摘要
目的:观察电针干预对创伤性颅脑损伤(TBI)大鼠神经功能、脑组织形态结构、损伤脑组织中半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、(蛋白激酶B)AKT蛋白以及血清神经元特异性烯醇化酶(NSE)表达的影响,探索电针治疗TBI的可能途径。方法:将120只SD大鼠随机分为4组:空白组、假手术组、模型组和电针组。空白组、假手术组各10只;模型组和电针组各50只,进一步分为24 h、3 d、7 d、10 d及14 d 5个亚组,每组各10只。模型组与电针组使用电子颅脑损伤仪,在大鼠颅脑左侧开颅打击,空白组不作处理,假手术组只开颅不打击。选取大鼠“百会”“关元”,右前肢“曲池”“合谷”,右后肢“足三里”“涌泉”,电针组于造模后4 h给予电针干预,至第14天结束。采用改良版神经功能缺损(mNSS)评分评定各组大鼠神经功能缺损情况;苏木精-伊红(HE)染色法观察大鼠脑组织病理形态改变;酶联免疫吸附实验(ELISA)检测大鼠血清中NSE浓度变化;免疫组化法及蛋白质印迹法(Western-blot)检测大鼠损伤脑组织中Caspase-3、AKT蛋白表达情况;荧光定量PCR(qPCR)检测大鼠损伤脑组织中AKT mRNA的变化。结果:与空白组、假手术组比较,造模后24 h、7 d及14 d,模型组大鼠mNSS评分明显升高,差异具有统计学意义(P<0.05);造模后7 d、14 d,与模型组比较,电针组大鼠mNSS评分降低,差异具有统计学意义(P<0.05)。造模后24 h,模型组与电针组损伤脑组织出现病理改变,电针组损伤脑组织出血较模型组轻;造模第3天,模型组与电针组大鼠损伤脑组织病理改变严重;造模后7 d、10 d及14 d,电针组大鼠脑组织形态恢复较同时期模型组好。与空白组和假手术组比较,造模后24 h,模型组、电针组大鼠血清中NSE浓度明显升高,差异具有统计学意义(P<0.01),造模后第3天达到最高,第7天、10天及14天逐渐下降,且电针组浓度始终较同时期模型组低。造模后第24 h、3天、7天、10天及14天,与空白组和假手术组比较,模型组大鼠受损脑组织区域Caspase-3的表达量升高,差异具有统计学意义(P<0.01),AKT表达下降,差异具有统计学意义(P<0.01),与模型组比较,电针组大鼠受损脑组织区域Caspase-3表达量降低,差异具有统计学意义(P<0.01),AKT表达升高,差异具有统计学意义(P<0.01)。造模后24 h、3 d、7 d、10 d及14 d,与空白组和假手术组比较,模型组大鼠损伤脑组织中AKT mRNA的表达量下降,差异具有统计学意义(P<0.01);与模型组比较,电针组大鼠损伤脑组织中AKT mRNA的表达量较高,差异具有统计学意义(P<0.01)。结论:电针干预能明显改善TBI大鼠神经功能、脑组织受损情况;使大鼠血清中NSE浓度降低并上调损伤脑组织中AKT的表达、下调Caspase-3的表达,减轻Caspase-3诱导的内源性途径细胞凋亡,减轻神经元损伤,促进TBI大鼠的恢复。
Objective:To investigate the impact of electro-acupuncture(EA)on the neurological function,brain tissue morphology and structure,the expressions of Caspase-3 and AKT protein in the injured brain tissues,and the concentration of serum NSE in rats suffering from traumatic brain injury(TBI),thus to explore the potential mechanisms of EA in treatment of TBI.Methods:120 SD rats were randomly divided into the blank group,the sham operation group,the model group and the EA group.There were 10 rats in each of the blank group and the sham operation group,and 50 rats in each of the model group and the EA group,which were further divided into five subgroups of 24 h,3 d,7 d,10 d and 14 d,with 10 rats in each subgroup.In the model group and the EA group,a craniocerebral impact was administered to the left side of the rat skull using an electronic brain injury device.The blank group received no operation,while the sham operation group underwent craniotomy without impact.The EA group was intervened with electro-needling Baihui(DU20),Guanyuan(RN4),and Quchi(LI11)and Hegu(LI4)on the right forelimb,as well as Zusanli(ST36)and Yongquan(KI1)on the right hindlimb,4 h after modeling until the end of the 14 d.The neurological deficit was assessed using the mNSS score;pathological changes were observed by means of HE staining;ELISA was used to detect the changes in the serum concentration of NSE;the expressions of Caspase-3 and AKT protein were investigated using immunohistochemistry and Western blot;fluorescence qPCR was applied to the detection of AKT mRNA.Results:The mNSS scores were significantly higher in the model group than those in the blank group and the sham operation group at 24 h,7 d,and 14 d after modeling(P<0.05);the mNSS scores were lower in the EA group than those in the model group at 7 d and 14 d(P<0.05).At 24 h after modeling,the pathological changes appeared in the damaged brain tissues of the model group and the EA group,and the bleeding in the brain tissues was less in the EA group than that in the model group;at the 3 d day after modeling,the pathological changes of the damaged brain tissues were serious in the model group and the EA group;at 7 d,10 d,and 14 d after modeling,the morphology of the brain tissues was better improved in the EA group compared to that in the model group at the same period of time.The serum NSE concentrations were significantly increased in the model group and the EA group compared to those in the blank group and the sham operation group at 24 h after modeling(P<0.01),then concentrations reached the highest level at 3 d after modeling,then declined gradually at 7 d,10 d,and 14 d;the concentrations were lower in the EA group than those in the model group at the same period of time.The expression of Caspase-3 was increased(P<0.01),and the expression of AKT was decreased compared to that in the blank group and the sham operation group at 24 h,3 d,7 d,10 d and 14 d after modeling(P<0.01);the expression of Caspase-3 was decreased(P<0.01),and the expression of AKT was increased in the EA group compared to that in the model group at the same period of time(P<0.01).The expression of AKT mRNA was lower in the model group than that in the blank group and the sham operation group at 24 h,3 d,7 d,10 d,and 14 d after modeling(P<0.01);the expression of AKT mRNA was higher in the EA group than that in the model group at the same period of time(P<0.01).Conclusion:EA intervention can significantly improve the neurological function and brain tissue damage in TBI rats,it decreases serum level of NSE,up-regulates AKT expression and down-regulates Caspase-3 expression in injured brain tissues,attenuates the endogenous pathway apoptosis induced by Caspase-3,alleviates the neuronal damage,and promotes the recovery of TBI rats.
作者
韦晓梅
段荣博
汤阳阳
王瑞辉
吴涛
WEI Xiaomei;DUAN Rongbo;TANG Yangyang;WANG Ruihui;WU Tao(Shaanxi University of Chinese Medicine,Xianyang 712046,China)
出处
《针灸临床杂志》
2025年第7期63-72,共10页
Journal of Clinical Acupuncture and Moxibustion
基金
国家自然科学基金,编号:81904310
陕西省自然科学基金青年项目,编号:2021JQ-729。
