Objectives: To observe the effects of electroacupuncture (EA) at "Fengfu" (GV16), "Taichong" (LR3) and "Zusanli" (ST36) on α-synuclein (α-syn), Occludin, Claudin-1, thioredoxin interaction protein (TXNIP) and Nod-like receptor 3 (NLRP3) in Parkinson's disease (PD) mice, so as to investigate the mechanisms of EA on intestinal barrier function and inflammation in PD mice.
Methods: Thirty six C57BL/6 mice were randomly divided into control, model and EA groups, with 12 mice in each group. PD mice model was induced by rotenone intragastric administration for 28 days. Mice in the EA group were treated with EA (2 Hz, 1 mA) at GV16, LR3 and ST36 for 30 min, once a day for 14 days. The behavioral scores were observed. The total distance of autonomic movement was measured by open field test. The expression level of α-syn in substantia nigra and colon tissue was determined by immunohistochemistry. The colonic morphology and goblet cell distribution were observed by Alcian blue staining. The expression levels of Occludin, Claudin-1, TXNIP and NLRP3 mRNA in colon tissue were detected by real-time fluorescence quantitative PCR.
Results: Compared with the control group, the behavioral scores of rats were increased (P<0.01)；the total distance of autonomous movement was decreased (P<0.01)；the positive expression level of α-syn in the substantia nigra and colon was increased (P<0.01)；the goblet cells and crypts in colon tissue were reduced, and the muscular layer was thinner；the expression levels of Occludin and Claudin-1 mRNAs in colon tissue were decreased (P<0.01) while TXNIP and NLRP3 mRNAs were increased (P<0.01) in the model group. Compared with the model group, the surface villi of colon tissue was more complete, the goblet cells and crypts were increased, and the muscular layer was thickened；the other indexes were reversed (P<0.01, P<0.05) in the EA group.
Conclusions: EA at GV16, LR3 and ST36 can reduce the abnormal accumulation of α-syn in the substania nigra and colon tissue of PD mice, alleviate the damage of intestinal barrier, regulate TXNIP/NLRP3 signaling pathway, so as to delay the occurrence and development of PD.
目的: 观察电针“风府”“太冲”“足三里”对帕金森病（PD）小鼠α-突触核蛋白（α-syn）、咬合蛋白（Occludin）、闭合蛋白-1（Claudin-1）及硫氧还蛋白互作蛋白（TXNIP）、NOD样受体3（NLRP3）的影响，探讨电针对PD肠道屏障功能及炎性反应的影响及作用机制。方法: C57BL/6小鼠随机分为对照组、模型组、电针组，每组12只。采用鱼藤酮灌胃复制PD小鼠模型。电针组予“风府”“太冲”“足三里”电针治疗，30 min/次，1次/d，连续治疗14 d。观察各组小鼠行为学评分，旷场实验检测小鼠自主运动总路程，免疫组织化学法检测小鼠中脑黑质区和结肠组织中α-syn的阳性表达，阿尔新蓝染色法观察结肠形态及杯状细胞分布情况，实时荧光定量PCR法检测结肠组织中Occludin、Claudin-1、TXNIP、NLRP3 mRNA表达水平。结果: 与对照组比较，模型组大鼠行为学评分升高（P<0.01）；旷场实验自主运动总路程减少（P<0.01）；中脑黑质区和结肠组织α-syn阳性表达升高（P<0.01）；结肠组织中杯状细胞、隐窝减少，肌层变薄；结肠组织中Occludin、Claudin-1 mRNA表达水平降低（P<0.01），TXNIP、NLRP3 mRNA表达水平升高（P<0.01）。与模型组比较，电针组大鼠行为学评分降低（P<0.01）；旷场实验自主运动总路程增加（P<0.01）；中脑黑质区和结肠组织α-syn阳性表达降低（P<0.01）；结肠组织中表面绒毛较完整，杯状细胞、隐窝增加，肌层增厚；结肠组织中Occludin、Claudin-1 mRNA表达水平升高（P<0.05），TXNIP、NLRP3 mRNA表达水平降低（P<0.01）。结论: 电针“风府”“太冲”“足三里”可清除PD小鼠中脑黑质区和结肠组织中异常聚集的α-syn，改善肠道屏障受损，调节TXNIP/NLRP3信号通路，延缓PD的发生发展。.
Keywords: Electroacupuncture; Intestinal barrier; Nod-like receptor 3; Parkinson’s disease; Thioredoxin interaction protein; α-synuclein.