欧前胡素调节HMGB1-RAGE-NF-κB信号通路对高糖诱导的视网膜内皮细胞损伤的影响  

Effect of Imperatorin on High Glucose Induced Retinal Endothelial Cell Injury by Regulating the HMGB1-RAGE-NF-κB Signaling Pathway

作　　者：马静[1] 王齐[2] 陈彩玲[3] 谭丹丹[3] 刘延东[4] 王微微[3] MA Jing;WANG Qi;CHEN Cailing;TAN Dandan;LIU Yandong;WANG Weiwei(Nursing Teaching and Research Office,Heilongjiang Provincial Hospital,Harbin 150036,China;Department of Hospital Ofice,Heilongjiang Provincial Hospital,Harbin 150036,China;Department of Ophthalmology,Heilongjiang Provincial Hospital,Harbin 150036,China;Heilongjiang Provincial HHoosspnitaitla Ilntegrated Traditional Chinese and Western Medicine Medical Center,Harbinn 150036,China)

机构地区：[1]黑龙江省医院护理教研室,哈尔滨150036 [2]黑龙江省医院院办公室,哈尔滨150036 [3]黑龙江省医院眼科,哈尔滨150036 [4]黑龙江省医院中西医结合医疗中心,哈尔滨150036

出　　处：《中国细胞生物学学报》2024年第8期1506-1514,共9页Chinese Journal of Cell Biology

基　　金：黑龙江省卫生健康委科研课题(批准号:20231414050307)资助的课题。

摘　　要：该研究探讨欧前胡素(IMP)调节高迁移率族蛋白B1(HMGB1)-晚期糖基化终产物受体(RAGE)-核转录因子κB(NF-κB)信号通路对高糖诱导的视网膜血管内皮细胞损伤的影响。使用10~320μmol/L的IMP处理高糖诱导的视网膜血管内皮细胞hRECs,筛选最佳药物浓度;将视网膜血管内皮细胞hRECs细胞分为正常糖组(NG组,5.5 mmol/L葡萄糖)、高糖组(HG组,25 mmol/L葡萄糖)、低浓度欧前胡素组(IMP-L组,25 mmol/L葡萄糖+20μmol/L IMP)、中浓度欧前胡素组(IMP-M组,25 mmol/L葡萄糖+40μmol/L IMP)、高浓度欧前胡素组(IMP-H组,25 mmol/L葡萄糖+80μmol/L IMP)、高浓度欧前胡素+重组HMGB1蛋白组(IMP-H+rHMGB1组,25 mmol/L葡萄糖+80μmol/L IMP+200μg/L rHMGB1)。CCK-8试剂盒检测hRECs细胞活性;流式细胞仪检测hRECs细胞凋亡的情况;管腔形成实验检测细胞血管生成;酶联免疫吸附法(ELISA)检测细胞IL-6、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、MDA水平和SOD水平;Western blot检测HMGB1-RAGE-NF-κB信号通路相关蛋白表达情况。结果显示,IMP可以促进高糖诱导的hRECs增殖,且呈浓度依赖性,选择浓度为20、40、80μmol/L的IMP进行后续实验;与NG组比较,HG组细胞存活率、SOD活性降低,细胞凋亡率,管腔形成数,IL-6、TNF-α、IL-1β、MDA水平及HMGB1、RAGE、NF-κB p65蛋白表达水平升高(P<0.05);与HG组比较,IMP-L组、IMP-M组、IMP-H组细胞存活率、SOD活性升高,细胞凋亡率,管腔形成数,IL-6、TNF-α、IL-1β、MDA水平及HMGB1、RAGE、NF-κB p65蛋白表达水平降低,且呈浓度依赖性(P<0.05);与IMP-H组比较,IMP-H+rHMGB1组细胞存活率、SOD活性显著降低,细胞凋亡率,管腔形成数,IL-6、TNF-α、IL-1β、MDA水平及HMGB1、RAGE、NF-κB p65蛋白表达水平显著升高(P<0.05)。结果表明,IMP可能通过抑制HMGB1-RAGENF-κB信号通路,降低炎症反应和细胞凋亡水平,进而减轻高糖诱导的视网膜血管内皮细胞损伤。This study investigated the effect of IMP(imperatorin) on high glucose induced retinal endothelial cell injury by regulating the HMGB1(high-mobility group protein box 1)-RAGE(receptor for advanced glycation end products)-NF-κB(nuclear factor kappa-B) signaling pathway.High glucose induced hRECs in retinal endothelial cells were treated with IMP at a concentration of 10-320 μmol/L to screen for the optimal drug concentration.hRECs(human retinal endothelial cells) were separated into normal glucose group(NG group,5.5 mmol/L glucose),high glucose group(HG group,25 mmol/L glucose),low concentration imperatorin group(IMP-L group,25 mmol/L glucose+20 μmol/L IMP),medium concentration imperatorin group(IMP-M group,25 mmol/L glucose+40 μmol/L IMP),high concentration imperatorin group(IMP-H group,25 mmol/L glucose+80 μmol/L IMP),and high concentration imperatorin+recombinant HMGB1 protein group(IMP-H+rHMGB1 group,25 mmol/L glucose+80 μmol/L IMP+200 μg/L rHMGB1).CCK-8assay kit was applied to detect the activity of hRECs cells.Flow cytometry was applied to detect the apoptosis of hRECs cells.The cell angiogenesis was detected by the lumen formation test.ELISA(enzyme linked immunosorbent assay) was applied to detect levels of IL-6,TNF-α(tumor necrosis factor-α),IL-1β(interleukin-1β),MDA,and SOD in cells.Western blot was applied to detect the expression of proteins related to the HMGB1-RAGE-NF-κB signaling pathway.The results showed that IMP was able to promote the proliferation of high glucose induced hRECs in a concentration dependent manner,subsequently,IMP concentrations of 20,40,and 80 μmol/L were selected for subsequent experiments.Compared with the NG group,the cell survival rate and SOD activity in the HG group were decreased;the apoptosis rate,the number of lumen formation,the levels of IL-6,TNF-α,IL-1β,MDA,and the expression of HMGB1,RAGE,NF-κB p65 proteins were increased(P<0.05).Compared with the HG group,the cell survival rate and SOD activity were increased in the IMP-L group,IMP-M group,and IMP-H

关 键 词：欧前胡素 高迁移率族蛋白B1-晚期糖基化终产物受体–核转录因子κB 高糖 视网膜内皮细胞 损伤 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]