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Received:January 16, 2020 Published Online:August 20, 2020
Received:January 16, 2020 Published Online:August 20, 2020
中文摘要: 目的 观察利拉鲁肽(LIR)对波动高糖诱导的人髓核细胞氧化应激和炎性损伤的保护作用。方法 培养人髓核细胞株,第三代髓核细胞随机分为7组:(1)5.5 mmol/L低浓度葡萄糖对照组(CON组);(2)33 mmol/L持续高浓度葡萄糖组(H组);(3)5.5～33 mmol/L波动高糖组(F组);(4)33 mmol/L高渗对照组(P组);(5)LIR 10 nmol/L(nM)干预组(LIR 10组):5.5～33 mmol/L波动葡萄糖+10 nM LIR;(6)LIR 100 nM干预组(LIR 100组):5.5～33 mmol/L波动葡萄糖+100 nM LIR;(7) LIR 1 000 nM干预组(LIR 1 000组):5.5～33 mmol/L波动葡萄糖+1 000 nM LIR。各组细胞培养72 h,CCK-8对人髓核细胞的增殖活性进行定量分析,流式细胞术检测细胞内活性氧(ROS)水平,ELISA检测各组细胞上清液中丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、肿瘤坏死因子α(TNF-α)与细胞间黏附分子-1(ICAM-1)的含量。结果 与CON组和P组相比,H组、F组、LIR干预组(LIR 10、100、1 000 nM)的细胞增殖活性（OD值）、SOD、GSH降低,ROS水平及MDA、TNF-α、ICAM-1含量增加,差异有统计学意义(P<0.05);P组与CON组相比各指标差异均无统计学意义(P>0.05);F组与H组相比,细胞增殖活性、SOD、GSH水平明显下降,MDA、ROS、ICAM-1与TNF-α含量升高(P<0.05)。与F组相比,LIR各浓度干预组细胞增殖活性、SOD活性、GSH含量均增加,ROS、MDA、ICAM-1与TNF-α含量均下降(P<0.05),且LIR在中等浓度(100 nM)达到最大作用效果。结论 LIR可能通过减轻波动高糖诱导的人髓核细胞的氧化应激及炎性反应,从而发挥保护作用。
Abstract:ObjectiveTo observe the protective effect of liraglutide (LIR) on oxidative stress and inflammatory injury of human nucleus pulposus cells(NPCs) induced by fluctuant high glucose.MethodsAfter NPCs lines were cultured,the 3rd generation of NPCs randomly divided into the following 7 groups:(1) 5.5 mmol/L low concentration glucose control group (CON group);(2) 33 mmol/L sustained high concentration glucose group (H group);(3) 5.5～33 mmol/L fluctuant high glucose group (F group);(4) 33 mmol/L hyperosmolar control group (P group);(5) LIR 10 nmol/L (nM) intervention group (LIR 10 group):5.5～33 mmol/L fluctuant glucose+10 nM LIR;(6) LIR 100 intervention group (LIR 100 group):5.5～33 mmol/L fluctuant glucose+100 nM LIR;(7) LIR 1 000 intervention group (LIR 1 000 group):5.5～33 mmol/L fluctuant glucose+1 000 nM LIR.After 72 hours of cell culture in each group,CCK-8 was used to analyze the proliferation activity of NPCs.Flow cytometry was used to detect intracellular reactive oxygen species (ROS) levels.ELISA was used to detect the malondialdehyde (MDA),superoxide dismutase (SOD),reduced glutathione (GSH),tumor necrosis factor alpha (TNF-α) and intercellular adhesion molecule-1 (ICAM-1) content.ResultsCompared with the CON group and P group,the cell proliferation activity(OD),SOD,GSH levels decreased,ROS level and MDA,TNF-α,ICAM-1 levels increased in H group,F group,LIR intervention group (LIR 10,100,1 000 nM)(P<0.05);there was no statistically significant difference in the above indicators between the P group and the CON group (P>0.05);compared with H group,cell proliferation activity,SOD,and GSH levels decreased,and MDA,ROS,ICAM-1,TNF-α increased in F group(P<0.05).Compared with F group,the cell proliferation activity,SOD activity,and GSH content increased,and the ROS,MDA,ICAM-1,TNF-α contents decreased in each LIR intervention group (P<0.05),and the LIR achieved its maximum effect at medium concentration (100 nM).ConclusionLIR may play a protective role by reducing the oxidative stress and inflammatory response of human NPCs induced by fluctuant high glucose.
keywords: Liraglutide Human nucleus pulposus cells Fluctuation High glucose Oxidative stress Inflammatory injury
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基金项目:国家自然科学基金(31701238)
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