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

  • 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。

     

    Abstract: 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 melli-tus 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 ob-served by hematoxylin-eosin (HE) staining and im-munohistochemistry (IHC).The mRNA and protein expression levels of Caspase-3, Bax and Bcl-2 in pancreatic tissue were detected by real-time fluorescence quanti-tative 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-α and IL-1β significantly in-creased (P< 0.05); the alveolar bone resorption was significant (P< 0.05); the histopathological damage of pan-creatic islets was severe; the mRNA and protein expression levels of Caspase-3 and Bax in pancreatic tissue in-creased (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-α and IL-1β significantly decreased (P< 0.05); the alveolar bone resorption decreased (P< 0.05); the histopathological damage of pancreatic islets were alleviated; the mRNA and protein expression levels of Cas-pase-3 and Bax in pancreatic tissue decreased (P< 0.05), while the mRNA and protein expression level of Bcl-2 increased (P< 0.05); the ART+INS group had a better effect than the ART and INS groups (P< 0.05).Conclusion:ART has a protective effect on pancreatic tissue in type 1 diabetic rats with periodontal disease, and the ef-fect of ART combined with INS is better.The mechanism may be related to the inhibition of inflammation and the regulation of Caspase-3/Bax/Bcl-2 apoptosis signaling pathway.

     

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