雌激素对同型半胱氨酸损伤小鼠脑微血管内皮细胞的保护作用  

Protective effect of estrogen on mouse cerebral microvascular endothelial cells after homocysteine injury

作　　者：余雅欢 高稳 王娟 Yu Yahuan;Gao Wen;Wang Juan(School of Public Health,Guangzhou Medical University,Guangzhou 511436,Guangdong Province,China)

机构地区：[1]广州医科大学公共卫生学院,511436

出　　处：《中华老年心脑血管病杂志》2024年第6期704-708,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81703299);广东省自然科学基金(2022A1515012167)。

摘　　要：目的探讨雌激素对同型半胱氨酸(Hcy)损伤小鼠脑微血管内皮细胞株(bEnd.3)后基质金属蛋白酶(MMP)-9、MMP-2和紧密连接蛋白Occludin表达的影响及其对脑小血管病的保护作用和机制。方法采用Hcy、β-雌二醇分别处理bEnd.3细胞,以细胞增殖及毒性检测试剂盒确定最适药物浓度。实验细胞分为对照组(仅加培养液)、Hcy组(200μmol/L的Hcy)、β-雌二醇组(200 nmol/L的β-雌二醇)、联合组(200 nmol/L的β-雌二醇处理30 min后再加入200μmol/L的Hcy),各组均处理6 h。Western blot法检测各组细胞中MMP-9、MMP-2和Occludin表达。结果各组24 h、48 h的MMP-9、MMP-2和Occludin表达比较,差异无统计学意义(P>0.05)。与对照组比较,Hcy组72 h的MMP-9和MMP-2表达明显升高(1.45±0.14 vs 1.00±0.00,1.35±0.15 vs 1.00±0.00,P<0.05),Occludin表达明显降低(0.64±0.05 vs 1.00±0.00,P<0.05)。与Hcy组比较,联合组72 h的MMP-9和MMP-2表达明显降低(0.84±0.19 vs 1.45±0.14,1.01±0.78 vs 1.35±0.15,P<0.05),Occludin表达明显升高(0.91±0.10 vs 0.64±0.05,P<0.05)。结论在Hcy处理的bEnd.3中β-雌二醇能够改善血脑屏障损伤,从而导致MMP-9和MMP-2表达降低,Occludin表达升高。Objective To investigate the effect of estrogen on the expression of matrix metalloproteinase protein 9 and 2(MMP-9,MMP-2)and tight junction protein Occludin in mouse brain microvascular endothelium cell line bEnd.3 injured by homocysteine(Hcy),and explore the protective effects and underlying mechanisms of estrogen on small cerebral vascular diseases.Methods bEnd.3 cells were treated with Hcy andβ-estradiol respectively,and the optimal concentration was determined for cell proliferation and toxicity with enhanced cell counting kit-8.The cells were divided into control group,Hcy group(200μmol/L),E2βgroup(200 nmol/L)and combined treatment group(200 nmol/L E2βtreatment for 30 min followed by 200μmol/L Hcy).After 6 h corresponding treatment,Western blotting was used to detect the protein levels of MMP-9,MMP-2 and Occludin in each group of cells.Results There were no significant differences in the protein levels of MMP-9,MMP-2 and Occludin in above groups after treatment for 24 and 48 h(P>0.05).Compared with the control group,the expression levels of MMP-9 and MMP-2 was significantly increased in the Hcy Group after treatment of 72 h(1.45±0.14 vs 1.00±0.00,1.35±0.15 vs 1.00±0.00,P<0.05),while the level of Occludin was obviously decreased(0.64±0.05 vs 1.00±0.00,P<0.05).Compared in the Hcy Group,the MMP-9 and MMP-2 levels were obviously decreased(0.84±0.19 vs 1.45±0.14,1.01±0.78 vs 1.35±0.15,P<0.05),and that of Occludin was notably elevated in the combined treatment group(0.91±0.10 vs 0.64±0.05,P<0.05).Conclusion E2βcan ameliorate blood-brain barrier damage in Hcy-treated bEnd.3 cells,resulting in decreased MMP-9 and MMP-2 expressions and increased Occludin expression.

关 键 词：雌二醇 高半胱氨酸 大脑小血管疾病 血管内皮细胞 基质金属蛋白酶9 紧密连接蛋白质类 

分 类 号：R743[医药卫生—神经病学与精神病学]