脂多糖对肠道α-突触核蛋白表达的影响  

Effect of lipopolysaccharide on the expression of α-synuclein in the intestinal tract

作　　者：刘梦茹 宫晓丽[2,3] 王乐[1,2] 刘玚[1,2] 张婷 王晓民[1,2] Liu Mengru;Gong Xiaoli;Wang Le;Liu Yang;Zhang Ting;Wang Xiaomin(Department of Neurobiology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Key Laboratory of Neurodegenerative Disorders of the Ministry of Education,Capital Medical University,Beijing 100069,China;Department of Physiology and Pathophysiology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学基础医学院神经生物学系,北京100069 [2]首都医科大学教育部神经变性病重点实验室,北京100069 [3]首都医科大学基础医学院生理与病理生理学系,北京100069

出　　处：《首都医科大学学报》2018年第4期557-562,共6页Journal of Capital Medical University

基　　金：国家重点研究和发展计划(2016YFC1306300);国家自然科学基金重大项目资助(81527901);北京市自然科学基金(7162019);北京市科学技术委员会国家重大研发计划(Z161100002616007)~~

摘　　要：目的腹腔注射细菌脂多糖(lipopolysaccharide,LPS)构建炎性反应诱导的帕金森病(Parkinson's disease,PD)小鼠模型,检测PD模型小鼠的结肠中α-突触核蛋白(α-synuclein,α-syn)的表达情况,以及结肠组织的炎性反应水平,探索炎性反应对PD早期外周结肠组织中关键致病蛋白α-syn表达的影响。方法将雄性的C57BL/6J小鼠采用数字表法随机分为0.9%(质量分数)氯化钠注射液对照组和模型组,给予小鼠单次腹腔注射5 mg/kg LPS或等体积0.9%(质量分数)氯化钠注射液,通过检测外周血中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的含量和小鼠体质量来评价模型建立情况;通过蛋白免疫印迹检测结肠组织α-突触核蛋白(α-synuclein,α-syn)的表达和炎性反应小体的激活情况;通过反转录-聚合酶链反应(reverse transcriptionpolymerase chain reaction,RT-qPCR)检测结肠组织炎性反应因子TNF-α的转录水平。结果 LPS注射后急性期(3 h内),小鼠结肠α-syn蛋白表达增加,并伴随炎性反应小体NOD样受体家族蛋白3(NOD-like receptor protein 3,NLRP3)蛋白和TNF-αmRNA的表达增加;LPS注射后慢性期(1个月),小鼠结肠α-syn及NLRP3蛋白表达恢复正常水平,但结肠的炎性反应因子TNF-α较对照组仍明显增加。结论 LPS可以促进α-syn在结肠内的表达,并伴随着结肠组织炎性反应小体激活和炎性反应因子TNF-α的表达,一个月时α-syn和炎性反应小体NLRP3恢复至正常水平,但结肠炎性反应因子TNF-α的持续高表达,很可能是导致LPS诱导的PD模型慢性反应期α-syn蛋白再一次持续升高的原因。Objective Intraperitoneal injection of lipopolysaccharide（LPS） was used to construct Parkinson＇s disease（PD） mouse model. The protein level of α-synuclein（α-syn） and intestinal inflammation were detected in intestinal tissue of PD mice,to explore the expression of key pathogenic protein in the intestinal tract of early PD. Methods Male C57 BL/6 J mice were divided into control group and model group,mice received intraperitoneal injection of LPS or 0. 9% saline. The mice model was characterized by detecting the level of inflammatory factor tumor necrosis factor-α（TNF-α） in blood and body weight. The protein level of α-synuclein（α-syn） and inflammasome NOD-like receptor protein 3（NLRP3） in intestine were detected by Western blotting,the expression of TNF-α was measured by reverse transcription-polymerase chain reaction（RT-q PCR）. Results In the acute phase,the expression of α-syn,NLRP3 and TNF-α were increased after LPS injection,the α-syn and NLRP3 returned to baseline levels at one month after LPS administration.But the expression of TNF-α was still at a high level. Conclusion LPS promoted the expression of α-syn in the intestine,which was accompanied by inflammasome activation and TNF-α expression. However,the high level of TNF-α is likely to lead to the progressive increase of α-syn in the chronic period of PD model induced by LPS.

关 键 词：Α-突触核蛋白 炎性反应 肠道 帕金森病 

分 类 号：R741.02[医药卫生—神经病学与精神病学]