铁皮石斛多糖减轻高糖诱导的HK-2细胞损伤研究  

Study on the inhibitory effect of Dendrobium candidum polysaccharides on high glucose-induced HK-2 cells injury

作　　者：韩秀平[1] 谭娟[2] 贺钰梅[1] Han Xiuping;Tan Juan;He Yumei(Department of General Medicine,The Affiliated Hospital of Yan’an University,Yan'an 716000,China;Internal Medicine of Traditional Chinese Medicine,The Affiliated Hospital of Yan’an University,Yan'an 716000,China)

机构地区：[1]延安大学附属医院全科医学科,延安716000 [2]延安大学附属医院中医内科,延安716000

出　　处：《广西医科大学学报》2021年第11期2121-2127,共7页Journal of Guangxi Medical University

基　　金：延安市科学技术局资助项目(No.2018CGZH-15)。

摘　　要：目的:探讨铁皮石斛多糖是否通过调控微小RNA(miR)-363来影响高糖诱导的人肾小管上皮细胞HK-2损伤。方法:将HK-2细胞分为NG组(正常糖,5 mmol/L葡萄糖),HG组(高糖,30 mmol/L葡萄糖),HG+铁皮石斛多糖-L、M、H组(30 mmol/L葡萄糖+100μg/mL、200μg/mL、400μg/mL铁皮石斛多糖),HG+miR-NC(30 mmol/L葡萄糖+转染miR-NC)、HG+miR-363组(30 mmol/L葡萄糖+转染miR-363模拟物)、HG+铁皮石斛多糖+anti-miR-363组(30 mmol/L葡萄糖+转染anti-miR-363+400μg/mL铁皮石斛多糖)。采用细胞计数试剂盒8(CCK-8)法检测细胞存活率;流式细胞术检测细胞凋亡;western blotting检测Bax、Bcl-2表达;试剂盒检测氧化应激指标超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;定量Real-time PCR检测miR-363的表达。结果:铁皮石斛多糖提高高糖诱导的HK-2细胞的存活率、Bcl-2蛋白水平、SOD活性、miR-363表达水平,降低凋亡率、Bax蛋白水平、MDA含量,且均呈浓度依赖性(P<0.05)。过表达miR-363使高糖诱导的HK-2细胞的活性、Bcl-2蛋白水平、SOD活性、miR-363表达水平提高,凋亡率、Bax蛋白水平、MDA含量减少(P<0.05)。铁皮石斛多糖对高糖诱导的HK-2细胞存活率、凋亡及氧化应激的影响可被抑制miR-363逆转。结论:铁皮石斛多糖通过促进miR-363表达,增强高糖诱导的HK-2细胞活性,减弱其凋亡和氧化应激。Objective:To investigate the effect of Dendrobium candidum polysaccharide(DCP)regulating microRNA(miR)-363 on the human renal tubular epithelial HK-2 cells injury induced by high glucose(HG).Methods:The HK-2 cells were divided into normal glucose(NG)group(5 mmol/L glucose),HG group(30 mmol/L glucose),HG+DCP low-dose(DCP-L)group(30 mmol/L glucose+100μg/mL DCP),HG+DCP medium-dose group(30 mmol/L glucose+200μg/mL DCP),HG+DCP high-dose group(30 mmol/L glucose+400μg/mL DCP),HG+miR-NC group(30 mmol/L glucose+miR-NC transfection),HG+miR-363 group(30 mmol/L glucose+miR-363 mimic transfection),and HG+DCP+anti-miR-363 group(30 mmol/L glucose+400μg/mL DCP+anti-miR-363 transfection).CCK-8 assay was used to detect cell survival rate.Flow cytometry was employed to measure cell apoptosis.Western blotting was used to determine the expressions of Bax and Bcl-2.The superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were detected as well.The expression of miR-363 was determined by real-time PCR.Results:DCP led to dose-dependent increases in the survival rate of HK-2 cells induced by HG,Bcl-2 protein level,SOD activity and miR-363 expression level,and decreases in the cell apoptosis rate,Bax protein level and MDA content(P<0.05).Overexpression of miR-363 by transfection with miR-363 mimic increased the survival rate of HG-induced HK-2 cells,Bcl-2 protein level,SOD activity and miR-363 expression level,while decreased cell apoptosis rate,Bax protein level and MDA content(P<0.05).The effects of DCP on the proliferation,apoptosis and oxidative stress of HK-2 cells were reversed by anti-miR-363.Conclusion:DCP can enhance the activity of HK-2 cells induced by HG through promoting the expression of miR-363,and attenuating cell apoptosis and oxidative stress.

关 键 词：糖尿病肾病 铁皮石斛多糖 miR-363 HK-2细胞 凋亡 氧化应激 

分 类 号：R587.1[医药卫生—内分泌]