摘要
目的:观察电针“风府”“太冲”“足三里”穴对帕金森病(PD)小鼠脑黑质酪氨酸羟化酶(TH)、结肠核转录因子-κB(NF-κB)、白细胞介素-6(IL-6)、紧密连接蛋白-1(ZO-1)的影响,探讨电针治疗PD的可能机制。方法:C57BL/6小鼠随机分为对照组、模型组、电针组,每组10只。采用鱼藤酮连续灌胃4周复制PD模型。电针组电针“风府”“太冲”“足三里”穴,30 min/次,1次/d,连续治疗2周。观察各组小鼠治疗前后的行为学改变并评分;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TH表达量;HE染色法观察结肠组织结构变化;实时荧光定量PCR法检测结肠NF-κB、IL-6、ZO-1 mRNA的表达水平;蛋白免疫印迹法检测结肠NF-κB蛋白表达水平;ELISA法检测结肠IL-6含量。结果:与对照组比较,治疗前模型组、电针组以及治疗后模型组小鼠行为学评分升高(P<0.01);与模型组比较,治疗后电针组行为学评分降低(P<0.01)。与对照组比较,模型组小鼠结肠组织杯状细胞、隐窝减少,肌层变薄;与模型组比较,电针组小鼠结肠组织表面绒毛较完整,杯状细胞、隐窝增加,肌层增厚。与对照组比较,模型组旷场实验自主运动总路程、中脑黑质TH表达、结肠ZO-1 mRNA表达水平均显著降低(P<0.01),结肠NF-κB、IL-6蛋白和mRNA表达水平及p-NF-κB/NF-κB比值均显著升高(P<0.01);与模型组比较,电针组旷场实验自主运动总路程、中脑黑质TH表达、结肠ZO-1 mRNA表达水平均显著升高(P<0.01),结肠NF-κB、IL-6蛋白和mRNA表达水平及p-NF-κB/NF-κB比值均显著降低(P<0.01,P<0.05)。结论:电针“风府”“太冲”“足三里”穴能够调节NF-κB/IL-6表达水平,抑制肠道炎性反应,修护PD小鼠肠道屏障功能,增强TH活性,从而改善PD小鼠的行为学表现。
Objective To observe the effects of electroacupuncture(EA)at“Fengfu”(GV16),“Taichong”(LR3),and“Zusanli”(ST36)on the tyrosine hydroxylase(TH),nuclear factor-kappa B(NF-κB),interleukin(IL)-6,and zonula occludens-1(ZO-1)in mice with Parkinson’s disease(PD),and to explore the underlying mechanism of EA in the treatment of PD.Methods C57 BL/6 mice were randomly divided into control,model,and EA groups,with 10 mice in each group.The PD model was induced by rotenone(i.g.)in mice for 28 d.EA was applied to GV16,LR3 and ST36 of mice in the EA group for 30 min,once daily for 14 d.The behavioral changes of mice in each group before and after treatment were observed and scored.The total distance traveled autonomously of mice was detected in the open field test.TH expression in the substantia nigra(SN)was measured by immunohistochemistry,and the changes in colon tissue structure were observed by HE staining.The intestinal mRNA expression of ZO-1,NF-κB,and IL-6 was detected by quantitative real-time PCR,and the colon tissue protein expression of NF-κB was detected by Western blot.The colon tissue concentration of IL-6 was detected by ELISA.Results Compared with the control group,the pre-and post-treatment behavioral scores of the model group and the pre-treatment behavioral score of the EA group were increased(P<0.01).Compared with the model group,the EA group showed reduced behavioral score after treatment(P<0.01).Compared with the control group,the model group showed reduced goblet cells and crypts and thinner muscle layer in the intestinal specimens of mice.Compared with the model group,the EA group showed intact surface villi,increased goblet cells and crypts,and thickened muscle layer.Compared with the control group,the model group exhibited reduced total distance traveled in the open field test,TH expression in SN,and ZO-1 mRNA expression(P<0.01),and elevated mRNA and protein expression of NF-κB and IL-6,and the ratio of p-NF-κB/NF-κB in the intestinal tract(P<0.01).Compared with the model group,the EA group displayed increased total distance traveled,TH expression in SN,and ZO-1 mRNA expression(P<0.01),and declining mRNA and protein expression of NF-κB and IL-6,and the ratio of p-NF-κB/NF-κB in the colon tissue(P<0.01,P<0.05).Conclusion EA at GV16,LR3,and ST36 can regulate the expression of NF-κB/IL-6,inhibit the transmission of the colon tissue inflammatory response,repair the intestinal barrier function,and potentiate the TH activity,thereby improving the behavioral performance of PD mice.
作者
汪瑶
马骏
王彦春
关庆亚
WANG Yao;MA Jun;WANG Yan-chun;GUAN Qing-ya(College of Acupuncture and Orthopedics of Hubei University of Chinese Medicine/Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture-moxibustion,Wuhan 430060,China;College of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430060;Zhongjing Institute of Traditional Chinese Medicine,Henan University of Chinese Medicine,Zhengzhou 450046)
出处
《针刺研究》
CAS
CSCD
北大核心
2022年第5期449-454,共6页
Acupuncture Research
基金
国家重点研发计划项目(No.2020YFC0845800)
国家自然科学基金项目(No.81473788)。
