帕金森病食蟹猴模型回肠结构与功能相关蛋白变化的研究  

Structural and functional changes in the ileum of Parkinsonian cynomolgus monkeys

作　　者：姜雪 张瑜玲 黄鑫鑫 刘斌[2] 李伟鹏 岳峰 Jiang Xue;Zhang Yuling;Huang Xinxin;Liu Bin;Li Weipeng;Yue Feng(School of Medical,Guangxi University,Nanning 530004,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西大学医学院,南宁530004 [2]海南省生物医学工程重点实验室海南大学生物医学工程学院

出　　处：《中华老年心脑血管病杂志》2023年第12期1374-1377,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家重点研发计划项目(2018YFA0108500);海南省重点研发计划课题(ZDYF2021SHFZ049);海南省自然科学基金高层次人才项目(322RC588)。

摘　　要：目的探究帕金森病食蟹猴模型研究其肠道结构与功能的变化,为帕金森病患者出现便秘等肠道症状的机制及其干预提供科学依据。方法选取6只雄性食蟹猴,其中3只使用颈内动脉注射1-甲基-4-苯基-1,2,3,6-四氢吡啶的方式进行造模并出现帕金森病运动症状的食蟹猴,根据Kurlan评分>10分且持续3个月未出现消退迹象为帕金森病组,另外3只未经干预的食蟹猴为对照组。收集2组脑组织与回肠组织,用免疫组织化学染色法酪氨酸羟化酶(TH)分析脑内黑质区多巴胺能神经元数量以及肠道外周TH变化,用苏木精-伊红染色分析肠道的结构,并用闭锁小带蛋白1(ZO-1)评估肠道屏障功能,肠道起搏细胞标志物原癌基因(c-kit)、蛋白基因产物9.5(PGP 9.5)评估肠道蠕动功能。结果帕金森病组左、右两侧黑质多巴胺能神经元数目明显低于对照组[(133.13±108.63)个vs(957.21±225.56)个,t=5.705,P=0.005;(155.74±62.48)个vs(917.52±161.14)个,t=7.611,P=0.002],对照组回肠肌间神经丛TH吸光度值明显高于帕金森病组(0.79±0.01vs0.73±0.01,t=5.149,P=0.007),帕金森病组绒毛密度低于对照组,且排布不规律、结构散乱、细胞脱落,对照组回肠黏膜ZO-1吸光度值(0.28±0.01vs0.22±0.02,P=0.006),肌间神经丛c-kit吸光度值(0.21±0.01vs0.18±0.01,P=0.007)明显高于帕金森病组,对照组与帕金森病组肌间神经丛PGP9.5表达比较,差异无统计学意义,(0.31±0.08vs0.38±0.06,P=0.322)。结论帕金森病组肠道屏障完整性的破坏,包括绒毛受损、肠道通透性增加以及蠕动性降低,与蠕动性功能相关的肌间神经元无关。Objective To study the changes in intestinal structure and function of Parkinson’s dis-ease(PD)model in cynomolgus monkeys so as to provide scientific basis for exploring the mecha-nism and intervention of constipation and other intestinal symptoms in PD patients.Methods Six maee cynomolgus monkeys were subjected,and three of them received intracarotid injection of MPTP for manifestations of typical PD motor symptoms.The monkeys with the Kurlan score reached 10 and persisting for over 3 months without abrogation were identified as PD model(PD group).The other three monkeys served as control group.The brain and ileum tissue samples were collected from the two groups of monkeys.Immunohistochemical staining with tyrosine hydroxy-lase(TH)was used to determine the number of dopaminergic neurons in the substantia nigra of the brain and the changes in peripheral TH of the intestine.HE staining was employed to observe the structure of the intestine.Intestinal permeability was evaluated by atresia occlusive zone pro-tein 1(ZO-1),and intestinal peristalsis function was assessed with intestinal pacemaker cell biomarkers oncogene c-kit and protein gene product 9.5(PGP9.5).Results When compared with the control group,the PD group had significantly less number of nigrostriatal dopaminergic neurons in the left and right sides(133.13±108.63 vs 957.21±225.56,t=5.705,P=0.005;155.74±62.48 vs 917.52±161.14,t=7.611,P=0.002),lower expression of TH in the ileal intermuscular plexus(0.79±0.01 vs 0.73±0.01,t=5.149,P=0.007),reduced villus density with irregular arrange-ment,scattered structure,and detachment of goblet cells,and increased expression of ZO-1(0.28±0.01 vs 0.22±0.02,P=0.006)and c-kit(0.21±0.01 vs 0.18±0.01,P=0.007)in the ileum muco-sa,but there was no such difference in the expression of PGP9.5 between the two groups(0.31±0.08 vs 0.38±0.06,P=0.322).Conclusion Intestinal barrier integrity is damaged in PD mon-keys,with damaged villi,increased intestinal permeability and decreased peristalsis,and peristalticfuncti

关 键 词：帕金森病 成束猴 模型 动物 便秘 回肠 闭锁小带蛋白-1 

分 类 号：R-332[医药卫生] R742.5