雷公藤多苷对高糖诱导人肾小管上皮细胞凋亡及CXCL10/CXCR3轴的影响  

Effects of tripterygium glycosides on high glucose induced apoptosis of human renal tubular epithelial cells and CXCL10/CXCR3 axis

作　　者：李明霞[1] 乔海霞[2] 王晓玲[1] 贾丽媛[1] 胡利梅[1] 任卫东[1] LI Mingxia;QIAO Haixia;WANG Xiaoling;JIA Liyuan;HU Limei;REN Weidong(Department of Endocrinology,the First Hospital Affiliated to Hebei North University,Zhangjiakou 075000,China;Clinical Medical College of Hebei North University)

机构地区：[1]河北北方学院附属第一医院内分泌科,075000 [2]河北北方学院临床医学院

出　　处：《天津医药》2023年第2期139-143,共5页Tianjin Medical Journal

基　　金：河北省医学科学研究课题计划项目(20190871)。

摘　　要：目的 探讨雷公藤多苷(TG)对高糖诱导人肾小管上皮细胞HK-2凋亡及CXC趋化因子配体10(CXCL10)/CXC趋化因子受体3(CXCR3)轴的影响。方法 体外培养人肾小管上皮细胞HK-2,并分为对照组(含5.5 mmol/L葡萄糖培养基)、高糖组(含25 mmol/L葡萄糖培养基)和12.5、25、50 mg/L TG组(分别用12.5、25、50 mg/L的TG和25 mmol/L葡萄糖培养基)。四甲基偶氮唑盐比色法检测HK-2细胞活力;流式细胞术检测HK-2细胞凋亡情况;2’,7’-二氯二氢荧光素二乙酸酯法检测HK-2细胞活性氧(ROS)水平;黄嘌呤氧化酶法检测HK-2细胞超氧化物歧化酶(SOD)水平;酶联免疫吸附试验检测HK-2细胞炎性因子肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)水平;蛋白免疫印迹法检测HK-2细胞凋亡蛋白[胱天蛋白酶(Caspase)-3、Caspase-9]以及CXCL10、CXCR3蛋白表达。结果 与对照组比较,高糖组HK-2细胞活力、SOD水平显著降低,凋亡率、ROS、TNF-α、TGF-β1水平及Caspase-3、Caspase-9、CXCL10、CXCR3蛋白表达升高(P<0.05);与高糖组比较,12.5、25、50 mg/L TG组HK-2细胞活力、SOD水平升高,凋亡率、ROS、TNF-α、TGF-β1水平及Caspase-3、Caspase-9、CXCL10、CXCR3蛋白表达降低,且高剂量效果更好(P<0.05)。结论 TG可抑制高糖诱导的HK-2细胞凋亡、氧化应激和炎症反应,其机制可能与抑制CXCL10/CXCR3轴有关。Objective To investigate effects of tripterygium glycosides(TG) on the apoptosis of human renal tubular epithelial cells HK-2 induced by high glucose and the CXC chemokine ligand 10(CXCL10)/CXC chemokine receptor 3(CXCR3) axis.Methods Human renal tubular epithelial cells HK-2 were cultured in vitro and divided into the control group(culture medium containing 5.5 mmol/L glucose),the high glucose group(culture medium containing 25 mmol/L glucose) and the 12.5,25 and 50 mg/L TG groups(culture medium containing 12.5,25 and 50 mg/L TG and 25 mmol/L glucose,respectively).Methyl thiazolyl tetrazolium(MTT) method was used to detect the viability of HK-2 cells.Flow cytometry was used to detect the apoptosis of HK-2 cells.2’,7’-dichlorodi-hydrofluorescein diacetate method was used to detect reactive oxygen species(ROS) level of HK-2 cells.Xanthine oxidase method was used to detect superoxide dismutase(SOD) level of HK-2 cells.Enzme-linked immunosorbent assay was used to detect levels of inflammatory factors of tumor necrosis factor-α(TNF-α) and transforming growth factor-β1(TGF-β1) in HK-2 cells.Western blot assay was used to detect the expression of apoptotic proteins(Caspase-3,Caspase-9),CXCL10 and CXCR3 proteins in HK-2 cells.Results Compared with the control group,the HK-2 cell viability and SOD level were significantly reduced in the high glucose group,and the apoptosis rate,ROS,TNF-α,TGF-β1 levels and Caspase-3,Caspase-9,CXCL10,CXCR3 protein expression levels were significantly increased(P <0.05).Compared with the high glucose group,the HK-2 cell viability and SOD level were significantly increased in the 12.5,25 and 50 mg/L TG groups,and the apoptosis rate,ROS,TNF-α,TGF-β1 levels and Caspase-3,Caspase-9,CXCL10,CXCR3 protein expression levels were significantly reduced,and high dose TG has better effect(P <0.05).Conclusion TG can inhibit high glucose induced apoptosis,oxidative stress and inflammation of HK-2 cells,and its mechanism may be related to the inhibition of CXCL10/CXCR3 axis.

关 键 词：雷公藤 趋化因子CXCL10 受体 CXCR3 细胞凋亡 氧化性应激 雷公藤多苷 高糖 人肾小管上皮细胞HK-2 

分 类 号：R587.24[医药卫生—内分泌]