 axis.
作者
田春辉
胡威威
娄满
胡晓英
田心怡
高春燕
TIAN Chunhui;HU Weiwei;LOU Man;HU Xiaoying;TIAN Xinyi;GAO Chunyan(Department of Geriatrics,Hengshui People's Hospital,Hengshui City,Hengshui 053000,Hebei,China;Department of Endocrinology,Affiliated Hospital of Qingdao University,Qingdao 266400,Shandong,China)
出处
《医学研究与战创伤救治》
CAS
北大核心
2024年第3期254-260,共7页
Journal of Medical Research & Combat Trauma Care
基金
衡水市科学技术研究与发展计划项目(2021014089Z)。
