摘要
目的探讨胰高血糖素样1受体(GLP-1R)的小分子变构激动剂6,7-dichloro-2-methylsulfonyl-3-N-tert-butylaminoquinoxaline(DMB)延迟给药对脑缺血再灌注损伤(CIRI)所致行为缺陷的作用及机制。方法将C57BL/6J成年雄性小鼠随机分为假手术组、模型组和DMB高、中、低剂量组。模型组行大脑中动脉栓塞再灌注(MCAO/R)损伤,DMB高、中、低剂量组在MCAO/R损伤24 h后每日给予不同剂量DMB,连续5 d,假手术组及模型组每日给予等体积生理盐水。以神经功能缺陷评分、足误实验和转棒实验评价小鼠行为学缺陷;检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)和过氧化氢酶(CAT)水平,评价小鼠血液生化指标的变化;以氯化三苯基四氮唑(TTC)染色法检测缺血梗死体积;以TdT介导的dUTP缺口末端标记技术(TUNEL)法检测缺血半暗区神经元细胞凋亡;以免疫荧光法检测缺血半暗区星形胶质细胞标志物神经胶质纤维酸性蛋白(GFAP)表达;用Western Blot及Elisa法检测星形胶质细胞氧糖剥夺/复氧(OGD/R)损伤后白细胞介素-1β(IL^-1β)水平。结果实验表明,DMB延迟给药5 d后能显著改善小鼠行为学缺陷;能降低MCAO/R损伤后血清SOD、MDA和CAT水平;对梗死体积无显著影响;能显著降低缺血半暗区神经元细胞凋亡;能显著降低星形胶质细胞增生及活化水平;能显著降低星形胶质细胞OGD/R损伤后IL^-1β水平。结论DMB延迟给药改善了MCAO/R损伤所致小鼠行为缺陷,其机制可能是通过减轻星形胶质细胞炎性因子释放进而减轻神经元细胞凋亡。
Objective To investigate the effect and mechanism of delayed treatment by a small molecule allosteric agonist 6,7-dichloro-2-methylsulfonyl-3-N-tert-butylaminoquinoxaline(DMB),glucagon-like peptide-1 recipient(GLP-1R),on cerebral ischemia and reperfusion injury(CIRI)induced behavioral deficits.Methods C57BL/6J adult male mice were randomly divided into sham group,model group and DMB high,middle and low dose group.Middle cerebra artery occlusion/reperfusion(MCAO/R)injury was performed in model group.24 h after MCAO/R injury,DMB was administrated in DMB high,middle and low dose group for 5 consecutive days.Sham group and model group were administered the same volume of saline solution every day.Neurological function score,foot fault test and rotarod test were employed to evaluate the behavioral injury of mice;the level of superoxide dismutase(SOD),malondialdehyde(MDA)and catalase(CAT)in serum was detected respectively to examine the changes of blood biochemical indexes in mice;triphenyltetrazolium chloride(TTC)staining method was utilized to detect the volume of ischemic infarction;TdT-mediated dUTP nick end labeling(TUNEL)assay was used to detect neuronal apoptosis in ischemic penumbra;immunofluorescent staining was performed to test the expression of astrocyte marker glial fibrillary acidic protein(GFAP)in ischemic penumbra;Western Blot experiment and Elisa assay were employed to determine interleukin-1β(IL^-1β)level after oxygen glucose deprivation/reperfusion(OGD/R)in astrocytes.Results The results of experiments showed that 5 d after delayed administration of DMB,behavioral deficiency of mice was significantly improved,levels of serum SOD,MDA and CAT after MCAO/R injury were decreased,infarct volume was not obviously influenced,cell apoptosis number in ischemic penumbra was significantly reduced,levels of proliferation and activation in astrocytes were apparently decreased,and IL^-1βlevel after OGD/R injury in astrocytes was significantly downregulated.Conclusion Delayed treatment of DMB could improve the behavioral deficits induced by MCAO/R injury of mice,and the mechanism might be to reduce neuronal apoptosis by reducing the release of astrocyte inflammatory cytokines.
作者
时庭玉
程刘思远
张慧楠
SHI Tingyu;CHENG Liusiyuan;ZHANG Huinan(Department of Basic Medicine,Air Force Military Medical University,Xi′an 710032,China;Department of Pharmacology,School of Pharmacy,Air Force Military Medical University,Xi′an 710032,China)
出处
《西北药学杂志》
CAS
2020年第5期674-679,共6页
Northwest Pharmaceutical Journal
基金
国家自然科学基金项目(编号:81601151)。
