贝那鲁肽对高糖诱导的胰岛β细胞功能障碍和凋亡的影响及机制研究  

Effect and Mechanism of Benaglutide on IsletβCell Dysfunction and Apoptosis Induced by High Glucose

作　　者：辛丽娟 宁建峰 唐宏霞 左源渊 王新婷 彭一 尚宏博 鲍喜静 XIN Li-juan;NING Jian-feng;TANG Hong-xia;ZUO Yuan-yuan;WANG Xin-ting;PENG Yi;SHANG Hong-bo;BAO Xi-jing(Department of EndocrinologyⅡ,the First Hospital of Zhangjiakou,Hebei Zhangjiakou 075000,China;Department of Clinical Laboratory,the First Hospital of Zhangjiakou,Hebei Zhangjiakou 075000,China;Endocrine and Metabolic Diseases Clinic,the First Hospital of Zhangjiakou,Hebei Zhangjiakou 075000,China;Department of EndocrinologyⅠ,the First Hospital of Zhangjiakou,Hebei Zhangjiakou 075000,China)

机构地区：[1]张家口市第一医院内分泌二科,河北张家口075000 [2]张家口市第一医院检验科,河北张家口075000 [3]张家口市第一医院内分泌代谢病门诊,河北张家口075000 [4]张家口市第一医院内分泌一科,河北张家口075000

出　　处：《现代检验医学杂志》2022年第1期103-106,113,共5页Journal of Modern Laboratory Medicine

基　　金：2019年度张家口市科技计划项目(编号:1921081D)。

摘　　要：目的探讨贝那鲁肽对高糖诱导的胰岛β细胞功能障碍和凋亡的影响以及相关机制。方法将INS-1胰岛β细胞随机分为对照组、高糖组、贝那鲁肽处理组、MG-132预处理组和ISO-1预处理组,对照组给予5.5mmol/L葡萄糖处理,高糖组给予30 mmol/L葡萄糖处理,贝那鲁肽处理组在高糖组的基础上给予1 nmol/L贝那鲁肽处理,MG-132预处理组和ISO-1预处理组分别在高糖的基础上给予20μmol/L MG-132和50μmol/L ISO-1预处理。培养24h后噻唑蓝(MTT)比色法评估细胞的活力,酶联免疫吸附法(ELISA)检测INS-1胰岛β细胞胰岛素和巨噬细胞迁移抑制因子(MIF)的分泌,Western blot检测凋亡相关蛋白(Bax和cleaved caspase 3)和NF-κB通路相关蛋白(p-IκB,IκB和NF-κBp65)的表达。结果与对照组相比,高糖组胰岛β细胞的存活率(t=4.949,P<0.01)、胰岛素的分泌(t=3.168,4.273,均P<0.05)和IκB的表达(t=3.062,P<0.05)明显降低,凋亡相关蛋白(Bax和cleaved caspase 3)的表达(t=2.923,3.141,均P<0.05)、p-IκB和NF-κB p65表达(t=3.544,4.658,均P<0.01)以及MIF分泌(t=3.024,P<0.05)明显增加.与高糖组相比,贝那鲁肽处理组可逆转高糖组上述指标的变化(t=2.415~4.290,均P<0.05),差异均有统计学意义。进一步给予NF-κB抑制剂MG-132和MIF抑制剂ISO-1预处理后,与高糖组相比,MG-132预处理组和ISO-1预处理组均可改善高糖诱导的INS-1胰岛β细胞胰岛素分泌功能(t=2.515~6.867,均P<0.05),还可降低凋亡相关蛋白Bax(t=4.022,2.554,均P<0.05)和cleaved caspase 3(t=4.022,2.760,均P<0.05)的表达,差异均有统计学意义。结论贝那鲁肽可通过抑制NF-κB/MIF依赖的炎症途径减轻高糖诱导的胰岛β细胞功能障碍和凋亡。Objective To investigate the effects and mechanisms of benaglutide onhigh glucose-induced isletβcell dysfunction and apoptosis.Methods INS-1 isletβcells were randomly divided into control group,high glucose group,benaglutide treatment group,MG-132 pretreatment group and ISO-1 pretreatment group.The control group was administered 5.5 mmol/L glucose,the high glucose group was administered 30 mmol/L glucose,and the benaglutide treatment group was administered 1 nmol/L benaglutide on the basis of the high glucose group,the MG-132 pretreatment group and ISO-1 pretreatment group were administered 20μmol/L MG-132 and 50μmol/L ISO-1 on the basis of high glucose group.The cells viability,the levels of insulin and macrophage migration inhibitory factor(MIF),and the expression of apoptosis-related proteins(Bax and cleared caspase 3)and NF-κB pathway-related proteins(p-IκB,IκB and NF-κB p65)in INS-1 islet β cells were respectively evaluated by the MTT assay,enzyme-linked immunosorbent assay(ELISA),and Western blot after 24 hours of culture.Results Compared with the control group,the cells viability(t=4.949,P<0.01),insulin secretion(t=3.168,4.273,all P<0.05),and IκB expression(t=3.062,P<0.05)of islet β cells in the high glucose group were significantly reduced,apoptosis-related proteins(Bax and cleared caspase 3)(t=2.923,3.141,all P<0.05),p-IκB(t=3.544,P<0.01)and NF-κB p65(t=4.658,P<0.01)expression and MIF secretion(t=3.024,P<0.05)were significantly increased.Compared with the high glucose group,the changes of above indicators were reversed in benaglutide treatment group(t=2.415~4.290,all P<0.05),the differences were statistically significant respectively.Furthermore,compared with the high glucose group,the MG-132 pretreatment group and the ISO-1 pretreatment group couldimprove the high glucose-induced INS-1 Insulin β cells insulin secretion function(t=2.515~6.867,all P<0.05),andreduce apoptosis-related proteins Bax(t=4.022,2.554,all P<0.05)and cleared caspase 3(t=4.0222.760,all P<0.05)expression,the differences

关 键 词：贝那鲁肽 INS-1胰岛β细胞 NF-ΚB 巨噬细胞迁移抑制因子 凋亡 

分 类 号：R587[医药卫生—内分泌] R446[医药卫生—内科学]