新疆鼠尾草中6,7-去氢罗列酮对凝血酶诱导人脐静脉内皮细胞损伤的保护作用及机制  

Protective effects and mechanism of 6,7-Dehydroroyleanone from Salvia deserta Schang on HUVEC injuried by thrombin

作　　者：刘阳 帕提曼·肖开提 穆巴热科·吾普尔 王晓梅[1] 胡君萍[1] 王新玲[1] LIU Yang;Patiman Xiaokaiti;Mubareke Wupuer;WANG Xiaomei;HU Junping;WANG Xinling(Department of Natural Medicine Chemistry Teaching and Reseurch College of Pharmacy of Xinjiang Medical University, Urumqi 830000, China)

机构地区：[1]新疆医科大学药学院天然药物化学教研室,乌鲁木齐830011

出　　处：《新疆医科大学学报》2020年第7期890-894,900,共6页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2016D01C160)。

摘　　要：目的通过体外实验研究新疆鼠尾草中6,7-去氢罗列酮对凝血酶诱导损伤的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)的保护作用及可能的机制。方法对HUVEC进行体外培养,通过凝血酶诱导HUVEC建立损伤模型,分为空白组、模型组(30 U/mL凝血酶)、6,7-去氢罗列酮组(0.1、0.2、0.3 mg/mL)预孵育24 h、肝素组(1 mg/mL)预孵育24 h,分别采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)法检测6,7-去氢罗列酮对30 U/mL凝血酶损伤4 h的细胞活力;倒置显微镜下观察细胞形态;酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测细胞裂解液中血管性血友病因子(von willebrand factor,vWF)、纤溶酶原激活物抑制物-1(plasminogen activator inhibitor-1,PAI-1)和组织型纤溶酶原激活物(Tissue plasminogen activator,t-PA)的含量以及t-PA/PAI-1的比值;实时荧光定量法(real-time quantitative PCR,QRT-PCR)检测vWF、PAI-1、t-PA的分泌及其mRNA的表达,并比较4组异同。结果与空白组比较,凝血酶损伤后的HUVEC活力明显受到抑制(P<0.05),细胞呈不规则形态;与模型组比较,肝素组和6,7-去氢罗列酮组明显提高细胞损伤后的活力,凝血酸损伤的HUVEC形态逐渐恢复至正常的纺锤形态,并能显著上调t-PA的含量及t-PA/PAI-1的比值(P<0.05)。6,7-去氢罗列酮高、中剂量组能够明显下调PAI-1和vWF的含量及其表达。结论6,7-去氢罗列酮对凝血酶诱导损伤的HUVEC具有保护作用,其机制可能与提高细胞纤溶活性,维持血管内皮细胞的正常功能有关。Objective The protective effect of 6,7-dehydroroyleanone from Salvia deserta Schangon thrombin-induced injury of the human umbilical vein endothelial cells(HUVEC)and its possible mechanism were studied in vitro.Methods HUVEC was cultured in vitroand its injured model was induced by thrombin.This study was divided into as follows:normal group,30 U/mL thrombin injury model group,6,7-dehydroroyleanone protective groups(0.1,0.2,0.3 mg/L)incubation for 24 hours and heparin group(1 mg/mL)incubation for 24 hours.MTT method was used to detect cell activity that was protected by 6,7-dehydroroyleanone from damage caused by 30 U/mL thrombin for 4 hours.Cell morphology was observed under inverted microscope.The levels of von Willebrand factor(vWF),plasminogen activator inhibitor-1(PAI-1)and tissue plasminogen activator(t-PA)were detected by ELISA.The secretion of vWF,PAI-1 and t-PA and their expression were detected by real-time fluorescence quantitative method(PCR).Results Compared with the blank group,HUVEC activity was significantly inhibited after thrombin injury(P<0.05),and the cells were showed irregularmorphology.The activityof HUVEC in heparin group and 6,7-dehydroroyleanone group was significantly increased after thrombin injury,and its cell morphology was gradually returned to normal spindle morphology.Compared with themodel group,6,7-dehydroroyleanone could significantly increase the content of t-PA and its expression and t-PA/PAI-1 ratio(P<0.05),the medium and high dose groups of 6,7-dehydroroyleanone can significantly reduce the contents and their expression of t-PA and vWF.Conclusion 6,7-dehydroroyleanone has protective effect on thrombin-induced HUVEC injury,and its mechanism may be related to improving the fibrinolytic activity of cells and maintaining the normal function of vascular endothelial cells.

关 键 词：新疆鼠尾草 6 7-去氢罗列酮 凝血酶 人脐静脉内皮细胞 

分 类 号：R285.5[医药卫生—中药学]