摘要
目的在细胞水平探索补肾活血方干预高糖高脂诱导的小鼠脑微血管内皮细胞损伤的研究。方法在体外实验中采用高糖高脂刺激小鼠脑微血管内皮细胞。实验分组为:空白对照组、模型组[25 mmol/L葡萄糖(HG),200μmol/L棕榈酸(PA)]、补肾活血方30、60、120 mg/L组。CCK-8细胞增殖实验方法检测补肾活血方对高糖高脂诱导的细胞存活率的影响;采用试剂盒检测各组细胞部分氧化应激因子[活性氧(ROS)、诱导型一氧化氮合酶(iNOS)、一氧化氮(NO)、血管内皮生长因子(VEGF)]、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶抑制剂1(TIMP-1)和部分炎症因子[IL-6、细胞黏附因子1(ICAM-1)、TNF-α、转化生长因子β1(TGF-β1)]的含量变化。采用实时荧光定量PCR法检测iNOS和TNF-αmRNA表达。结果(1)与空白对照组(100%±2.3%)比较,模型组细胞的细胞存活率显著降低(62.6%±0.5%,P<0.01);与模型组比较,补肾活血方30、60、120 mg/L组细胞的存活率显著升高[(70.5%±4.0%),(76.3%±2.8%),(80.5%±2.7%),P<0.05,P<0.01]。(2)与空白对照组比较,模型组细胞的部分氧化应激因子(ROS、iNOS、NO、VEGF)、MMP-2、MMP-9、TIMP-1以及炎症相关因子(IL-6、ICAM-1、TNF-α、TGF-β1)含量显著上升(P<0.05,P<0.01)。与模型组比较,补肾活血方30、60、120 mg/L组作用后各检测因子的表达降低(P<0.05,P<0.01)。(3)荧光定量PCR结果显示,与空白对照组比较,模型组iNOS和TNF-αmRNA表达均增加(P<0.01),与模型组比较,BSHX 120 mg/L组iNOS mRNA表达及BSHX 60、120 mg/L组TNF-αmRNA表达均降低(P<0.05,P<0.01)。结论高糖高脂能够促进ROS、iNOS、NO、VEGF、MMP-2、MMP-9、TIMP-1和部分炎症因子等相关指标的表达,使细胞活力降低,而补肾活血方可以改善高糖高脂诱导的小鼠脑微血管内皮细胞的损伤。
Objective To explore the effect of Bushen Huoxue(BSHX)prescription on prevention and treatment of mouse brain microvascular endothelial cells injury induced by high glucose and hyperlipemia at the cellular level.Methods In vitro experiments,high glucose(HG,25 mmol/L)and palmitic acid(PA,200μmol/L)were used to intervene in the mouse brain microvascular endothelial cells to establish the cell damage model.The cells were divided into control group,model group[25 mmol/L high glucose(HG)+200μmol/L palmitic acid(PA)]and BSHX(30,60,120 mg/L)groups.The effect of BSHX on hyperglycemic and hyperlipid-induced cell damage was detected by cell counting kit-(CCK-)8 assay.The changes of oxidative stress related factors[reactive oxygen species(ROS),inducible nitric oxide synthase(iNOS),vascular endothelial growth factor(VEGF)],matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),tissue inhibitor of metalloproteinases-1(TIMP-1)]and inflammatory factors[IL-6,intercellular adhesion molecule 1(ICAM-1),TNF-α,transforming growth factorβ1(TGF-β1)]were detected by ELISA.The cytokine NO was determined by biochemical kit assay.The expressions of iNOS and TNF-αmRNA were detected by quantitative real-time PCR.Results(1)After treatment,compared with the control group(100%±2.3%),the survival rate of the cells in the model group was significantly reduced(62.6%±0.5%,P<0.01).Compared with the model group,the survival rate of the cells in the BSHX 30,60,120 mg/L groups were significantly increased[(70.5%±4.0%),(76.3%±2.8%),(80.5%±2.7%),P<0.05,P<0.01].(2)Compared with the control group,the expression of oxidative stress and inflammatory related factors(ROS,iNOS,NO,VEGF,MMP-2,MMP-9,TIMP-1,IL-6,ICAM-1,TNF-α,TGF-β1)in the model group increased significantly(P<0.05,P<0.01).Compared with the model group,the expression of each detection factor was decreased in the BSHX 30,60,120 mg/L groups.(3)Compared with the control group,the expressions of iNOS and TNF-αmRNA in the model group were increased(P<0.01),and the expression of
作者
赵佳奇
张益铭
徐飞飞
王梓林
李晓青
郭美彤
王洁
李钦青
田雅娟
贺文彬
ZHAO Jia-qi;ZHANG Yi-ming;XU Fei-fei;WANG Zi-ling;LI Xiao-qing;GUO Mei-tong;WANG Jie;LI Qin-qing;TIAN Ya-juan;HE Wen-bin(Shanxi Province Key Laboratory of Chinese Medicine Encephalop-athy,Shanxi University of Chinese Medicine,Taiyuan,030619)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2020年第8期941-947,共7页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81530099,No.81473375)
山西省留学回国人员科研项目(No.2013-134)。
关键词
补肾活血
高糖高脂
氧化应激
炎症
Bushen Huoxue Prescription
high glucose and high palmitate
oxidative stress
inflammatory
