摘要
目的探讨D-阿洛糖(D-allose)对小鼠脑缺血再灌注损伤(CIRI)的神经功能恢复、半乳糖凝集素-3(Gal-3)、腺苷一磷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白(AMPK/mTOR)及部分炎症因子表达的影响。方法C57BL/6雄性小鼠50只,随机分为对照组(Con组)、假手术组(Sham组)、脑缺血再灌注损伤组(MCAO组)、脑缺血再灌注损伤+D-阿洛糖组(MCAO+D-allose组)及脑缺血再灌注损伤+改良柑橘果胶组(MCAO+MCP组)。采用线栓法建立大脑中动脉闭塞/再灌注(MCAO/R)模型。造模成功后对小鼠进行Longa神经功能评分及转棒行走评分;采用TTC染色法观察脑梗死灶体积;通过Western blot及RT-PCR技术检测Gal-3、自噬相关分子的表达水平;免疫荧光技术检测脑组织中Gal-3的分布情况;试剂盒检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-8分泌。结果相较于Con组和Sham组,MCAO模型可增加小鼠神经功能评分(P<0.01)、增加脑梗死体积(P<0.01)、上调Gal-3的表达并增强自噬(P<0.01)。给予D-阿洛糖及Gal-3抑制剂MCP治疗后可改善神经功能障碍、减小脑梗死体积(P<0.01),并降低Gal-3的表达(P<0.01),抑制AMPK磷酸化,促进mTOR磷酸化,抑制自噬(P<0.01)。结论D-阿洛糖可有效促进CIRI小鼠神经功能恢复和减小梗死灶体积,其机制可能是通过下调Gal-3的表达进而抑制组织损伤后细胞过度自噬,并减少TNF-α,IL-8等炎性因子释放,从而发挥神经保护作用。
Objective To investigate the effects of D-allose on the restoration of neurological function,Galectin-3(Gal-3),adenosine monophosphate-activated protein kinase/mammalian target of rapamycin(AMPK/mTOR)and the expression of some inflammatory factors in ischemia-reperfusion injury(CIRI)mice.Methods A total of 50 male mice were randomly divided into control group(Con group),sham group(Sham group),cerebral ischemia-reperfusion injury group(MCAO group),cerebral ischemia-reperfusion injury+D-alolose group(MCAO+D-allose group)and cerebral ischemia-reperfusion injury+modified citrus pectin group(MCAO+MCP group).The middle cerebral artery occlusion/reperfusion(MCAO/R)model(reperfusion after 2 hours of MCA ischemia)was established by thread embolism.After successful modeling,the neurological function of mice was evaluated Longa score and rotated rod walking.TTC staining was used to observe the volume of cerebral infarction foci.The expression levels of Gal-3 and autophagy-related molecules were detected by Western blot and RT-PCR.Immunofluorescence was applied to detect the distribution of Gal-3 in brain tissue,and TNF-α,IL-8 secretion was detected with ELISA KIT.Results Compared with Con group and Sham group,the MCAO model represented significant increase in the Longa neurofunction score(P<0.01),cerebral infarction volume(P<0.01),Gal-3 expression and manifasted enhanced autophagy(P<0.01).After treatment with D-allose,it could significantly improve neurological dysfunction,reduce cerebral infarction volume(P<0.01),reduce the expression of Gal-3(P<0.01),inhibit AMPK phosphorylation,promote mTOR phosphorylation,and inhibit autophagy(P<0.01).The use of the Gal-3 inhibitor MCP alone could also achieve the effect of inhibiting autophagy.Conclusion D-allose can effectively promote the recovery of neurological function and reduce the volume of infarct foci in CIRI mice.The mechanism may involve inhibiting excessive cell autophagy by downregulating the expression of Gal-3,and reducing the release of inflammatory factors such as TNF-αand IL-8,thereby exerting neuroprotective effects.
作者
罗耀文
程俊凯
张敏
苟茂荣
李娟
张磊
高大宽
Luo Yaowen;Cheng Junkai;Zhang Min;Gou Maorong;Li Juan;Zhang Lei;Gao Dakuan(Postgraduate Office,Xi′an Medical University,Xi′an 710068;Dept of Neurosurgery,Xijing Hospital,Air Force Medical University of PLA,Xi′an 710032)
出处
《安徽医科大学学报》
CAS
北大核心
2023年第9期1467-1473,共7页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81971227)
陕西省自然科学基础研究计划项目(编号:2019KW-039)。
