脑震宁颗粒调节三重脑震荡大鼠线粒体能量代谢的机制研究  

Mechanism of Naozhenning granules in regulating mitochondrial energy metabolism in multiple cerebral concussion rats

作　　者：高丽 赵乐 魏楠楠 武立雅 王恬恬 张唯依 王永辉 GAO Li;ZHAO Le;WEI Nannan;WU Liya;WANG Tiantian;ZHANG Weiyi;WANG Yonghui(Basic MedicalCollege,Shanxi University of Chinese Medicine,Shanxi Jinzhong 030619,China)

机构地区：[1]山西中医药大学基础医学院,山西晋中030619

出　　处：《中国药房》2024年第9期1050-1056,共7页China Pharmacy

基　　金：国家自然科学基金项目(No.82104972);山西省卫生健康委员会中医药重点研究室建设项目(No.zyyyjs2024023);山西省中医药管理局科研课题(No.2022ZYYC267);山西中医药大学博士科研启动基金项目(No.2022BK19);山西中医药大学科技创新能力培育计划(No.2023PY-YS-03)。

摘　　要：目的探究脑震宁颗粒调节三重脑震荡(MCC)模型大鼠海马组织线粒体能量代谢的作用机制。方法取SPF级Wistar大鼠,通过“自由落体撞击法”制备MCC模型。将造模成功的大鼠分为模型组、吡拉西坦组和脑震宁颗粒低、中、高剂量组,另设正常组,每组8只。各给药组大鼠按吡拉西坦组0.324 g/kg,脑震宁颗粒低、中、高剂量组2.25、4.5、9 g/kg的剂量灌服相应药物,正常组和模型组给予等体积生理盐水,每天1次,连续14 d。检测大鼠运动探索能力和学习记忆能力;检测大鼠海马组织三磷酸腺苷(ATP)含量;观察大鼠海马组织线粒体结构变化;检测大鼠海马组织线粒体动力蛋白相关蛋白1(Drp1)、线粒体裂变蛋白1(Fis1)、线粒体融合蛋白1(Mfn1)、视神经萎缩蛋白1(Opa1)的荧光强度;检测大鼠海马组织过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)、核呼吸因子1(NRF-1)、线粒体转录因子A(TFAM)、Wnt-3a、β-连环蛋白(β-catenin)的蛋白表达水平。结果与正常组比较,模型组大鼠的运动总路程、中央格进入次数、直立次数,新物体识别指数,海马组织线粒体ATP含量,Mfn1、Opa1的荧光强度及PGC-1α、NRF-1、TFAM、Wnt-3a、β-catenin蛋白表达水平均显著降低(P<0.01),静止时间和海马组织Drp1、Fis1的荧光强度均显著升高(P<0.01);透射电镜结果显示,海马组织线粒体明显肿胀,嵴大量断裂、减少,部分线粒体膜内髓样变。与模型组比较,各给药组大鼠以上指标水平/含量均有不同程度的逆转,大部分差异有统计学意义(P<0.05或P<0.01);透射电镜结果显示,海马组织线粒体肿胀程度减轻,嵴少量断裂、减少,粗面内质网局部区域模糊断裂。结论脑震宁颗粒可以提高MCC模型大鼠的运动探索和学习记忆能力,修复神经元损伤,发挥神经保护作用;其机制可能与激活Wnt/β-catenin信号通路,维持线粒体融合分裂平衡,促进线粒体生物合成相关�OBJECTIVE To explore the mechanism of Naozhenning granules in regulating mitochondrial energy metabolism in hippocampal tissue of multiple cerebral concussion(MCC)model rats.METHODS SPF grade Wistar rats were used to prepare MCC models using the“free fall impact method”.The successfully modeled rats were divided into model group,piracetam group,and Naozhenning granule low-dose,medium-dose and high-dose groups,and a normal group was also set up,with 8 rats in each group.Rats in each treatment group orally administered corresponding drugs at doses of 0.324 g/kg for the piracetam group and 2.25,4.5 and 9 g/kg for the Naozhenning granule low-dose,medium-dose and high-dose groups;the normal group and model group were given equal volumes of normal saline;once a day,for 14 consecutive days.The motor exploration ability,learning and memory ability of rats were tested;the adenosine triphosphate(ATP)content in the hippocampal tissue of rat was detected;the changes in the mitochondrial structure of hippocampal tissue was observed;the fluorescence intensity of mitochondrial dynaminrelated protein 1(Drp1),mitochondrial fission protein 1(Fis1),mitochondrial fusion 1(Mfn1),and optic atrophy protein 1(Opa1)were detected in the hippocampal tissue of rat;the protein expression levels of peroxisome proliferator activated receptor gamma coactivator-1α(PGC-1α),nuclear respiratory factor-1(NRF-1),mitochondrial transcription factor A(TFAM),Wnt-3a,β-catenin in hippocampal tissue of rat were detected.RESULTS Compared with the normal group,the total exercise distance,number of central grid entries,number of upright positions,new object recognition index,mitochondrial ATP content,fluorescence intensity of Mfn1 and Opa1,the protein expression levels of PGC-1α、NRF-1、TFAM、Wnt-3a、β-catenin in the model group were significantly reduced(P<0.01),while the rest time and fluorescence intensity of Drp1 and Fis1 in hippocampal tissue were significantly increased(P<0.01).The results of transmission electron microscopy showed that the

关 键 词：脑震宁颗粒 三重脑震荡 线粒体 WNT/Β-CATENIN信号通路 海马组织 神经保护 

分 类 号：R965[医药卫生—药理学] R285.5[医药卫生—药学]