激活SUR2B/Kir6.1亚型K_(ATP)通道对肾脏细胞损伤的干预作用及其机制  

Interventional effects of activating SUR2B/Kir6.1-type K_(ATP)channels on renal cells injury and its mechanisms

作　　者：赵颖[1] 汪海 ZHAO Ying;WANG Hai(Department of Pharmacy,Medical Supplies Center of PLA General Hospital,Beijing 100853;Research Division of Pharmacology,the Fifth Medical Center of PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军总医院医疗保障中心药剂科,北京100853 [2]解放军总医院第五医学中心药理研究室,北京100853

出　　处：《中国应用生理学杂志》2022年第6期604-610,616,共8页Chinese Journal of Applied Physiology

摘　　要：目的:探讨新型SUR2B/Kir6.1亚型K_(ATP)通道开放剂埃他卡林对尿酸所致肾脏细胞(肾小球内皮、系膜和肾小管上皮细胞)损伤干预作用及其机制。方法:①实验分组:对照组(0 mg/L尿酸损伤24 h);模型组(1200 mg/L尿酸损伤24 h);埃他卡林预处理组(终浓度为0.01、0.1、1、10、100μmol/L预孵育24 h+1200 mg/L尿酸损伤24 h);K_(ATP)拮抗组(10μmol/L的格列苯脲孵育1 h后,加入10μmol/L埃他卡林共孵育24 h+1200 mg/L尿酸损伤24 h)。②采用MTT法、流式细胞术检测细胞存活率;免疫荧光检测细胞Kir6.1、SUR2B的蛋白表达及细胞中NF-κB的核转位;Western blot检测细胞Kir6.1、SUR2B的蛋白表达;用荧光强度分析法检测单核细胞对内皮细胞的粘附;用酶联免疫吸附试验(ELISA)检测各组培养液中MCP-1抗原含量。结果:1200 mg/L尿酸作用于肾小球内皮、系膜和肾小管上皮细胞24 h后,与对照组相比,显著降低细胞存活率(P均<0.01)。在肾小球内皮、系膜细胞损伤模型中,埃他卡林0.1、1、10、100μmol/L预处理组与模型组相比,显著升高细胞存活率(P<0.05,P<0.01)。K_(ATP)拮抗剂组与对照组比,细胞存活率显著降低(P<0.01),与模型组比无显著差异(P>0.05),与埃他卡林10μmol/L预处理组比有显著差异(P<0.05,P<0.01)。在肾小管上皮细胞损伤模型中,埃他卡林10、100μmol/L预处理组与模型组相比,显著升高细胞存活率(P<0.05)。K_(ATP)拮抗剂组与对照组比,细胞存活率显著降低(P<0.01),与模型组比无显著差异(P>0.05)。肾小球内皮、系膜和肾小管上皮细胞中,模型组与对照组相比,显著升高Kir6.1、SUR2B的蛋白表达(P均<0.05)。100x0E䥺SymbolmA@0x0Fmol/L埃他卡林预处理组与模型组相比,显著抑制上调Kir6.1、SUR2B蛋白的表达(P均<0.05)。K_(ATP)拮抗剂组Kir6.1、SUR2B的蛋白表达升高,与模型组比均无显著差异(P>0.05)。1200 mg/L尿酸与肾小球内皮细胞孵育24 h后,与对照组相比,单核细胞对内皮细�Objective:To investigate the interventional effects of a new SUR2B/Kir6.1-type K_(ATP)Channel opener iptakalim on injury renal cells(the renal glomerular endothelial,mesangial and tubular epithelial cells)and its mechanisms.Methods:①Experimental protocol:control:the cells were treated with with 0 mg/L uric acid for 24 h;model:the cells were treated with with 1200 mg/L uric acid for 24 h;pretreatment with iptakalim:the cells were pretreated with 0.01,0.1,1,10,100μmol/L iptakalim for 24 h prior to treatment with 1200 mg/L uric acid for 24 h;pretreatment with glibenclamide:the cells were preincubated with/without 10μmol/L glibenclamide for 1 h and then treated with 10μmol/L iptakalim for 24 h followed by incubation with 1200 mg/L uric acid for another 24 h.②The cell viability was measured by MTT assay and flow cytometry;the protein expressions of Kir6.1 and SUR2B and nuclear translocation were detected by immunostaining;the protein expressions of Kir6.1 and SUR2B were determined by Western blot analysis;adhesion of mononuclear cells to endothelial cells were tested by fluorimetric assay;the content of MCP-1 was measured by enzyme linked-immunosorbent assay(ELISA).Results:The renal glomerular endothelial,mesangial and tubular epithelial cells were exposed to 1200 mg/L uric acid for 24 h.Compared with the control group,1200 mg/L uric acid decreased the cell survival rates significantly(P<0.01,P<0.01,P<0.01).Compared with the model group,pretreatment with 0.1,1,10,100μmol/L iptakalim could remarkably alleviate cellular damages of glomerular endothelium,mesangium cells induced by uric acid(P<0.05,P<0.01,P<0.01,P<0.01).The K_(ATP)channel blocker could clearly reduce survival rates of the renal glomerular endothelial,mesangial cells(P<0.01)and markedly reverse the inhibitory effects of iptakalim on cell death(P<0.05,P<0.01),no obvious difference in comparison with the model group(P>0.05).Compared with the model group,pretreatment with 10,100μmol/L iptakalim could notably attenuate cellular damages of tubular epit

关 键 词：SUR2B/Kir 6.1亚型 ATP敏感性钾离子通道 埃他卡林 肾脏损伤 高尿酸血症 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学]