山药多糖对LPS诱导的心肌H9C2细胞炎症因子表达和细胞凋亡的影响及机制

Effect and mechanism of Yam polysaccharide on LPS-induced myocardial H9C2 cell inflammatory factor expression and apoptosis

目的:研究山药多糖对脂多糖(LPS)诱导的心肌H9C2细胞炎症因子表达和细胞凋亡影响及机制。方法:心肌H9C2细胞分成Control组(空白对照)、LPS组(LPS诱导)、CYPS-L组(LPS诱导,0.05 g/L山药多糖处理)、CYPS-M组(LPS诱导,0.10 g/L山药多糖处理)、CYPS-H组(LPS诱导,0.20 g/L山药多糖处理)。CCK-8测定增殖,流式细胞术检测凋亡,荧光法检测ROS水平、黄嘌呤氧化法检测SOD水平、硫代巴比妥酸法检测MDA水平,二硝基苯肼法测定LDH水平,ELISA法检测TNF-α、IL-6水平,Western blot检测C-Caspase-3、p65、IκBα蛋白表达。用NF-κB信号激活剂和山药多糖处理心肌H9C2细胞,同样检测细胞增殖、凋亡和ROS、SOD、MDA、LDH、TNF-α、IL-6水平以及C-Caspase-3、p65、IκBα蛋白表达。结果:与Control组相比,LPS组心肌H9C2细胞增殖活性降低,凋亡率升高,ROS和MDA水平升高,SOD水平降低,培养液上清中LDH、TNF-α、IL-6水平升高,细胞中C-Caspase-3、p65蛋白表达水平升高,IκBα蛋白表达水平下降。与LPS组相比,CYPS-L、CYPS-M、CYPS-H组心肌H9C2细胞增殖活性升高,凋亡率降低,ROS和MDA水平降低,SOD水平升高,培养液上清中LDH、TNF-α、IL-6水平降低,细胞中C-Caspase-3、p65蛋白表达水平降低,IκBα蛋白表达水平升高。NF-κB信号激活剂能够逆转山药多糖对LPS条件下心肌H9C2细胞增殖、凋亡和ROS、SOD、MDA、LDH、TNF-α、IL-6水平以及C-Caspase-3、p65、IκBα蛋白表达影响。结论:山药多糖抑制LPS诱导的心肌H9C2细胞炎症因子表达和细胞凋亡,与降低NF-κB信号激活水平有关。

Objective:To study the effect and mechanism of Yam polysaccharide on lipopolysaccharide(LPS)-induced myocar⁃dial H9C2 cell inflammatory factor expression and apoptosis.Methods:Myocardial H9C2 cells were divided into Control group(blank control),LPS group(LPS induction),CYPS-L group(LPS induction,0.05 g/L Yam polysaccharide treatment),CYPS-M group(LPS induction,0.10 g/L Yam polysaccharide treatment),CYPS-H group(LPS induction,0.20 g/L Yam polysaccharide treat⁃ment).CCK-8 was used to measure proliferation,flow photometry to detect apoptosis,fluorescence to detect ROS level,xanthine oxi⁃dation method to detect SOD level,thiobarbituric acid method to detect MDA level,dinitrophenylhydrazine method to measure LDH level.The levels of TNF-αand IL-6 were detected by ELISA,and the protein expressions of C-Caspase-3,p65 and IκBαwere detected by Western blot.Treated myocardial H9C2 cells with NF-κB signal activator and Yam polysaccharide,and also detect cell prolifera⁃tion,apotheosis and ROS,SOD,MDA,LDH,TNF-α,IL-6 levels and C-Caspase-3,p65,IκBαprotein expression.Results:Com⁃pared with Control group,LPS group myocardial H9C2 cell proliferation activity decreased,apoptosis rate increased,ROS and MDA levels increased,SOD level decreased,LDH,TNF-α,IL-6 levels in culture supernatant increased,the expression levels of C-Cas⁃pase-3 and p65 protein increased,while the expression level of IκBαprotein decreased.Compared with the LPS group,the prolifera⁃tion activity of myocardial H9C2 cells in CYPS-L,CYPS-M,and CYPS-H groups increased,the apoptosis rate decreased,ROS and MDA levels decreased,SOD levels increased,and LDH,TNF-α,IL-6 decreased,the expression levels of C-Caspase-3 and p65 pro⁃tein in cells decreased,and the expression levels of IκBαprotein increased.NF-κB signal activator could reverse the proliferation,apoptosis and ROS,SOD,MDA,LDH,TNF-α,IL-6 levels and C-Caspase-3,p65,IκBαprotein expression of Yam polysaccharide on myocardial H9C2 cells under LPS conditions influences.Conclusion:Yam polysaccharide inhibited LPS-induced myocardial H9C2 cell inflammatory factor expression and apoptosis,the mechanism was related to the reduction of NF-κB signal activation level.