黄苓昔对脂多糖诱导的H9C2心肌细胞炎症的保护作用及机制研究  

Protective effect of Baicalin on lipopolysaccharide induced H9C2 cell inflammation and its mechanism

作　　者：李萌芳[1] 陈隆望[1] 胡系意 郑来赞 连洁[1] 赵光举[1] 邱俏檬[1] 洪广亮[1] 卢中秋[1] Li Mengfang;Chen Longwang;Hu Xiyi;Zhen Laizan;Lian Jie;Zhao Guangju;Qiu Qiaomeng;Hong Guangliang;Lu Zhongqiu(Emergency Department, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China)

机构地区：[1]温州医科大学附属第一医院急诊医学中心,325000

出　　处：《中华急诊医学杂志》2019年第8期983-988,共6页Chinese Journal of Emergency Medicine

基　　金：浙江省自然科学基金(LQ15H150003;LY15H150006);国家自然科学基金(81772112;81571937;81871583);浙江省医药卫生科研项目(2015KYA152);浙江省中医药管理局项目(2015ZA120);温州市科技计划项目(Y20170177;Y20170185;Y20150182).

摘　　要：目的探讨黄芩苷对脂多糖(Lipopolysaccharides,LPS)诱导的H9C2心肌细胞炎症反应的保护作用及可能机制。方法培养H9C2心肌细胞,用终浓度为10、20、30 μmol/L黄芩苷预处理12 h后,加入终浓度为1 μg/mL的LPS刺激6 h,对照组加入等量的生理盐水,收集细胞样本。CCK-8(The Cell Counting Kit-8)检测细胞活力,酶联免疫吸附法(ELISA)检测白介素-6(IL-6)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的表达水平,蛋白免疫印迹法(Western blot)检测细胞中NF-κB p65、磷酸化NF-κB p65(p-NF-κB p65)、p38 MAPK、磷酸化p38 MAPK(p-p38 MAPK)、IκBα、磷酸化IκBα(p-IκBα)的蛋白表达水平的变化。采用SPSS 23.0统计软件,两组样本比较采用独立样本t检验,多组间比较采用单因素方差分析检验。结果对照组心肌细胞存活率为(93.67±1.453)%,而LPS刺激组存活率为(60.33±2.603)%。与对照组相比,LPS诱导后H9C2心肌细胞存活率降低(P<0.05)。对照组H9C2心肌细胞IL-6表达量为(49.33±2.42)pg/mL,TNF-a表达量为(86.33±1.85)pg/mL,IL-1β表达量为(28.67±4.66)pg/ml。与对照组相比,LPS诱导后H9C2心肌细胞IL-6、IL-1β及TNF-α表达水平升高(P<0.05),p-NF-κB p65、p-p38 MAPK、p-IκBα蛋白水平升高(P<0.05),IκBα蛋白水平降低(P<0.05),而NF-κB p65及p38 MAPK蛋白表达无明显变化(P>0.05)。与LPS组相比,黄芩苷干预组H9C2心肌细胞存活率升高(P<0.05),IL-6、IL-1β及TNF-α表达水平降低(P<0.05),p-NF-κB p65、p-p38 MAPK、p-IκBα蛋白水平降低(P<0.05),IκBα蛋白水平升高(P<0.05),而NF-κB p65及p38 MAPK蛋白表达无明显变化(P>0.05)。结论黄芩苷可能通过抑制NF-κB、p38 MAPK的激活来减轻LPS诱导的心肌细胞炎症反应,提高细胞的存活率。Objective To investigate the protective effect of Baicalin on inflammation induced by lipopolysaccharide in H9C2 cardiomyocytes and its possible mechanism. Methods H9C2 myocardial cells were cultured and pretreated with baicalin at the final concentration of 10, 20, 30 μmol/L for 12 hours, then stimulated with LPS at the final concentration of 1 μg/mL for 6 hours. The control group was treated with the same amount of saline to collect cell samples. CCK-8 (The Cell Counting Kit-8) was used to detect cell activity, enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of interleukin-6 (IL-6), interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), Western blot was used to detect the protein expression levels of NF-κB p65, p-NF-κB p65, p38 MAPK, p-p38 MAPK, IκBα and p-IκBα. SPSS 23.0 statistical software was used. Independent sample t test was used for comparison between two groups, and one-way ANOVA test was used for comparison among multiple groups. Results The survival rate of myocardial cells in the control group was (93.67 +1.453)%. Compared with the control group, the survival rate of H9C2 myocardial cells induced by LPS decreased (P < 0.05). In the control group, the expression of IL-6 in H9C2 myocardial cells was (49.33 +2.42) pg/mL, the expression of TNF-α was (86.33 +1.85) pg/mL, and the expression of IL-1β was (28.67 +4.66) pg/mL. Compared with the control group, the expression levels of IL-6, IL-1β and TNF-α in H9C2 myocardial cells increased after LPS induction (P < 0.05), while the levels of p-NF-κ B p65, p-p38 MAPK and p-I κ B α protein increased (P < 0.05), while the levels of I κ B α protein decreased (P < 0.05), while the expressions of NF-κ B p65 and p38 MAPK protein did not change significantly (P > 0.05). Compared with LPS group, the survival rate of H9C2 myocardial cells in baicalin intervention group increased (P < 0.05), the expression levels of IL-6, IL-1β and TNF-a decreased (P < 0.05), the levels of p-NF-κB p65, p-p38 MAPK, p-I κB

关 键 词：黄苓昔 脂多糖 心肌细胞 NF-KB P38MAPK 脓毒症 炎症反应 

分 类 号：R285.5[医药卫生—中药学]