电针联合振动训练对HIE大鼠IκB/NF-κB信号通路与运动机能的影响

Effects of electroacupuncture combined with vibration training on IκB/NF-κB signaling pathway and motor function in HIE rats

目的:研究电针联合振动训练对缺氧缺血性脑病(HIE)大鼠核转录因子抑制蛋白(IκB)/核转录因子κB(NF-κB)信号通路与运动机能的影响。方法:7日龄SD大鼠随机分为假手术组、模型组、电针组、振动组、联合组,每组20只。除假手术组外,其他组均采用改良Rice法建立HIE大鼠模型。造模24 h后,电针组大鼠给予电针干预,针刺穴位为百会、人中和大椎,每次15 min;振动组大鼠进行振动训练,每次15 min;联合组大鼠先后给予电针干预及振动训练。各项干预每日1次,连续2周。末次干预后,采用Bederson评分进行大鼠神经行为学评价,超声诊断仪测量大鼠左后肢腓肠肌横截面积。HE染色后光镜下观察脑组织形态学变化;PCR检测腓肠肌组织IκB、NF-κB p65、Bcl-2、Bax的m RNA表达,Western blot检测腓肠肌组织IκB、NF-κB p65、Bcl-2、Bax的蛋白表达。结果:干预2周后,①电针组、振动组、联合组大鼠的Bederson评分较模型组均显著降低(P<0.05,P<0.01),且联合组评分明显低于振动组和电针组(P<0.05)。②与模型组相比,电针组、振动组及联合组大鼠的左侧腓肠肌横截面积明显增加(P<0.05,P<0.01),且联合组的横截面积较电针组、振动组明显增大(P<0.05,P<0.01)。③HE染色观察显示,模型组大鼠受损脑组织神经细胞排列紊乱,细胞间隙增大、结构疏松,存在空泡样改变,胞体肿胀明显,细胞核萎缩;电针组、振动组和联合组大鼠脑组织病理性损伤均有所减轻,且联合组的改善作用更加显著,脑组织坏死细胞及缺血区炎症浸润明显减少。④与模型组相比,电针组、振动组、联合组腓肠肌组织IκB、NF-κB p65、Bax的m RNA和蛋白表达水平显著下降,Bcl-2的m RNA和蛋白表达明显升高(P<0.05,P<0.01)。与电针组、振动组相比,联合组IκB、NF-κB p65、Bax的m RNA和蛋白表达下调,Bcl-2的m RNA和蛋白表达上调(P<0.05,P<0.01)。结论:电针联合振动训练能够明显改善HIE模型大鼠的神经行为学,减轻脑组织损伤,且疗效优于电针或振动训练单独干预,其作用机制可能与抑制IκB/NF-κB信号通路、抑制骨骼肌细胞凋亡、提高运动机能有关。

Objective:To study the effects of electroacupuncture combined with vibration training on nuclear transcription factor inhibitory protein(IκB)/nuclear transcription factorκB(NF-κB)signaling pathway and motor function in hypoxic-ischemic encephalopathy(HIE)rats.Methods:Seven days old SD rats were randomly divided into the sham operation group,model group,electroacupuncture group,vibration group and combination group,20 rats in each group.Except the sham operation group,HIE model was established by modified Rice method in the other groups.Twenty-four hours after modeling,the electroacupuncture group was treated with electroacupuncture at Baihui,Renzhong and Dazhui acupoints,15 minutes each time;the vibration group was treated with vibration training,15 minutes each time;and the combination group was treated with electroacupuncture and vibration training successively.The treatments were performed once a day for 2 weeks.After the last intervention,the neurobehavior of rats was evaluated by Bederson score,and the cross-sectional area of gastrocnemius muscle of left hindlimb was measured by ultrasonic diagnostic instrument.The morphological changes of brain tissue were observed under light microscope after HE staining.The m RNA expressions of IκB,NF-κB p65,Bcl-2 and Bax in gastrocnemius muscle were detected by PCR.The protein expressions of IκB,NF-κB p65,Bcl-2 and Bax in gastrocnemius muscle were detected by Western blot.Results:After treatment for 2 weeks,①The Bederson score of rats in the electroacupuncture group,vibration group and combination group was significantly lower than that in the model group(P<0.05,P<0.01),and the score of the combination group was significantly lower than that of the vibration group and the electroacupuncture group(P<0.05).②Compared with the model group,the cross-sectional area of left gastrocnemius muscle in the electroacupuncture group,vibration group and combination group was increased significantly(P<0.05,P<0.01).The cross-sectional area of the combination group was significantly larger than that of the electroacupuncture group and the vibration group(P<0.05,P<0.01).③HE staining showed that the arrangement of nerve cells in the damaged brain tissue of the model group was disordered,the intercellular space was enlarged,the structure was loose,and the vacuole-like changes,obvious cell body swelling and nuclear atrophy were observed.The pathological injury of brain tissue in the electroacupuncture group,vibration group and combination group was alleviated,and the improvement in the combination group was more significant,and the necrotic cells of brain tissue and inflammatory infiltration in ischemic area were significantly reduced.④Compared with the model group,the m RNA and protein expression levels of IκB,NF-κB p65 and Bax in gastrocnemius muscle of the electroacupuncture group,vibration group and combination group were significantly decreased,and the m RNA and protein expression levels of Bcl-2 were significantly increased(P<0.05,P<0.01).Compared with the electroacupuncture group and vibration group,the m RNA and protein expressions of IκB,NF-κB p65 and Bax were down-regulated in the combination group,and the m RNA and protein expressions of Bcl-2 were up-regulated(P<0.05,P<0.01).Conclusion:Electroacupuncture combined with vibration training can significantly improve neurobehavior and reduce brain tissue injury in HIE model rats,and the effect is better than electroacupuncture or vibration training alone.Its mechanism may be related to suppressing IκB/NF-κB signaling pathway,inhibiting cell apoptosis of skeletal muscle and improving motor function.