电针调节NLRP3/Caspase-1通路对帕金森病大鼠多巴胺能神经元焦亡的影响  

Effect of electroacupuncture regulating NLRP3/Caspase-1 pathway on pyroptosis of dopaminergic neurons in rats with Parkinson’s disease

作　　者：刘艳阳 郭雅碧 翟宏宇 雷德宝 王红 赵素晨 刘佩军 LIU Yan-yang;GUO Ya-bi;ZHAI Hong-yu;LEI De-bao;WANG Hong;ZHAO Su-chen;LIU Pei-jun(Department of Rehabilitation Medicine,Xiangyang Central Hospital,the Affiliated Hospital of Hubei University of Arts and Sciences,Xiangyang 441021,HubeiProvince,China)

机构地区：[1]湖北文理学院附属医院襄阳市中心医院康复医学科,湖北襄阳441021

出　　处：《针刺研究》2022年第11期983-992,共10页Acupuncture Research

摘　　要：目的:观察电针对帕金森病(PD)大鼠多巴胺能神经元焦亡的影响,探讨其可能的分子机制。方法:SD大鼠随机分为假手术组、模型组、电针组、MCC950组、电针+MCC950组,每组12只。采用6-羟基多巴胺两点注射法制备PD大鼠模型。MCC950组大鼠按10 mg/kg腹腔注射MCC950,1次/d;电针组予电针“太冲”及“风府”,每次30 min,1次/d;电针+MCC950组在电针组基础上按10 mg/kg给予腹腔注射MCC950,1次/d。以上干预均连续2周。干预后,采用诱导旋转实验、转棒实验和旷场实验观察大鼠行为学改变;免疫组织化学法检测中脑黑质多巴胺能神经元标志物酶酪氨酸羟化酶(TH)、α-突触核蛋白(α-Syn)的阳性表达;HE染色法观察大鼠中脑黑质纹状体多巴胺能神经元损伤程度;免疫荧光双染法检测脑组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、半胱天冬氨酸蛋白酶1(Caspase-1)和离子钙接头蛋白1(Iba-1)的免疫阳性共表达;ELISA法检测脑组织中白细胞介素(IL)-1β、IL-18的含量;Western blot法检测中脑黑质纹状体NLRP3、剪切的半胱天冬氨酸蛋白酶-1(Cleaved Caspase-1)、凋亡相关斑点样蛋白(ASC)的蛋白表达水平。结果:与假手术组比较,模型组大鼠的旋转圈数、中央区域停留时间、中脑黑质α-Syn阳性表达均增加(P<0.05);掉落潜伏期、直立次数、运动总距离、中脑黑质TH阳性表达均减少(P<0.05);脑组织NLRP3、Caspase-1阳性表达,IL-1β、IL-18含量,以及黑质纹状体中NLRP3、ASC、Cleaved Caspase-1蛋白表达水平均升高(P<0.05)。与模型组比较,电针组、MCC950组和电针+MCC950组大鼠的上述指标均发生逆转(P<0.05)。与MCC950组和电针组比较,电针+MCC950组的旋转圈数、中央区域停留时间减少(P<0.05),掉落潜伏期、直立次数和运动总距离增加(P<0.05);中脑黑质TH阳性表达增加(P<0.05),α-Syn阳性表达减少(P<0.05);脑组织NLRP3、Caspase-1阳性表达,IL-1β、IL-18含量,以及黑�Objective To observe the effect of electroacupuncture(EA)on the pyrolysis of dopaminergic neurons in rats with Parkinson’s disease(PD),so as to explore its underlying molecular mechanism.Methods Male SD rats were randomly divided into the sham operation,model,EA,MCC950 and EA+MCC950 groups,with 12 rats in each group.The PD rat model was established by two-point injection of 6-OHDA.Rats in the MCC950 group were injected with MCC950 at the dose of 10 mg/kg,once a day;for rats of EA group,EA was applied to“Taichong”(LR3)and“Fengfu”(GV16)for 30 min,once a day;rats in the EA+MCC950 group were given MCC950 injection and EA once a day.All above interventions were performed for 2 weeks.After the intervention,the behavioral changes of rats were observed using rotating induction experiment,rotating rod experiment and open field experiment;the positive expressions of dopaminergic neuronal markers tyrosine hydroxylase(TH)andα-Synuclein(α-Syn)in substantia nigra striatum were detected by immunohistochemistry;the damage of dopaminergic neurons in substantia nigra striatum was observed after HE staining;immunopositive coexpression of brain nucleotide-binding oligomerization domain like receptor protein 3(NLRP3),Caspase-1 and ionized calcium binding adapter1(Iba-1)were detected by immunofluorescence double staining;the levels of interleukin(IL)-1βand IL-18 in brain tissues were detected by ELISA;the protein expression levels of NLRP3,Cleaved Caspase-1 and apoptosis-associated speck-like protein containing a CARD(ASC)in the substantia nigra striatum were detected by Western blot.Results Compared with the sham operation group,the number of rotations of rotating induction experiment,the residence time in the central area of open field experiment,the positive expression ofα-Syn,the positive co-expressions of NLRP3/Iba-1 and Caspase-1/Iba-1,the contents of IL-1βand IL-18 in brain tissues,and the protein expression levels of NLRP3,ASC and Cleaved Caspase-1 in substantia nigra striatum were significantly increased(P<0.05

关 键 词：帕金森病 电针 神经元焦亡 核苷酸结合寡聚化结构域样受体蛋白3/半胱天冬氨酸蛋白酶-1 

分 类 号：R245.97[医药卫生—针灸推拿学]