地黄多糖上调miR-216a表达对缺氧/复氧诱导的肾小管上皮细胞氧化应激和细胞凋亡的影响  

Effects of Rehmannia polysaccharide up-regulated miR-216a expression on hypoxia/reoxygenation-induced oxidative stress and apoptosis in renal tubular epithelial cells

作　　者：其其格[1] 夏丽华[1] 任睿 Qi Qige;XIA Lihua;REN Rui(Department of Nephrology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010000,China;Department of Nephrology,Inner Mongolia Traditional Chinese Medicine Hospital,Hohhot 010000,China)

机构地区：[1]内蒙古医科大学附属医院肾内科,内蒙古呼和浩特010000 [2]内蒙古中医医院肾内科,内蒙古呼和浩特010000

出　　处：《沈阳药科大学学报》2023年第11期1466-1472,1576,共8页Journal of Shenyang Pharmaceutical University

基　　金：国家自然科学基金资助项目(81960130)。

摘　　要：目的探讨地黄多糖对缺氧/复氧(H/R)诱导的人肾小管上皮细胞氧化应激和细胞凋亡的影响及可能机制。方法体外培养人肾小管上皮细胞HK-2,用不同剂量(10、20、40μg·mL^(-1))地黄多糖干预HK-2细胞24 h后,建立H/R模型(缺氧6 h,复氧12 h),试剂盒检测细胞中MDA含量及GSH-PX和SOD活性,流式细胞术检测细胞凋亡,蛋白质印迹法检测cleaved-caspase3和cleaved-caspase9蛋白表达,qRT-PCR检测细胞中miR-216a表达。转染miR-216a模拟物或抑制剂至HK-2细胞后,建立H/R模型,上述相同方法检测细胞氧化应激水平和凋亡情况。结果与H/R组比较,地黄多糖降低H/R诱导的HK-2细胞中MDA含量、凋亡率、蛋白(cleaved-caspase3、cleaved-caspase9)表达(P<0.05),而提高GSH-PX和SOD活性(P<0.05)。同时,地黄多糖促进了H/R诱导的HK-2细胞中miR-216a的表达(P<0.05)。上调miR-216a降低H/R诱导的HK-2细胞中MDA含量、凋亡率、蛋白(cleaved-caspase 3、cleaved-caspase 9)表达(P<0.05),而提高GSH-PX和SOD活性(P<0.05)。下调miR-216a逆转地黄多糖对H/R诱导的HK-2细胞氧化应激和凋亡的影响。结论地黄多糖可能通过上调miR-216a抑制H/R诱导的HK-2细胞氧化应激和凋亡。Objective To investigate the effects of polysaccharides from Rehmanniae Radix on hypoxia/reoxygenation(H/R)-induced oxidative stress and apoptosis in human renal tubular epithelial cells.Methods Human renal tubular epithelial HK-2 cells were cultured in vitro.The HK-2 cells were intervened with different doses(10,20,40μg·mL^(-1))of Rehmannia polysaccharide for 24 h,and then the H/R model was established(hypoxia for 6 h,reoxygenation for 12 h).And then the content of MDA and the activities of GSH-PX and SOD in cells were detected by the kit.Cell apoptosis was detected by flow cytometry,and protein expressions of cleaved-caspase3 and cleaved-caspase9 were detected by western blotting,and of miR-216a expression in cells was detected by qRT-PCR.After HK-2 cells were transfected with miR-216a mimics or inhibitors,the H/R model was established,and then the level of oxidative stress and apoptosis of cells were detected by the same method as above.Results Compared with the H/R group,Rehmannia polysaccharide decreased the content of MDA,apoptosis rate and the expression of proteins(cleaved-caspase3,cleaved-caspase9)in HK-2 cells induced by H/R(P<0.05),but increased the activities of GSH-PX and SOD(P<0.05).Meanwhile,Rehmannia polysaccharide promoted the expression of miR-216a in H/R-induced HK-2 cells(P<0.05).Up-regulation of miR-216a decreased the content of MDA,apoptosis rate and the expression of proteins(cleaved-caspase3,cleaved-caspase9)in HK-2 cells induced by H/R(P<0.05),but increased the activities of GSH-PX and SOD(P<0.05).Down-regulation of miR-216a reversed the effects of Rehmannia polysaccharide on H/R-induced oxidative stress and apoptosis in HK-2 cells.Conclusion Rehmannia polysaccharide may inhibit H/R-induced oxidative stress and apoptosis in HK-2 cells by up-regulating miR-216a.

关 键 词：地黄多糖 肾小管上皮细胞 miR-216a 氧化应激 凋亡 

分 类 号：R96[医药卫生—药理学]