香青兰有效部位对OGD/R损伤诱导的人脑微血管内皮细胞保护作用研究  

Study on the Protective Effect of Effective Parts of Dracocephalum moldavica on Human Microvascular Endothelial Cells after OGD/R Injury

作　　者：杨志航 卡德尔业·卡德尔 徐磊 徐明[4] 郑瑞芳 邢建国 YANG Zhi-hang;KADDER Kadeerye;XU Lei;XU Ming;ZHENG Rui-fang;XING Jian-guo(College of Pharmacy,Shihezi University,Shihezi 832000,China;Xinjiang Institute of Materia Medica,Urumqi 830002,China;Key Laboratory of Xinjiang Uyghur Medicine Urumqi 830002,China;China Pharmaceutical University,Nanjing 210009,China)

机构地区：[1]石河子大学药学院,新疆石河子832000 [2]新疆药物研究所,乌鲁木齐830002 [3]新疆维吾尔药重点实验室,乌鲁木齐830002 [4]中国药科大学,南京210009

出　　处：《中国药学杂志》2022年第11期918-924,共7页Chinese Pharmaceutical Journal

基　　金：新疆维吾尔自治区青年科技人才-乡村振兴项目资助(WJWV-XCzX202205);中央引导地方科技发展专项资助(ZYYD2022A02);新疆维吾尔自治区重点实验室开放课题资助(2020D04020);新疆维吾尔自治区公益性科研院所科研业务费资助(KY2018137);2019年新疆维吾尔自治区高层次人才引进工程(柔性引进人才)项目资助。

摘　　要：目的 采用泛半胱氨酸天冬氨酸蛋白酶(cysteine aspartic acid protease, Caspase)抑制剂Z-VAD-FMK联合氧糖剥夺/复氧(oxygen-glucose deprivation/re-oxygenation, OGD/R)建立体外脑缺血再灌注损伤模型,探讨了香青兰有效部位(effective parts of Dracocephalum moldavica, EPDM)对人脑微血管内皮细胞(human brain microvascular endothelial cells, HBMECs)的保护作用。方法 CCK-8法检测细胞活力,倒置显微镜下观察细胞形态,NO测定试剂盒检测NO含量,transwell法检测细胞迁移情况,免疫荧光检测血管内皮-钙黏蛋白(VE-Cadherin)的表达情况,免疫印迹检测酪氨酸激酶(SRC)、内皮素-1(ET-1)、紧密连接蛋白Claudin 5、内皮性一氧化氮合酶(eNOS)、及磷酸化eNOS的蛋白表达。结果 与对照组比较,Z-VAD-FMK联合OGD/R可使HBMECs活力下降,细胞形态变差,细胞迁移减少,NO释放量降低,VE-cadherin表达减少,Src、Claudin 5、磷酸化eNOS表达减少,ET-1表达升高;而EPDM预处理可部分逆转Z-VAD-FMK联合OGD/R引起的上述因素的变化。结论 EPDM对体外脑缺血再灌注模型下的HBMECs具有保护作用,可促进细胞增殖、迁移,增强血管舒缩调节能力,并通过增强紧密连接和黏附连接维持屏障功能。OBJECTIVE To investigate the protective effect of EPDM on endothelial cells, a cerebral ischemia-reperfusion injury model established by treatment with the pan-cysteine aspartic acid protease(Caspase) inhibitor Z-VAD-FMK combined with oxygen-glucose deprivation/re-oxygenation(OGD/R). METHODS Cell Counting Kit-8(CCK-8) was used to detect cell viability, cell morphology was observed by microscope, NO content was detected by NO determination kit, and cell migration was detected by Transwell method. The expression of vascular endothelium-cadherin was detected by immunofluorescence. Western blot was used to detect the protein expressions of non-receptor tyrosine kinase(Src), endothelin-1(ET-1), blood-brain barrier tight junction associated protein 5(Claudin 5), endothelial nitric oxide synthase(eNOS), and phosphorylated eNOS. RESULTS Compared with the control group, Z-VAD-FMK combined with OGD/R reduced HBMECs activity, cell morphology, cell migration, NO release, VE-cadherin expression, SRC, Claudin 5 and p-eNOS expression, andincrease ET-1 expression. EPDM pretreatment can partially reverse the above factors induced by Z-VAD-FMK combined with OGD/R. CONCLUSION EPDM has a protective effect on HBMECs under cerebral ischemia reperfusion model, which can promote cell proliferation and migration, enhance vasomotor regulation, and maintain barrier function by enhancing tight junctions and adhesion junctions.

关 键 词：香青兰有效部位 人脑微血管内皮细胞 脑缺血再灌注损伤 氧糖剥夺/复氧 屏障功能 

分 类 号：R965[医药卫生—药理学]