电针对帕金森病小鼠肠道NEK7/NLRP3炎症信号通路的影响

Effect of electroacupuncture on the NEK7/NLRP3 inflammatory signaling pathway in intestine of mice with Parkinson′s disease

目的观察电针对帕金森病小鼠肠道NIMA相关激酶7(NEK7)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)信号通路的影响。方法36只C57BL/6小鼠按照随机数字表法分为对照组、模型组、电针组,每组12只。模型组、电针组连续灌胃鱼藤酮溶液(10 mg/kg)28 d建立帕金森病小鼠模型。造模结束后,电针组选取“风府”“太冲”“足三里”电针刺激14 d,对照组、模型组仅固定。爬杆实验、后肢评分检测小鼠运动能力,免疫组织化学法检测小鼠黑质酪氨酸羟化酶(TH)、结肠组织封闭蛋白阳性表达,苏木精-伊红染色法观察结肠组织学形态,蛋白质印迹法检测小鼠结肠组织NEK7、NLRP3、胱天蛋白酶-1(Caspase-1)及白细胞介素-1β(IL-1β)蛋白表达。结果与对照组比较,模型组小鼠爬杆实验评分降低,后肢评分升高(P<0.01);黑质TH、结肠组织封闭蛋白平均光密度降低(P<0.01);结肠组织可见明显炎性浸润,肌层变薄;结肠组织NEK7、NLRP3、Caspase-1、IL-1β蛋白表达升高(P<0.01)。与模型组比较,电针组小鼠爬杆实验评分升高,后肢评分降低(P<0.01);黑质TH、结肠封闭蛋白平均光密度升高(P<0.01);结肠组织炎性浸润程度降低,肌层增厚;结肠组织NEK7、NLRP3、Caspase-1、IL-1β蛋白表达降低(P<0.01)。结论电针“风府”“太冲”“足三里”可改善帕金森病小鼠的运动功能障碍,其机制可能是通过抑制肠道NEK7/NLRP3通路,改善肠屏障损伤,缓解肠道炎症,改善多巴胺能神经元损伤。

Objective To investigate the effects of electroacupuncture on the intestinal NIMA-related kinase 7(NEK7)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway in mice with Parkinson′s disease.Methods According to the randomized number table method,36 C57BL/6 mice were randomly divided into the control group,the model group,and the electroacupuncture group,with 12 mice per group.The Parkinson′s disease mouse model was established by gavage of rotenone solution(10 mg/kg)for 28 d.After molding,the electroacupuncture group was stimulated with"Fengfu"(DU17),"Taichong"(LR3),and"Zusanli"(ST36)for 14 d,while the control group and the model group were only treated with immobilization.The motor ability of mice was detected by pole climbing test and hindlimb rating score,the positive expressions of nigral tyrosine hydroxylase(TH)and colon occludin were detected by immunohistochemistry,the histological morphology of colon was observed by hematoxylin-eosin staining,and Western blotting was used to detect the protein expressions of NEK7,NLRP3,Caspase-1,and interleukin-1β(IL-1β).Results Compared with the control group,mice in the model group had lower score on the pole climbing test and a higher hindlimb rating score(P<0.01);the average optical densities of nigral TH and colon occludin were decreased(P<0.01);significant inflammatory infiltration was observed in the colonic tissue,and the muscularis propria was thinned;and the protein expressions of NEK7,NLRP3,Caspase-1,and IL-1βin the colonic tissue were elevated(P<0.01).Compared with the model group,mice in the electroacupuncture group had higher score on the pole climbing test and a lower hindlimb rating score(P<0.01);the average optical densities of nigral TH and colon occludin were increased(P<0.01);the degree of inflammatory infiltration of colonic tissues decreased,and the muscularis propria was thickened;and the protein expressions of NEK7,NLRP3,Caspase-1,and IL-1βof colonic tissues were decreased(P<0.01).Conclusion Electroacupuncture at"Fengfu"(DU17),"Taichong"(LR3),and"Zusanli"(ST36)can improve motor functional impairments in mice with Parkinson′s disease,and the mechanism may be through the inhibition of intestinal NEK7/NLRP3 pathway,improving the intestinal barrier damage,relieving the intestinal inflammation,and improving the dopaminergic neuron injury.