摘要
目的:研究北柴胡多糖(BCP)体外抗氧化活性,阐明其对D-半乳糖(D-gal)诱导的人肾脏近曲小管HK-2细胞的保护作用。方法:采用热水浸提法提取BCP,检测其抗氧化活性,即体外羟自由基清除率、总抗氧化能力和抗超氧阴离子活性,并与维生素E的体外抗氧化活性进行比较。采用不同浓度BCP和D-gal分别作用于HK-2细胞24 h,采用MTT法检测细胞增殖率,确定BCP和D-gal最适作用浓度。采用D-gal构建人肾脏近曲小管HK-2细胞氧化损伤模型,HK-2细胞分为对照组、D-gal组和不同浓度(25、50、100、200及400 mg·L^(-1))BCP+D-gal组,采用MTT法检测各组细胞增殖率。HK-2细胞分为对照组、D-gal组、BCP+D-gal组和维生素E+D-gal组,采用衰老相关β-半乳糖苷酶(SA-β-gal)染色实验观察各组HK-2细胞SA-β-gal染色情况并计算衰老细胞百分率。结果:抗氧化活性检测,与相同浓度维生素E组比较,不同浓度BCP组羟自由基清除率明显升高(P<0.05),总抗氧化能力明显增强(P<0.05),5、10和15 g·L^(-1) BCP组抗超氧阴离子活性明显增加(P<0.05),且不同浓度BCP组体外抗氧化活性均高于同等浓度维生素E组。与对照组比较,处理HK-2细胞24 h,25~400 mg·L^(-1) BCP组HK-2细胞增殖率呈浓度依赖性升高(P<0.05),且200 mg·L^(-1) BCP组HK-2细胞增殖率最高。与对照组比较,不同浓度D-gal组HK-2细胞增殖率呈时间-浓度依赖性降低,在处理条件为20 g·L^(-1)、48 h时,细胞增殖率降低至对照组的64.77%。与对照组比较,D-gal组HK-2细胞增殖率明显降低(P<0.01);与D-gal组比较,100~400 mg·L^(-1) BCP+D-gal组HK-2细胞增殖率呈浓度依赖性升高(P<0.05或P<0.01)。与对照组比较,D-gal组SA-β-gal染色阳性细胞数明显增多,衰老细胞百分率明显升高(P<0.05);与D-gal组比较,BCP+D-gal组和维生素E+D-gal组SA-β-gal染色阳性细胞数明显减少,衰老细胞百分率明显降低(P<0.05)。结论:BCP具有较强的体外抗氧化活性,可通过抑制细胞
Objective:To study the antioxidant activity in vitro of Bupleurum chinense polysaccharides(BCP),and to clarify its protective effect on D-galactose(D-gal)-induced proximal convolve tubule HK-2 cells of human kidney.Methods:BCP was extracted by hot water extraction method.The antioxidant activities of BCP including the hydroxyl radical scavenging rate,the total antioxidant capacity and the anti-superoxide anion activities were detected,and compared to the antioxidant activities of vitamin E.The HK-2 cells were treated with different concentrations of BCP and D-gal,respectively,and the proliferation rates of HK-2 cells were detected;the optimum concentrations of BCP and D-gal were confinmed.D-gal was used to establish the oxidative damage model of proximal convolve tubule of human kidney.The HK-2 cells were divided into control group,D-gal group,and 25,50,100,200,400 mg·L^(-1) BCP+D-gal groups;MTT method was used to detect the proliferation rates.The HK-2 cells were divided into control group,D-gal group,BCP+D-gal group and vitamin E+D-gal group;senescence-associatedβ-galactosidase(SA-β-Gal)staining was used observe the SA-β-gal staining of the HK-2 cells in various groups and the percentages of senescent the HK-2 groups cells were calculated.Results:The antioxidant activity detection results showed that compared with vitamin E group with the same concentration,the scavenging rates of hydroxyl free radical in different concentrations of BCP groups were increased(P<0.05),and the total antioxidant capacitie were increased(P<0.05);the anti-superoxide anion activities in 5,10,and 15 g·L^(-1) BCP groups were significantly increased(P<0.05),and the antioxidant levels in vitro in different concentrations of BCP groups were higher than those in vitamin E groups with the same concentration.Compared with control group,the proliferation rates of HK-2 cels in 25-400 mg·L^(-1) BCP groups were increased in a concentration-dependent manner after treated for 24 h,and the proliferation rate of HK-2 cells in 200 mg·L^(-1) BCP
作者
侯俊宇
李明慧
许梦然
葛俊宏
申野
李坦
孙新
HOU Junyu;LI Minghui;XU Mengran;GE Junhong;SHEN Ye;LI Tan;SUN Xin(Department of Pathophysiology,School of Basic Medical Science,Beihua University,Jilin 132013,China;Staff Office of Biopharmaceutical,School of Pharmacy,Jilin Medical University,Jilin 132013,China;Department of Social Medicine and Hygiene,School of Public Health,Beihua University,Jilin 132013,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2022年第1期65-73,共9页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅重点实验室项目(20200201178JC)
吉林省发改委自主创新能力项目(2021C007)
吉林医药学院博士基金项目(JYBS2021026LK)
吉林医药学院横向科研项目(2021-HX002)
吉林省吉林市科技局科技创新发展计划项目(201830827)。
关键词
北柴胡多糖
人肾脏近曲小管细胞
氧化损伤
细胞衰老
Bupleurum chinense polysaccharide
Human kidney proximal tubule cells
Oxidative damage
Senescence
