电针刺激头部运动区对脑瘫大鼠脑损伤及JAK2/STAT3信号通路的影响

Electroacupuncture stimulation of the head relieves motor function impairment in rats modeling cerebral palsy

目的观察电针刺激头部运动区对脑瘫大鼠脑损伤及JAK2/STAT3信号通路的影响。方法将30只SD大鼠随机分为假手术组、模型组及电针刺激组,每组10只大鼠。将模型组及电针刺激组大鼠制成脑瘫动物模型,假手术组大鼠术中不结扎左颈总动脉。于造模后24 h及21 d时采用BBB运动功能评分评价各组大鼠运动功能情况,并于造模后21 d时处死各组大鼠并提取脑组织。采用苏木素-伊红染色观察各组大鼠脑皮质病理学改变;采用荧光定量PCR法检测凋亡基因B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关x蛋白(Bax)、半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3)mRNA表达水平;采用ELISA法检测肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、干扰素-γ(IFN-γ)及氧化应激指标超氧化物歧化酶(SOD)、总抗氧化力(T-AOC)、丙二醛(MDA)含量;采用Western blot法检测JAK2、STAT3、p-JAK2及p-STAT3表达水平。结果造模后21 d时模型组大鼠BBB运动功能评分、脑组织中Bcl-2 mRNA表达、SOD、T-AOC含量均显著低于假手术组水平(P<0.05),Caspase-3 mRNA表达、TNF-α、IL-1β、IFN-γ、MDA含量、p-JAK2和p-STAT3表达水平均明显高于假手术组水平(P<0.05);电针刺激组大鼠BBB运动功能评分、Bcl-2 mRNA表达、SOD、T-AOC含量均显著高于模型组水平(P<0.05),Caspase-3 mRNA表达、TNF-α、IL-1β、IFN-γ、MDA含量、p-JAK2及p-STAT3表达均明显低于模型组水平(P<0.05)。结论电针刺激头部运动区可缓解脑瘫模型大鼠脑损伤,改善肢体运动功能,其作用机制可能与抑制JAK2/STAT3信号通路激活,进而减轻细胞凋亡、炎症反应及氧化应激反应有关。

Objective To observe any effect of electroacupuncture in the head motor area on motor impairment and JAK2/STAT3 signaling in rats modeling cerebral palsy.Methods Thirty Sprague-Dawley rats were randomly divided into a sham operation group,a model group and an electroacupuncture group,each of 10.A model of cerebral palsy was induced in the model and electroacupuncture groups,while the sham operation group was not ligated in a sham operation.The motor functioning of all of the rats was evaluated 24h and 21d after the modeling using the Basso,Beattie Bresnahan locomotor rating scale(BBB scale).At 21d all of the rats were sacrificed and their brain tissues were collected.Hematoxylin-eosin staining was used to observe any pathological changes in the cerebral cortex,and fluorescence quantitative PCRs were employed to detect the mRNA expression of apoptotic gene B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2-associated x-protein(Bax),and cysteine-aspartate protein hydrolase-3(Caspase-3).ELISA was used to detect the mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interferon-γ(IFN-γ),and the oxidative stress indicators superoxide dismutase(SOD),total antioxidant capacity(T-AOC),and malondialdehyde(MDA).Western blotting was applied to detect the expression of JAK2,STAT3,p-JAK2 and p-STAT3.Results The average BBB motor function score,Bcl-2 mRNA expression,SOD and T-AOC of the model group were all significantly lower than that of the sham-operated group at 21d.The average expression of Caspase-3 mRNA,TNF-α,IL-1βand IFN-γ,were significantly greater as were MDA content,p-JAK2,and p-STAT3 levels.The average BBB motor function score of the electroacupuncture group was significantly higher than the model group′s average,but caspase-3 mRNA expression,TNF-α,IL-1β,IFN-γ,MDA,p-JAK2 and p-STAT3 were all significantly lower.Conclusion Electroacupuncture applied to the head motor area relieves motor function impairment in rats modeling cerebral palsy.That may be related to inhibition of the JAK2/STAT3 pathway′s activation resulting in less apoptosis,inflammation and oxidative stress.