枸杞多糖对HK-2细胞缺糖缺氧损伤后线粒体功能及自噬的影响  

Effects of lycium barbarum polysaccharide on mitochondrial function and autophagy of HK-2 cells after hypoglycemia and hypoxia injury

作　　者：严文英 姚靖[2] YAN Wen-ying;YAO Jing(Department of Internal Medicine,Hospital of Xi’an University of Finance and Economics,Xi'an 710100,Shaanxi Province,China;Department of Ophthalmology,The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin,150001,Heilongjiang Province,China)

机构地区：[1]西安财经大学医院内科,陕西西安710100 [2]黑龙江中医药大学附属第一医院眼科,黑龙江哈尔滨150001

出　　处：《中国临床药理学杂志》2020年第24期4010-4012,4017,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81973908)。

摘　　要：目的探讨枸杞多糖对HK-2细胞缺糖缺氧损伤后线粒体功能的影响。方法将细胞分对照组、模型组和实验组(常规培养),模型组(无糖无血清的培养基)和实验组(无糖无血清的培养基+50 mg·L-1枸杞多糖)缺氧培养24 h后复氧培养3 h。以CCK-8法检测细胞存活率,Hoechst 33258核染色法检测细胞凋亡,二氯荧光素二乙酸(DCF-DA)荧光染色检测细胞内活性氧(ROS)水平,JC-1染色法检测细胞线粒体膜电位,分光光度计检测并计算细胞内Ca2+含量,蛋白质免疫印迹法检测细胞自噬蛋白表达情况。结果干预24 h后,对照组、模型组及实验组细胞存活率分别为(98.72±0.14)%,(52.13±4.52)%,(78.68±3.43)%;细胞凋亡率分别为(1.67±0.27)%,(37.24±3.42)%,(16.24±2.14)%;细胞ROS水平分别为0.24±0.02,0.76±0.12,0.47±0.14。模型组分别与对照组和实验组比较,差异均有统计学意义(均P<0.05)。结论枸杞多糖可以抑制缺糖缺氧损伤后的HK-2细胞凋亡,促进其增殖,其机制可能与改善线粒体功能、抑制细胞自噬蛋白表达有关。Objective To investigate the effect of lycium barbarum polysaccharide(LBP) on mitochondrial function of HK-2 cells after glucose-induced hypoxic injury. Methods The cells were divided into control group, model group and experimental group(conventional culture), model group(glucose-free serum-free medium) and experimental group(glucose-free serum-free medium+50 mg-L-1 LBP) were incubated for 24 h in hypoxia and 3 h in reoxygenation. CCK-8 assay was used to detect cell viability. Hoechst 33258 nuclear staining was used to detect cell apoptosis. Intracellular reactive oxygen species(ROS) levels were detected by fluorescence staining with acetic acid(DCF-DA). mitochondrial membrane potential was detected by JC-1 staining. intracellular Ca2+ content was detected and calculated by spectrophotometer, and autophagy protein expression was detected by protein immunoblotting. Results After 24 h of HK-2 cell intervention, the cell survival rates of control, model and test group were(98.72±0.14)%,(52.13±4.52)%,(78.68±3.43)%, respectively;the apoptosis rates were(1.67±0.27)%,(37.24±3.42)%,(16.24±2.14)%, respectively. The ROS levels were 0.24±0.02 0. 76 ± 0. 12, and 0. 47 ± 0. 14, respectively. The differences between the model group and the control and experimental groups were statistically significant(P < 0. 05). Conclusion LBP polysaccharide can inhibit apoptosis and promote the proliferation of HK-2 cells after hypoxic injury,and its mechanism may be related to the improvement of mitochondrial function and inhibition of cellular autophagy protein expression。

关 键 词：枸杞多糖 肾缺血再灌注损伤 缺糖缺氧损伤 线粒体功能 自噬 

分 类 号：R28[医药卫生—中药学]