摘要
目的研究电针(Electroacupuncture,EA)是否通过下调谷氨酸羧肽酶Ⅱ(Glutamate carboxypeptidaseⅡ,GCPⅡ)基因表达减轻大鼠创伤性脑损伤(Traumatic brain injury,TBI)。方法将大鼠分为Sham组(n=10)、TBI组(n=11)、TBI+EA组(n=11)、TBI+EA+GCPⅡ-NC组(n=11)和TBI+EA+GCPⅡ-OE组(n=11)。Sham组大鼠进行假手术,其他组大鼠使用电子颅脑损伤仪进行TBI造模。Sham组和TBI组大鼠正常饲养,其他组大鼠均进行电针治疗,共治疗14 d。TBI+EA+GCPⅡ-NC组和TBI+EA+GCPⅡ-OE组大鼠在电针治疗基础上,分别脑室注射GCPⅡ-NC和GCPⅡ-OE。治疗结束后,通过神经功能评分和Morris水迷宫实验评价大鼠认知功能。对大鼠脑组织进行HE和TUNEL染色。检测各组大鼠脑组织的含水量、谷氨酸(Glutamate,Glu)含量、钙离子(Ca^(2+))浓度、超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(Catalase,CAT)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)和丙二醛(Malondialdehyde,MDA)水平。通过qRT-PCR或Western blot检测脑组织中GCPⅡ、Bax、Bcl2和cleaved caspase-3的mRNA或蛋白表达水平。结果与Sham组比较,TBI组大鼠脑组织中GCPⅡ的mRNA和蛋白表达水平升高,神经功能评分和逃避潜伏期升高,穿越平台次数降低,含水量升高,皮层出现明显损伤,Glu含量升高,Ca^(2+)浓度升高,SOD、CAT和GSH-Px水平降低,MDA水平升高,TUNEL阳性率升高,Bax和cleaved caspase-3蛋白表达水平升高,Bcl2蛋白表达水平显著降低(P<0.05)。与TBI组比较,TBI+EA组大鼠脑组织中GCPⅡ的mRNA和蛋白表达水平降低,神经功能评分和逃避潜伏期降低,穿越平台次数升高,含水量降低,皮层形态明显改善,Glu含量降低,Ca^(2+)浓度降低,SOD、CAT和GSH-Px水平升高,MDA水平降低,TUNEL阳性率降低,Bax和cleaved caspase-3蛋白表达水平降低,Bcl2蛋白表达水平显著升高(P<0.05)。与TBI+EA组和TBI+EA+GCPⅡ-NC组比较,TBI+EA+GCPⅡ-OE组大鼠脑组织中GCPⅡ的mRNA和蛋白表达水平升高,神经功能评分和逃避潜伏期升高,穿越平台次数降低,含水量升高,皮层损伤加重,Glu含量升高,Ca^(2+)浓度升高,SOD、CAT和GSH-Px水平降低,MDA水平升高,TUNEL阳性率升高,Bax和cleaved caspase-3蛋白表达水平升高,Bcl2蛋白表达水平显著降低(P<0.05)。结论电针通过下调GCPⅡ基因表达降低了TBI大鼠脑组织中的谷氨酸含量,从而抑制了脑组织钙超载、氧化应激和神经元凋亡,进而改善了认知功能并减轻了脑损伤。
Objective To study whether electroacupuncture(EA)attenuates traumatic brain injury(TBI)by downregulating glutamate carboxypeptidaseⅡ(GCPII)gene expression in rats.Methods Rats were divided into Sham group(n=10),TBI group(n=11),TBI+EA group(n=11),TBI+EA+GCPII-NC group(n=11)and TBI+EA+GCPII-OE group(n=11).Sham group rats underwent sham surgery,while other groups rats underwent TBI modeling using an electronic brain injury instrument.The rats in Sham group and TBI group were fed normally,and the rats in other groups were treated with electroacupuncture for 14 days.On the basis of electroacupuncture treatment,rats in TBI+EA+GCPII-NC group and TBI+EA+GCPII-OE group were injected with GCPII-NC and GCPII-OE,respectively.After treatment,the cognitive function of rats was evaluated by neurological function score and Morris water labyrinth task.HE and TUNEL staining were performed on brain tissue.The water content,glutamate(Glu)content,calcium(Ca^(2+))concentration,superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in brain tissue of rats in each group were measured.The mRNA or protein expression of GCPII,Bax,Bcl2 and cleaved caspase-3 in brain tissue were detected by qRT-PCR or Western blot.Results Compared with Sham group,the expression of mRNA and protein of GCPII in TBI group increased,the neurological function score and escape latency increased,the number of crossing the platform decreased,the water content increased,the cortex showed obvious damage,the content of Glu increased,the concentration of calcium increased,the levels of SOD,CAT and GSH-Px decreased,the level of MDA increased,the positive rate of TUNEL increased,the expression of Bax and cleaved caspase-3 protein increased,and the expression of Bcl2 protein decreased significantly(P<0.05).Compared with TBI group,the expression level of mRNA and protein of GCPII in TBI+EA group decreased,the neurological function score and escape latency decreased,the number of crossing platform increased,the water content decreased,the morphology of cortex improved obviously,the content of Glu decreased,the concentration of calcium decreased,the level of SOD,CAT and GSH-Px increased,the level of MDA decreased,the positive rate of TUNEL decreased,the expression level of Bax and cleaved caspase-3 protein decreased,and the expression level of Bcl2 protein increased significantly(P<0.05).Compared with TBI+EA group and TBI+EA+GCPII-NC group,the expression level of mRNA and protein of GCPII in TBI+EA+GCPII-OE group increased,neurological function score and escape latency increased,the number of crossing platform decreased,water content increased,cortical injury aggravated,Glu content increased,calcium concentration increased,SOD,CAT and GSH-Px levels decreased,MDA level increased,TUNEL positive rate increased,Bax and cleaved caspase-3 protein expression increased,and the expression of Bcl2 protein decreased significantly(P<0.05).Conclusion By downregulating the expression of GCPII gene,electroacupuncture reduced the content of glutamate in the brain tissue of TBI rats,thereby inhibiting calcium overload,oxidative stress and neuronal apoptosis in the brain tissue,and thereby improving cognitive function and alleviating brain injury.
作者
韦晓梅
段荣博
汤阳阳
吴涛
WEI Xiaomei;DUAN Rongbo;TANG Yangyang;WU Tao(School of Acupuncture-Moxibustion and Tuina,Shaanxi University of Chinese Medicine,Xianyang 712046,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第7期1887-1897,共11页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会青年科学基金项目(81904310):电针基于AMPK/mTOR通路调节颅脑损伤后自噬与康复新机制,负责人:吴涛
陕西省科学技术厅陕西省自然科学基金委青年项目(2021JQ-729):基于细胞自噬研究电针调控颅脑损伤康复新机制,负责人:吴涛。
关键词
创伤性脑损伤
电针
谷氨酸羧肽酶Ⅱ
谷氨酸
钙超载
氧化应激
神经元凋亡
Traumatic brain injury
Electroacupuncture
Glutamate carboxypeptidaseⅡ
Glutamate
Calcium overload
Oxidative stress
Neuronal apoptosis
