川芎嗪对帕金森病炎症细胞模型增殖和凋亡的影响及机制  

Effect and mechanism of ligustrazine on proliferation and apoptosis of inflammatory cell models in Parkinson's disease

作　　者：王霆[1] 吴穹[2] 季帅 WANG Ting;WU Qiong;JI Shuai(Department of Neurology,Jiyuan People's Hospital,Jiyuan 459000,Henan Province,China;Function Laboratory,Department of Medicine of Qinghai University,Xining 810001,Qinghai Province,China)

机构地区：[1]济源市人民医院神经内科,河南济源459000 [2]青海大学医学部机能实验室,青海西宁810001

出　　处：《新乡医学院学报》2024年第12期1114-1124,共11页Journal of Xinxiang Medical University

基　　金：第三次中国国家卒中登记研究及亚组课题研究(编号:D151100002015003)。

摘　　要：目的探讨川芎嗪对帕金森病炎症细胞模型增殖和凋亡的影响及机制。方法将对数生长期人小胶质细胞HMC3随机分为对照组、脂多糖(LPS)组、LPS+12.5μmol·L^(-1)川芎嗪组、LPS+25.0μmol·L^(-1)川芎嗪组、LPS+50.0μmol·L^(-1)川芎嗪组、LPS+100.0μmol·L^(-1)川芎嗪组。对照组细胞不给予任何药物进行干预;LPS组、LPS+12.5μmol·L^(-1)川芎嗪组、LPS+25.0μmol·L^(-1)川芎嗪组、LPS+50.0μmol·L^(-1)川芎嗪组、LPS+100μmol·L^(-1)川芎嗪组细胞首先给予1 mg·L^(-1) LPS刺激24 h;LPS+12.5μmol·L^(-1)川芎嗪组、LPS+25.0μmol·L^(-1)川芎嗪组、LPS+50.0μmol·L^(-1)川芎嗪组、LPS+100.0μmol·L^(-1)川芎嗪组细胞分别加入12.5、25.0、50.0、100.0μmol·L^(-1)川芎嗪干预24 h;采用活细胞计数试剂盒-8法检测各组细胞活力,筛选出保持细胞活力最佳的川芎嗪干预浓度。将对数生长期HMC3细胞随机分为对照组、LPS组、LPS+mimic NC组、LPS+miR-145 mimic组、LPS+inhibitor NC组、LPS+miR-145 inhibitor组、川芎嗪+LPS组、川芎嗪+LPS+miR-145 mimic组及川芎嗪+LPS+miR-145 inhibitor组;对照组细胞不给予任何药物进行干预,LPS组细胞给予1 mg·L^(-1) LPS刺激24 h,LPS+mimic NC组、LPS+miR-145 mimic组、LPS+inhibitor NC组及LPS+miR-145 inhibitor组细胞应用脂质体2000转染试剂分别转染mimic NC、miR-145 mimic、inhibitor NC、miR-145 inhibitor并给予1 mg·L^(-1) LPS刺激24 h,川芎嗪+LPS组细胞给予1 mg·L^(-1) LPS刺激24 h并给予100.0μmol·L^(-1)川芎嗪干预,川芎嗪+LPS+miR-145 mimic组、川芎嗪+LPS+miR-145 inhibitor组细胞首先应用脂质体2000转染试剂分别转染miR-145 mimic、miR-145 inhibitor并给予1 mg·L^(-1) LPS、100.0μmol·L^(-1)川芎嗪干预24 h;采用实时荧光定量聚合酶链式反应实验检测细胞中miR-145相对表达量,5-乙炔基-2′-脱氧尿苷实验检测细胞增殖情况,Hoechst 33258染色试验检测细胞凋亡情况,酶联免疫吸附实验检测各组细胞上清液中肿瘤�Objective To explore the effect and mechanism of ligustrazine on proliferation and apoptosis of inflammatory cell models in Parkinson's disease.Methods Human microglial cells(HMC3)in logarithmic growth phase were randomly divided into control group,lipopolysaccharide(LPS)group,LPS+12.5μmol·L^(-1) ligustrazine group,LPS+25.0μmol·L^(-1) ligustrazine group,LPS+50.0μmol·L^(-1) ligustrazine group,and LPS+100.0μmol·L^(-1) ligustrazine group.The cells in the control group did not receive any drug intervention.The cells in the LPS group,LPS+12.5μmol·L^(-1) ligustrazine group,LPS+25.0μmol·L^(-1) ligustrazine group,LPS+50.0μmol·L^(-1) ligustrazine group,and LPS+100.0μmol·L^(-1) ligustrazine group were first stimulated with 1 mg·L^(-1) LPS for 24 hours;the cells in the LPS+12.5μmol·L^(-1) ligustrazine group,LPS+25.0μmol·L^(-1) ligustrazine group,LPS+50.0μmol·L^(-1) ligustrazine group,and LPS+100.0μmol·L^(-1) ligustrazine group were treated with 12.5,25.0,50.0,and 100.0μmol·L^(-1) ligustrazine for 24 hours,respectively.The viability of cells in each group was detected by using the live cell counting kit-8,and the optimal concentration of ligustrazine was selected to intervene in cell viability.The HMC3 cells in logarithmic growth phase were randomly divided into control group,LPS group,LPS+mimic NC group,LPS+miR-145 mimic group,LPS+inhibitor NC group,LPS+miR-145 inhibitor group,ligustrazine+LPS group,ligustrazine+LPS+miR-145 mimic group,and ligustrazine+LPS+miR-145 inhibitor group.The cells in the control group were not intervened with any drugs;the cells in the LPS group were stimulated with 1 mg·L^(-1) LPS for 24 hours;the cells in the LPS+mimic NC group,LPS+miR-145 mimic group,LPS+inhibitor NC group,and LPS+miR-145 inhibitor group were transfected with mimic NC,miR-145 mimic,inhibitor NC,and miR-145 inhibitor by liposomal 2000 reagent,respectively,and then were stimulated with 1 mg·L^(-1) LPS for 24 hours;the cells in the ligustrazine+LPS group were first stimulated with 1 mg·L^(-1) LPS for 2

关 键 词：帕金森病 川芎嗪 微RNA-145 炎症细胞 增殖 凋亡 磷脂酰肌醇3-激酶 蛋白激酶B 

分 类 号：R742.5[医药卫生—神经病学与精神病学]