人参皂苷Rb1调节脑微血管NOS/NO改善脑细胞膜通透性  

Ginsenoside Rb1 Regulates Brain Microvascular NOS/NO Improves Brain Cell Membrane Permeability

作　　者：马琪 张小玲[1] 王春亚[1] 郭蕾[1] 赵玉杰[1] 侯彦丽[1] 骆燕妮 王小闯[1] MA Qi;ZHANG Xiao-ling;WANG Chun-ya;GUO Lei;ZHAO Yu-jie;HOU Yan-li;LUO Yan-ni;WANG Xiao-chuang(Department of Critical Care Medicine, the Second Affiliated Hospital of Xi'an Jiangtong University Medical College,Xi'an, Shaanxi, 710014, China)

机构地区：[1]西安交通大学第二附属医院重症医学科,陕西西安710014

出　　处：《现代生物医学进展》2019年第8期1430-1435,共6页Progress in Modern Biomedicine

基　　金：西安交通大学第二附属医院科研基金项目(YJ(QN)201709);国家自然科学基金项目(81670049)

摘　　要：目的:探究人参皂苷Rb1在脂多糖(LPS)诱导的人脑微血管细胞模型中对脑细胞膜通透性的影响。方法:采用噻唑蓝比色法(MTT)及乳酸脱氢酶(LDH)释放检测不同浓度人参皂苷Rb1对LPS刺激人脑微血管内皮细胞系(HBMEC)存活率的影响;采用EVOM法检测HBMEC细胞通透性;采用Griess法、ELISA法及DAF-FM DA荧光探针分别检测NO、ONOO^-含量及eNOS、iNOS活性;Western blot法检测p-eNOS(ser1177)、iNOS蛋白的表达水平。结果:与正常对照组比较,LPS(5μg/m L)可明显降低HBMEC细胞存活率(P<0.01);与LPS组比较,随着人参皂苷Rb1浓度的增加细胞存活率明显升高,且中、高剂量组(40、80μmol/L)具有显著性差异(P<0.05、P<0.01);LPS导致HBMEC单层细胞电阻值明显降低(P<0.01),人参皂苷Rb1高剂量组(80μmol/L)可显著抑制LPS所致的细胞膜通透性升高(P<0.05);LPS可明显降低HBMEC细胞NO含量(P<0.01)升高ONOO^-含量(P<0.05),人参皂苷Rb1中、高剂量组(40、80μmol/L)较LPS组具有显著升高NO含量,降低ONOO^-含量的作用(P<0.05);与正常对照组比较,LPS组可明显降低HBMEC细胞磷酸化eNOS (ser1177)蛋白的表达水平,而显著升高iNOS蛋白表达水平(P<0.01)。人参皂苷Rb1高剂量组(80μmol/L)较LPS组可显著升高p-eNOS(ser1177)蛋白的表达水平,降低iNOS蛋白的表达水平(P<0.05)。结论:人参皂苷Rb1可有效降低iNOS活性,升高eNOS活性继而减少NO水平及ONOO^-含量,显著降低LPS导致的脑细胞膜通透性升高。Objective: To investigate the effect of ginsenoside Rb1 on brain cell membrane permeability in lipopolysaccharide (LPS) induced brain microvascular cell model. Methods: Methylthiazolyldiphenyl tetrazolium bromide(MTT) and lactate dehydrogenase (LDH) release assays were used to detect the survival rate of different concentrations of ginsenoside Rb1 on of human brain microvascular endothelial cell line(HBMEC) stimulated by LPS;The HBMEC cell permeability by epithelial voltmeter(EVOM)method;The content of nitric oxide(NO), peroxynitrite(ONOO^-) and the activity of eNOS, i NOS were detect respectively by Griess method, ELSA and fluorescence spectrophotometer DAF-FM DA. The expression of protein levels of phosphorylated endothelial nitricoxide synthase(p-eNOS Ser1177) and inducible nitric oxide synthase(iNOS) were detected by Western blot. Results: Compared with the control group, LPS(5 μg/m L) could significantly reduce the survival rate of HBMEC cells(P<0.01), Compared with the LPS group, the cell survival rate increased significantly with the ginsenoside Rb1 concentration increased, and the medium and high dose groups(40, 80 μmol/L) had significant difference(P<0.05, P<0.01);LPS caused the transendothelial electrical resistance of HBMEC cells was significantly decreased(P <0.01), and ginsenoside Rb1 high dose group(80 μmol/L) could significantly decrease the cell membrane permeability induced by LPS(P<0.05). LPS could significantly reduce the content of NO(P<0.01) and increase the content of ONOO^-(P<0.05) in HBMEC cells, the ginsenoside Rb1 medium and high dose group(40, 80 μmol/L) had significantly increased NO content and decreased ONOO^-content compared with LPS group(P<0.05). Compared with the control group, LPS group significantly decreased the expression of phosphorylated eNOS(ser1177) protein in HBMEC cells, and significantly increased the level of i NOS protein expression(P<0.01). Ginsenoside Rb1 high dose group(80 μmol/L) can significantly increase the expression level of p-eNOS(ser1177) protein and

关 键 词：人参皂苷RB1 人脑微血管内皮细胞 细胞膜通透性 NOS/NO 

分 类 号：R-33[医药卫生] Q593.2[生物学—生物化学]