GLUT1依赖性的糖酵解在右美托咪定减轻HK-2细胞氧糖剥夺-复氧复糖损伤中的作用  

Role of GLUT1-dependent glycolysis in attenuation of oxygen-glucosedeprivation-reoxygenation injury by dexmedetomidine in HK-2 cells

作　　者：丁威 陶文辉 吴雨乐 吴剑霄 郭婧怡 谢丽芳 樊炳乾 谷雪松 李洋[4] 胡宪文[1] DING Wei;TAO Wen-hui;WU Yu-le;WU Jian-xiao;GUO Jing-yi;XIE Li-fang;FAN Bing-qian;GU Xue-song;LI Yang;HU Xian-wen(Dept of Anesthesiology and Perioperative Medicine,the Second Hospital of Anhui Medical University,Key Laboratory of Anesthesiology and Perioperative Medicine of Anhui Higher Education Institutes,Hefei 230601,China;the First Clinical College,Anhui Medical University,Hefei 230032,China;the Second Clinical College,Anhui Medical University,Hefei 230601,China;Dept of Genetics,School of Life Sciences,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学第二附属医院麻醉与围术期学科,麻醉与围术期医学安徽普通高校重点实验室,安徽合肥230601 [2]安徽医科大学第一临床医学院,安徽合肥230032 [3]安徽医科大学第二临床医学院,安徽合肥230601 [4]安徽医科大学生命科学院遗传学系,安徽合肥230032

出　　处：《中国药理学通报》2025年第3期444-450,共7页Chinese Pharmacological Bulletin

基　　金：安徽省高校科研重点项目(No 2023AH053181);2023年临床医学学科建设项目(No 9101059801);安徽医科大学第二附属医院国家自然科学基金孵育计划(No 2023GMFY03);2023年度临床医学(“5+3”一体化)专业“早期接触科研”训练计划项目(No 2023-ZQKY-017);安徽医科大学校基金资助项目(No 2023xkj026,2023xkj030)。

摘　　要：目的评价葡萄糖转运蛋白1(glucose transporter 1,GLUT1)依赖性的糖酵解在右美托咪定(dexmedetomidine,Dex)减轻人肾小管上皮(human kidney-2,HK-2)细胞氧糖剥夺-复氧复糖(oxygen-glucose deprivation-reoxygenation,OGD/R)损伤中的作用。方法将C57/BL6小鼠随机分为3组(n=6):假手术组(Sham组),肾缺血再灌注组(I/R组),Dex组(I/R+Dex组)。检测肌酐(Cr)和尿素氮(BUN),GLUT1和糖酵解关键酶HK2、PFKFB3蛋白水平。HK-2细胞随机分为7组(n=6),对细胞进行OGD/R,过表达或干扰GLUT1,以及Dex、2-DG等处理,通过CCK-8、LDH反映细胞损伤程度,乳酸和细胞外酸化率(ECAR)评估糖酵解水平,qRT-PCR检测IL-6和TNF-α反映炎症水平。qRT-PCR和Western blot检测GLUT1、HK2、PFKFB3水平。结果Dex改善了组织和细胞损伤(P<0.05),抑制了OGD/R诱发的乳酸和ECAR上升以及GLUT1、HK2、PFKFB3的高表达(P<0.05)。体外实验表明GLUT1敲低后改善了OGD/R诱发的HK-2细胞损伤,降低了乳酸和ECAR水平,GLUT1、HK2、PFKFB3的mRNA和蛋白表达下降(P<0.05)。而在敲低GLUT1基础上加用Dex处理后对以上指标均无影响。过表达GLUT1能消除Dex的保护作用,逆转Dex对GLUT1、HK2、PFKFB3的抑制作用(P<0.05)。结论Dex通过抑制GLUT1依赖性糖酵解减轻了OGD/R诱发的HK-2细胞损伤。Aim To evaluate the role of the glucose transporter protein 1(GLUT1)-dependent glycolytic in the attenuation of oxygen-glucose deprivation-reoxygenation(OGD/R)injury in HK-2 cells by dexmedetomidine(Dex).Methods C57/BL6 mice were randomly divided into three groups(n=6),namely,sham operation group(Sham group),renal ischemia reperfusion group(I/R group)and Dex group(I/R+Dex group).Serum creatinine(Cr)and urea nitrogen(BUN)were measured,while the levels of key glycolytic enzymes HK2,PFKFB3 and GLUT1 were measured.HK-2 cells were cultured and randomised into seven groups(n=6),which was treated with OGD/R,overexpression or interference with GLUT1,Dex and glycolysis inhibitor 2-DG.CCK-8 and LDH activity were used to detect cellular damage.Glycolysis levels were detected by lactate and ECAR.The inflammatory level was reflected by qRT-PCR for IL-6 and TNF-α.qRT-PCR and Western blot were performed to detect the levels of GLUT1,HK2,and PFKFB3.Results Dex significantly ameliorated kidney injury and HK-2 cell injury(P<0.05).Dex inhibited the OGD/R-induced rise in lactate and extracellular acidification rate(ECAR),as evidenced by suppression of the expression of GLUT1,HK2 and PFKFB3(P<0.05).In vitro experiments showed that GLUT1 knockdown significantly improved OGD/R-induced cellular damage.Lactate,ECAR,glycolysis-related mRNAs and proteins were inhibited by GLUT1 knockdown(P<0.05).Significantly,there were no significant differences in above indexes after Dex treatment based on GLUT1 knockdown.Overexpression of GLUT1 abrogated the protective effects of Dex,while reversing the inhibitory effects of Dex on the expression of GLUT1,HK2,and PFKFB3(P<0.05).Conclusions Dexmedetomidine attenuates OGD/R induced injury in HK-2 cells by inhibiting GLUT1-dependent glycolysis.

关 键 词：葡萄糖转运蛋白1 氧糖剥夺-复氧复糖 糖酵解 右美托咪定 肾小管上皮细胞 缺血/再灌注 

分 类 号：R-332[医药卫生] R322.61R341R343R614R852.15R692.5