黄芩苷对酯多糖损伤AC16心肌细胞的保护作用及机制研究
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摘要
本研究主要探讨黄芩苷对酯多糖损伤AC16心肌细胞的保护作用。设置对照组、LPS组、黄芩苷高、中、低剂量组、黄芪总黄酮组,共同培养6 h,检测细胞生存率及生存状况;荧光素法检测细胞内ATP浓度变化; Western blot检测各组细胞中Cyt C、Apaf-1、caspase-3、caspase-9表达。结果发现与LPS组相比,黄芩苷高、中、低剂量组及黄芪总黄酮组细胞生存状况及生存率明显改善,细胞内ATP浓度升高,凋亡率降低,细胞内Cyt C、Apaf-1、caspase-3及caspase-9表达均降低。说明黄芩苷对酯多糖损伤的AC16心肌细胞具有保护作用,其机制可能与降低Cyt C/Apaf-1/caspase-3/caspase-9通路表达有关。
This study mainly discussed the protective effect of baicalin on AC16 cardiomyocytes injured by Lipopolysaccharide.The experiment was divided into control group,LPS group,baicalin high,medium and low dose groups,total flavonoids of astragalus membranaceus group and cultured for 6 hours to test cell survival rate and survival status; The firefly luciferin method detects changes in intracellular ATP concentration; The expressions of Cyt C,Apaf-1,caspase-3 and caspase-9 in each group were detected by Western blot.The results showed that compared with the LPS group,the cell survival rate and survival status of the high,middle and low doses group of baicalin and the total flavonoids of astragalus membranaceus group were significantly improved,the intracellular ATP concentration increased,and the apoptotic rate decreased.The expression of Cyt C,Apaf-1,caspase-3 and caspase-9 was decreased in cells.It indicated that baicalin has protective effect on AC16 cardiomyocytes damaged by LPS,and its mechanism may be related to the decrease of Cyt C/Apaf-1/caspase-3/caspase-9 pathway expression.
This study mainly discussed the protective effect of baicalin on AC16 cardiomyocytes injured by Lipopolysaccharide.The experiment was divided into control group,LPS group,baicalin high,medium and low dose groups,total flavonoids of astragalus membranaceus group and cultured for 6 hours to test cell survival rate and survival status; The firefly luciferin method detects changes in intracellular ATP concentration; The expressions of Cyt C,Apaf-1,caspase-3 and caspase-9 in each group were detected by Western blot.The results showed that compared with the LPS group,the cell survival rate and survival status of the high,middle and low doses group of baicalin and the total flavonoids of astragalus membranaceus group were significantly improved,the intracellular ATP concentration increased,and the apoptotic rate decreased.The expression of Cyt C,Apaf-1,caspase-3 and caspase-9 was decreased in cells.It indicated that baicalin has protective effect on AC16 cardiomyocytes damaged by LPS,and its mechanism may be related to the decrease of Cyt C/Apaf-1/caspase-3/caspase-9 pathway expression.
引文
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2 Esper AM,Martin GS.Extending international sepsis epidemiology:the impact of organ dysfunction[J].Crit Care,2009,13:120-120.
3 Li X,Li JH.Influencing factors and research progress of sepsis animal models[J].Chin J Cri Car Med(中华危重症医学杂志),2015,8:53-57.
4 Long W,Mu GN,Zhang GH,et al.Dark tea extracts protect mice from sepsis[J].Nat Prod Res Dev(天然产物研究与开发),2018,30:582-587.
5 Smeding L,Plotz FB,Groeneveld AB,et al.Structural changes of the heart during severe sepsis or septic shock[J].Shock,2012,37:449-456.
6 Pei Z,Wang B,Zhang F,et al.Response of human periodontal ligament cells to baicalin[J].J Periodontol,2014,85:1283-1290.
7 Wu Q,Ye H,Zhu YZ,et al.Protective effect of baicalin against LPS-induced intestinal injury[J].Chin J Chin Mater Med(中国中药杂志),2013,24:2854-2857.
8 He LM,Wang H,Zhao HY,et al.Effects of baicalin on blood pressure and left ventricular remodeling in rats with renal hypertension[J].Chin J Arterioscler(中国动脉硬化杂志),2017,25:693-700.
9 Luan Y,Zheng M,Lu Y,et al.Protective effect of baicalin on the culture hydroxide injury of Cardiac myocyte[J].JJinzhou Med College(锦州医学院学报),2006,27:14-17.
10 Yang L,Shen MZ,Wang B,et al.Effect of baicalin on tunicamycin-induced endoplasmic reticulum stress injury in cultured neonatal rat cardiomyocytes[J].Chin Heart J(心脏杂志),2014,26:125-128.
11 Wang S,Hu Y,Bao ST.Effect of Baicalin on ventricular remodeling,ventricular myocyte apoptosis andβ-AR/PKA/Ca MKⅡsignaling pathway in dilated cardiomyopath rats[J].Chin J Exp Trad Med Form(中国实验方剂学杂志),2018,24:1-5.
12 Wang HJ,Yu M,Wang YH,et al.Effect of total flavonoids of astragalus on endoplasmic reticulum chaperone,apoptosis signaling factor in mouse myocardium with myocarditis caused by coxsackievirus B3[J].J Clin Card(临床心血管病杂志),2016,32:70-73.
13 Xu Y,Chen GB,Su J,et al.Expression of ET-1,NO and E-selectin and the protective effect of milrinone on myocardial injury in sepsis rats in early stage[J].Acta Med Univ Sci Technol Huazhong(华中科技大学学报:医学版),2013,42:195-199.
14 Kurt AN,Aygun AD,Godekmerdan A,et al.Serum IL-1 beta,IL-6,IL-8,and TNF-alpha levels in early diagnosis and management of neonatal sepsis[J].Mediators Inflamm,2007,2007:31397.
15 Chu YG,Qi HN,Zhang JJ,et al.Effect of Shuxuening on early myocardial injury markers and oxidative stress in severe sepsis patients with myocardial injury[J].Acad J Second Military Med Univ(第二军医大学学报),2017,38:804-809.
16 Zhu Y,Yan J.Role of calcium imbalance in myocardial injury in sepsis[J].Pre Tre Car-Cer-Vas Dis(心脑血管病防治),2014,14:149-151.
17 Li DH,Hu DL,Li YN,et al.Correlation between inflammatory cytokines and myocardial injury in early stages of sepsis[J].Anhui Med J,2014,35:1627-1629.
18 Wang P,Ma JJ,Du YM.Protective effect of Baicalin on myocardial ischemia reperfusion injury and its correlation to myocardial cell autophagy in experimental rats[J].Chin Cir J(中国循环杂志),2016,31:701-705.
19 Bai J,Zhang WL,Zhang JW,et al.Myocardial oxidative stress injury and myocardial ultrastructure in septic rats[J].Chin J Cell Mol Immunol(细胞与分子免疫学杂志),2015,31:634-638.
20 Brunelle JK,Letai A.Control of mitochondrial apoptosis by the Bcl-2 family[J].J Cell Sci,2009,122:437-441.