结蛋白介导的雌激素受体对颅内动脉瘤血管平滑肌细胞表型的调控作用  

Effect of desmin-mediated estrogen receptors on phenotype regulation of vascular smooth muscle cells in intracranial aneurysms

作　　者：隋昕 隋天意 张旭[3] 冯旭 王东玉[4] SUI Xin;SUI Tianyi;ZHANG Xu;FENG Xu;WANG Dongyu(Department of Neurosurgery,The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China;People’s Liberation Army General Hospital Graduate School,Beijing 100000,China;Jinzhou Center for Disease Control and Prevention,Jinzhou 121000,China;Department of Neurology,Jinzhou Central Hospital,Jinzhou 121000,China)

机构地区：[1]锦州医科大学附属第一医院神经外科,辽宁锦州121000 [2]中国人民解放军总医院研究生院,北京100000 [3]锦州市疾病预防控制中心,辽宁锦州121000 [4]锦州市中心医院神经内科,辽宁锦州121000

出　　处：《中国医科大学学报》2024年第12期1094-1098,1106,共6页Journal of China Medical University

基　　金：辽宁省教育厅科学研究经费项目(KJKZ0812)。

摘　　要：目的探讨颅内动脉瘤组织中结蛋白的表达及其通过雌激素受体(ER)对血管平滑肌细胞(VSMC)表型调控的影响。方法选取非动脉瘤患者手术区颈外动脉分支(正常动脉血管)和颅内动脉瘤切除标本28例。采用免疫组织化学染色法检测正常动脉血管和颅内动脉瘤中结蛋白的表达情况,采用Western blotting和实时定量PCR检测颅内动脉瘤组织及正常血管中结蛋白和DES mRNA的表达。将VSMC分为control组、shNC组、shDesmin组、pcDNA3.1/NC组、pcDNA3.1/Desmin组和pcDNA3.1/Desmin+3-MA组。shNC组、shDesmin组、pcDNA3.1/NC组、pcDNA3.1/Desmin组分别转染shNC、shDesmin、pcDNA3.1/NC和pcDNA3.1/Desmin质粒,pcDNA3.1/Desmin+3-MA组在转染前加入10μmol/L 3-MA预处理6 h。采用实时定量PCR和Western blotting检测VSMC中DES mRNA和结蛋白的表达;用CCK-8实验检测沉默DES对VSMC增殖能力的影响;用Transwell实验检测沉默DES对VSMC迁移能力的影响;用Western blotting检测沉默DES对VSMC中ERα/ERβ比例和表型调节相关蛋白表达水平的影响。结果结蛋白在颅内动脉瘤组织中的阳性表达率低于正常动脉血管(χ^(2)=16.601,P<0.001);与shNC组比较,shDesmin组VSMC中结蛋白表达水平降低(P<0.001),细胞增殖力增强(P<0.05),细胞迁移数量增多(P<0.001),ERα/ERβ比例升高(P<0.001);与pcDNA3.1/NC组比较,pcDNA3.1/Desmin组VSMC中人平滑肌α肌动蛋白(SM-α-actin)、人平滑肌肌球蛋白重链(SM-MHC)蛋白表达水平升高(P<0.01),基质金属蛋白酶-3(MMP-3)、肿瘤坏死因子-α(TNF-α)蛋白表达水平下降(P<0.05)。与pcDNA3.1/Desmin组比较,pcDNA3.1/Desmin+3-MA组VSMC中SM-α-actin、SM-MHC蛋白表达水平下降(P<0.05),MMP-3、TNF-α蛋白表达水平升高(P<0.05)。结论结蛋白在颅内动脉瘤组织中低表达。沉默DES促进VSMC增殖、迁移,进而可能促进VSMC表型调节。Objective To investigate desmin expression in intracranial aneurysm tissue and its mediating effect on the phenotype regulation of vascular smooth muscle cell(VSMC)by estrogen receptor(ER).Methods Twenty-eight samples of surgical site specimens including external carotid artery branches and intracranial aneurysm resection were collected from patients with nonarterial aneurysms in clinical practice.Desmin expression in the arterial blood vessels and intracranial aneurysms was detected using immunohistochemical staining(SABC),whereas desmin expression in the intracranial aneurysm tissue and normal blood vessels was detected using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blotting.The VSMC were divided into control,shNC,shDesmin,pcDNA3.1/NC,pcDNA3.1/Desmin,and pcDNA3.1/Desmin+3-MA groups.The shNC,shDesmin,pcDNA3.1/NC,and pcDNA3.1/Desmin groups were transfected with shNC,shDesmin,pcDNA3.1/NC,or pcDNA3.1/Desmin plasmids,respectively.The pcDNA3.1/Desmin+3-MA group was pretreated with 10μmol/L 3-MA for 6 hours before transfection was performed.qRT-PCR and Western blotting were used to detect the expression of silenced DES in VSMC.CCK-8 assay was used to detect the effect of silencing DES on the proliferation ability of VSMC.Transwell cell migration assay was used to detect the effect of silencing DES on the migration ability of VSMC.Western blotting was used to detect the effects of DES silencing on the ERα/ERβratio,and phenotype regulatory protein levels in VSMC.Results Immunohistochemical staining showed that the positive expression rate of desmin in intracranial aneurysms was significantly lower than that in normal arterial blood vessels(χ^(2)=16.601,P<0.001).Compared with normal vascular tissue,desmin expression in intracranial aneurysm tissue was reduced(P<0.001).Compared with the shNC group,the expression level of desmin in VSMC of the shDesmin group decreased(P<0.001),the cell proliferation was stronger(P<0.05),the number of cell migrations increased(P<0.001),and the ER

关 键 词：颅内动脉瘤 结蛋白 血管平滑肌细胞 雌激素 表型调节 

分 类 号：R651[医药卫生—外科学]