摘要
目的:观察铁皮石斛多糖(DP)对心肌细胞缺血再灌注损伤的保护作用并探讨其作用机制。方法:建立H9c2大鼠心肌细胞缺氧/复氧模型,将H9c2细胞分为正常对照(Con)组,缺氧/复氧(H/R)组(缺氧3 h,复氧3 h),缺氧/复氧+铁皮石斛多糖低(H/R+LDP 0.1μg/L)、中(H/R+MDP 0.2μg/L)、高(H/R+HDP 0.4μg/L)质量浓度组(DP预处理30 min后行H/R处理)。MTT法检测各组H9c2细胞存活率,比色法检测细胞内ROS水平、MDA含量和上清液中LDH含量,AnnexinⅤ-FITC/PI双染法检测细胞凋亡率,Western blot法检测NF-κB p65和TLR-4蛋白表达水平。结果:与Con组比较,H/R组H9c2细胞存活率降低,细胞中ROS水平及MDA含量升高,上清液中LDH含量增加,细胞凋亡率升高,细胞中NF-κB p65和TLR-4蛋白的表达水平上调(P<0.05);与H/R组比较,铁皮石斛多糖预处理后,H9c2细胞存活率升高,细胞中ROS水平及MDA含量减少,上清液中LDH含量减少,细胞凋亡率降低,细胞中NF-κB p65和TLR-4蛋白的表达水平下调(P<0.05)。结论:铁皮石斛多糖对H9c2细胞缺氧/复氧损伤具有一定的保护作用,其作用机制可能与提高细胞活性、增强抗氧化损伤能力、抑制细胞凋亡、抑制NF-κB/TLR-4信号通路有关。
Aim:To observe the protective effects of Dendrobium candidum polysaccharide(DP)on hypoxia/reoxygenation(H/R)of cardiomyocytes and its mechanism.Methods:H/R model of H9c2 rat cardiomyocytes were established.The cardiomyocytes were divided into normal control(Con)group,H/R group(hypoxia 3 hours,reoxygenation 3 hours),H/R+DP low(H/R+LDP 0.1μg/L),medium(H/R+MDP 0.2μg/L),high(H/R+HDP 0.4μg/L)concentration groups.The survival rate of cardiomyocytes in each group was detected by MTT assay.Intracellular ROS level,MDA level,as well as LDH level in supernatant were detected using colorimetric assay.Apoptosis rate was detected by Annexin V-FITC/PI double staining.NF-κB p65 and TLR-4 protein expression levels were detected using Western blot.Results:Compared with the Con group,the survival rate of cardiomyocytes in the H/R group decreased,the levels of ROS and MDA in the cells increased,the LDH level in supernatant increased,the apoptosis rate increased,and the expression levels of NF-κB p65 and TLR-4 were up-regulated in the cells(P<0.05).Compared with the H/R group,the pretreatment of DP increased the survival rate of cardiomyocyte,as well as decreased the levels of ROS and MDA in the cells,decreased the LDH level in supernatant,as well as decreased the apoptosis rate and the expression levels of NF-κB p65 and TLR-4(P<0.05).Conclusion:DP has protective effects on H/R injury of cardiomyocytes,and its mechanism may be related to increasing cell activity,enhancing its anti-oxidative damage ability,inhibiting apoptosis and inhibiting NF-κB/TLR-4 signaling pathway.
作者
徐姗
胡楠
XU Shan;HU Nan(Department of Pharmacy,the Affiliated Changzhou Second People′s Hospital,Nanjing Medical University,Changzhou,Jiangsu 213000;Department of Pharmacy,Changzhou First People′s Hospital,Changzhou,Jiangsu 213000)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2019年第5期766-769,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
江苏省药学会天晴医院药学基金项目(Q2018105)
