青蒿琥酯联合胰岛素对伴1型糖尿病牙周病大鼠胰腺组织细胞凋亡的影响  

Effect of artesunate combined with insulin on apoptosis of pancreatic cell in type 1 diabetic rats with periodontal disease

作　　者：姚兆丰 梁晨 农晓琳[1] Yao Zhaofeng;Liang Chen;Nong Xiaolin(College of Stomatology,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学口腔医学院/附属口腔医院,南宁530021

出　　处：《广西医科大学学报》2023年第5期762-769,共8页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81860726);广西医疗卫生适宜技术开发与推广应用项目(No.S2019059)。

摘　　要：目的:探究青蒿琥酯(ART)联合胰岛素(INS)对伴1型糖尿病牙周病大鼠胰腺组织细胞凋亡的影响,并初步探索其机制。方法:50只雄性SD大鼠随机分为5组:正常对照组(NC组)、伴糖尿病牙周病组(DM+PD组)、伴糖尿病牙周病ART干预组(ART组)、伴糖尿病牙周病INS干预组(INS组)、伴糖尿病牙周病ART联合INS干预组(ART+INS组),每组10只;建模成功后每日给药1次,用药4周过后过量麻醉处死,测量各组大鼠空腹血糖值(FBG);酶联免疫吸附测定(ELISA)法检测血清肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)表达水平;微计算机断层扫描技术(Micro-CT)检测牙槽骨吸收情况;苏木精—伊红(HE)染色及免疫组织化学(IHC)法观察胰腺组织病理学改变及Caspase-3、Bax、Bcl-2蛋白在胰岛的表达情况;实时荧光定量聚合酶链式反应(RT-qPCR)和蛋白质免疫印迹(western blotting)法检测胰腺Caspase-3、Bax、Bcl-2 mRNA和蛋白表达水平。结果:与NC组比较,DM+PD组FBG明显升高(P<0.05);血清TNF-α、IL-1β表达水平明显升高(P<0.05);牙槽骨显著吸收(P<0.05);胰岛病理组织学损伤严重;胰腺组织Caspase-3、Bax mRNA及蛋白表达水平升高(P<0.05),Bcl-2 mRNA及蛋白表达水平下降(P<0.05)。与DM+PD组比较,ART组、INS组、ART+INS组大鼠FBG均有所下降(P<0.05);血清TNF-α、IL-1β表达水平显著降低(P<0.05);牙槽骨吸收量减少(P<0.05);胰岛病理组织学损伤有所改善;胰腺组织Caspase-3、Bax mRNA及蛋白表达水平下降(P<0.05),Bcl-2 mRNA及蛋白表达水平上升(P<0.05);其中ART+INS组效果优于ART组和INS组(P<0.05)。结论:ART对伴1型糖尿病牙周病大鼠胰腺组织具有保护作用,与INS联合用药效果更佳,其机制可能与抑制炎症和调控Caspase-3/Bax/Bcl-2凋亡信号通路有关。Objective:To investigate the effect of artesunate(ART)combined with insulin(INS)on apoptosis of pancreatic cell in type 1 diabetic rats with periodontal disease and preliminarily explore its mechanism.Methods:A total of 50 male SD rats were randomly divided into 5 groups:normal control group(NC group),diabetes mellitus with periodontal disease group(DM+PD group),ART intervention group with DM and PD(ART group),INS intervention group with DM and PD(INS group),and ART+INS intervention group with DM and PD(ART+INS group),with 10 rats in each group.Following successful modeling,the rats were administered the drug once a day and after four weeks,they were sacrificed under overdose anesthesia.Fasting blood glucose levels(FBG)were measured in all groups of rats.The serum levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA).Alveolar bone resorption was detected by micro-computed tomography(Micro-CT).The histopathological changes of the pancreas and the expressions of Caspase-3,Bax and Bcl-2 proteins in islets were observed by hematoxylin-eosin(HE)staining and immunohistochemistry(IHC).The mRNA and protein expression levels of Caspase-3,Bax and Bcl-2 in pancreatic tissue were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and western blotting.Results:Compared with NC group,the FBG of rats in DM+PD group significantly increased(P<0.05);the serum levels of TNF-αand IL-1βsignificantly increased(P<0.05);the alveolar bone resorption was significant(P<0.05);the histopathological damage of pancreatic islets was severe;the mRNA and protein expression levels of Caspase-3 and Bax in pancreatic tissue increased(P<0.05),while the mRNA and protein expression levels of Bcl-2 decreased(P<0.05).Compared with DM+PD group,the FBG of rats in ART group,INS group and ART+INS group decreased(P<0.05);the serum levels of TNF-αand IL-1βsignificantly decreased(P<0.05);the alveolar bone resorption decreased(P<0.05);the histopathological

关 键 词：1型糖尿病 牙周病 青蒿琥酯 胰腺 凋亡 

分 类 号：R587.1[医药卫生—内分泌] R781.4[医药卫生—内科学]