电针对鱼藤酮诱导的帕金森病模型大鼠黑质内c-Jun氨基末端激酶及γ干扰素的影响

Effect of electroacupuncture on c-Jun amino terminal kinase signaling pathway and inflammatory cytokines interferon-γ in substantia nigra cells of rotenone-induced rats model of Parkinson＇s disease

目的 观察电针对帕金森病（PD）模型大鼠黑质内c-Jun氨基末端激酶（JNK）信号通路及γ-干扰素（IFN-γ）的影响.方法 选取雄性健康SD大鼠32只,按照随机数字表法将其分为正常组、假手术组、模型组及电针组,每组8只.模型组和电针组大鼠经颈背部注射鱼藤酮[1 mg/kg,溶于一定比例二甲亚砜（DMSO）和生理盐水中,浓度0.25 mg/ml],假手术组经颈背部注射同剂量的DMSO和生理盐水混合液,正常组不进行特殊干预.PD大鼠造模成功后,在电针组大鼠“风府”、“太冲”穴位进行治疗.治疗结束后,对4组大鼠进行行为学观察及敞箱实验.采用免疫蛋白印迹法检测4组大鼠脑内酪氨酸羟化酶（TH）、磷酸化c-Jun和IFN-γ的表达情况.结果 模型组大鼠表现出明显的PD综合征,与正常组和假手术组比较,差异无统计学意义（P＞0.05）.敞箱实验结果显示,模型组大鼠水平运动[（19.12±2.34）分]、垂直运动活性[（5.27±1.04）分]较正常组和假手术组明显降低,差异有统计学意义（P＜0.05）.电针治疗后,大鼠运动活性较模型组明显增强,差异有统计学意义（P＜0.05）,与正常组和假手术组比较,差异无统计学意义（P＞0.05）.与正常组比较,模型组大鼠黑质区TH蛋白（0.183 ±0.021）表达显著减少,磷酸化c-Jun（0.388±0.028）和IFN-γ蛋白（0.453 ±0.033）表达显著增加,差异有统计学意义（P＜0.05）.与正常组比较,电针组大鼠黑质区TH蛋白（0.324 ±0.054）表达有所减少,磷酸化的c-Jun（0.207±0.059）和IFN-γ蛋白（0.239±0.022）表达有所增加,差异无统计学意义（P＞0.05）.与模型组比较,电针组大鼠黑质区TH蛋白明显增加,磷酸化的c-Jun和IFN-γ蛋白表达明显减少,差异有统计学意义（P＜0.05）.结论 电针刺激可降低大鼠黑质区c-Jun及炎症因子IFN-γ的表达水平,对PD模型大鼠体内JNK信号通路及疾病进展起到一定的调节作用.

Objective To observe the effects of electroacupuncture （EA） on c-Jun amino terminal kinase （JNK） signaling pathway and inflammatory cytokines interferon-γ （IFN-γ） in substantia nigra （SN） cells of rotenone-induced rats model of Parkinson＇s disease （PD）,and explore the underlying mechanism of EA on PD.Methods A total of 32 male Sprague-Dawley mice were randomly and evenly divided into a normal group,a shamoperation group,a model group and an EA group.Model group and EA group were injected intradermally with rotenone （1 mg/kg,dissolved in DMSO and saline,concentration：O.25 mg/ml） on the nape of neck.Sham-operation group was injected the same dose of DMSO and saline.Normal group had no intervention.EA group was applied to Fengfu （DU16） and Taichong （LR3） acupoints after the establishment of PD model in rats.Behavioral assessment was conducted after the treatments,the rats were sacrificed for sampling substantia nigra tissue to detect the expressions of tyrosine hydroxylase （TH）,p-c-Jun amino terminal kinase （p-c-Jun） and interferon-γ（IFN-γ） protein with Western blotting （WB）.Results Model rats showed significant PD syndrome characteristics,comparing with normal group and sham group,the difference was not statistically significant （P ＞ 0.05）.The results of open box test showed that the scores of model group rats decreased significantly in terms of the horizontal movement [（19.12 ±2.34） points] and vertical locomotor activity [（5.27 ± 1.04） points] when compared with normal group and sham group,the difference was statistically significant （P ＜ 0.05）.After EA treatment,locomotor activity of rats increased significantly when compared with model group （P ＜ 0.05）,however,the normal group and sham group was not statistically and significantly different in locomotor activity （P ＞ 0.05）.Compared with normal group,the expression of TH protein in （0.183 ± 0.0213） reduced significantly and the expressions of p-c-Jun （0.388 ± 0.0283） and IFN-γ protein（0.453 ± 0.0332） increased significantly in model group,the difference was statistically significant （P ＜0.05）.Compared with normal group,the expression of TH protein（0.324 ± 0.0538） reduced and the expressions of p-c-Jun（0.207 ± 0.0592） and IFN-γ protein （0.239 ± 0.0215） increased in EA group,the difference was not statistically significant （P ＞ 0.05）.Compared with model group,the expression of TH protein increased significantly in EA group（P ＜ 0.05）,the expressions of p-c-Jun and IFN-γ protein reduced significantly in EA group（P ＜ 0.05）.Conclusion EA therapy may reduce the expression of IFN-γ protein in SN of PD rats model by regulating the expression of JNK/mitogen-activated protein kinase （MAPK） pathway,which may delay the process of PD.