电针对帕金森病大鼠海马组织MAP4K3/MKK4/JNK通路的影响

Effect of electroacupuncture on MAP4K3/MKK4/JNK pathway in hippocampus of rats with Parkinson’s disease

目的探讨电针对帕金森病(PD)大鼠海马组织丝裂原活化蛋白激酶激酶激酶激酶3(MAP4K3)/丝裂原活化蛋白激酶激酶4(MKK4)/c-Jun氨基末端激酶(JNK)通路的影响。方法大鼠分为对照组、PD模型组(PD造模)及电针干预组(PD造模+电针干预)。Morris水迷宫实验记录逃避潜伏期及原平台停留时间,旷场实验统计水平运动得分及垂直运动得分,HE染色观察大鼠海马组织病理改变,TUNEL染色检测大鼠海马组织中细胞凋亡,Western blot法检测大鼠海马组织中MAP4K3/MKK4/JNK通路及凋亡相关蛋白表达。结果与对照组相比,PD模型组大鼠逃避潜伏期显著延长(P<0.05),原平台停留时间显著缩短(P<0.05),水平运动得分及垂直运动得分显著降低(P<0.05),大鼠海马组织中细胞凋亡率及MAP4K3、p-MKK4/MKK4、p-JNK/JNK、Bax、cleaved caspase-3蛋白表达水平显著升高(P<0.05),海马组织出现明显的病理学改变;与PD模型组相比,电针干预组大鼠逃避潜伏期显著缩短(P<0.05),原平台停留时间显著延长(P<0.05),水平运动得分及垂直运动得分显著升高(P<0.05),大鼠海马组织中细胞凋亡率及MAP4K3、p-MKK4/MKK4、p-JNK/JNK、Bax、cleaved caspase-3蛋白表达水平显著降低(P<0.05),海马组织病理改变较轻。结论电针可抑制MAP4K3/MKK4/JNK通路激活,减轻PD大鼠海马组织细胞凋亡及病理改变,提高认知、行为功能。

Objective To investigate the effect of electroacupuncture on the mitogen-activated protein kinase kinase kinase kinase 3(MAP4K3)/mitogen-activated protein kinase kinase 4(MKK4)/c-Jun N-terminal kinase(JNK)pathway in the hippocampus of rats with Parkinson’s disease(PD).Methods Rats were divided into control group,PD model group(PD modeling)and electroacupuncture intervention group(PD modeling+electroacupuncture intervention).The Morris water maze experiment was used to record the escape latency and the staying time on the original platform.Open field experiment was used to record the horizontal and vertical movement scores.HE staining was used to observe the pathological changes of rat hippocampus.TUNEL staining was used to detect cell apoptosis in rat hippocampus.Finally,Western blot assay was used to detect the expression of MAP4K3/MKK4/JNK pathway and apoptosis-related proteins in rat hippocampus.Results Compared with the that in control group,the escape latency of rats in the PD model group was significantly prolonged(P<0.05).Moreover,the staying time on the original platform was also significantly shortened(P<0.05).Consistently,the horizontal movement score and vertical movement score were significantly reduced(P<0.05).Finally,the apoptotic rate and protein expression levels of MAP4K3,p-MKK4/MKK4,p-JNK/JNK,Bax,and cleaved caspase-3 in the hippocampus of rats in the PD model group were significantly increased(P<0.05),accompanied with obvious pathological changes in hippocampus.However,compared with the PD model group,the escape latency of rats in the electroacupuncture intervention group was significantly shorten(P<0.05).The staying time on the original platform was also significantly prolonged(P<0.05);Moreover,the horizontal movement score and vertical movement score were significantly increased(P<0.05).Finally,the apoptotic rate and protein expression levels of MAP4K3,p-MKK4/MKK4,p-JNK/JNK,Bax,and cleaved caspase-3 in the hippocampus of rats were significantly reduced(P<0.05),accompanied with alleviated pathological changes of hippocampus.Conclusion Electroacupuncture can inhibit the activation of MAP4K3/MKK4/JNK pathway,reduce apoptosis and pathological changes in hippocampus of PD rats,and improve cognitive and behavioral functions.