基于OPN/IGF-1/mTOR探讨左归丸对缺血性脑卒中小鼠神经功能恢复作用及其机制  

Effect and mechanism of Zuogui Pills on neural function recovery in ischemic stroke mice based on OPN/IGF-1/mTOR

作　　者：刘燕 高春辰 厉励 吴旦 丛羽君 冯清华 吴明华[1] 李文磊 LIU Yan;GAO Chun-chen;LI Li;WU Dan;CONG Yu-jun;FENG Qing-hua;WU Ming-hua;LI Wen-lei(the Affiliated Hospital of Nanjing University of Chinese Medicine/Jiangsu Province Hospital of Chinese Medicine,the First School of Clinical Medicine of Nanjing University of Chinese Medicine,Nanjing 210029,China;Department of Medical Genetics and Developmental Biology,Air Force Medical University,Xi′an 710032,China)

机构地区：[1]南京中医药大学附属医院/江苏省中医院南京中医药大学第一临床医学院,江苏南京210029 [2]空军军医大学医学遗传与发育生物学教研室,陕西西安710032

出　　处：《中国中药杂志》2023年第19期5250-5258,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81973794);江苏省中医药管理局项目(JD201803,ZT202102);江苏省研究生科研与实践创新计划项目(KYCX221902)。

摘　　要：探讨左归丸促进缺血性脑卒中小鼠神经组织修复和功能恢复作用及其机制。雄性C57BL/6J小鼠随机分为5组:假手术组、模型组、左归丸低剂量组(3.5 g·kg^(-1))、左归丸中剂量组(7 g·kg^(-1))、左归丸高剂量组(14 g·kg^(-1)),每组15只。采用光化学栓塞法建立缺血性脑卒中模型。分别于造模前和给药第7、14、21、28天进行行为学检测。在造模3 d后进行2,3,5-氯化三苯基四氮唑(TTC)染色,给药28 d后进行影像学T2加权成像(T2-weighted imaging,T2WI)检测、扩散加权成像(diffusion-weighted imaging,DWI)检测,评估模型损伤情况。苏木素-伊红(hematoxylin-eosin,HE)染色观察脑皮质区组织学情况。通过免疫荧光和免疫蛋白印迹观察轴突标记蛋白(myelin basic protein,MBP)、轴突生长相关蛋白43(growth-associated protein 43,GAP43)、细胞内在活力相关通路哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)及其下游磷酸化S6核糖体蛋白(phosphorylated s6 ribosomal protein,p-S6)以及机制相关蛋白骨桥蛋白(osteopontin,OPN)、胰岛素样生长因子1(insulin-like growth factor 1,IGF-1)的相对表达量。左归丸治疗组对缺血性脑卒中小鼠神经功能的损伤有一定的恢复作用,TTC染色显示感觉运动皮质区出现白色梗死灶,T2WI影像学上观察到感觉运动皮质区坏死,形成囊腔;脑组织损伤较轻、瘢痕较少、毛细血管增多,神经元轴突较模型组增多、神经元活力较强。与模型组相比,左归丸治疗组GAP43、OPN、IGF-1、mTOR蛋白表达明显增多。综上,左归丸能促进缺血性脑梗死小鼠神经功能恢复,促进轴突生长,其机制可能与激活OPN/IGF-1/mTOR信号轴有关。To explore the effect and mechanism of Zuogui Pills in promoting neural tissue recovery and functional recovery in mice with ischemic stroke.Male C57BL/6J mice were randomly divided into a sham group,a model group,and low-,medium,and high-dose Zuogui Pills groups(3.5,7,and 14 g·kg^(-1)),with 15 mice in each group.The ischemic stroke model was established using photochemical embolization.Stiker remove and irregular ladder walking behavioral tests were conducted before modeling and on days 7,14,21,and 28 after medication.Triphenyl tetrazolium chloride(TTC) staining was performed on day 3 after modeling,and T2-weighted imaging(T2WI) and diffusion-weighted imaging(DWI) were performed on day 28 after medication to evaluate the extent of brain injury.Hematoxylin-eosin(HE) staining was performed to observe the histology of the cerebral cortex.Axonal marker proteins myelin basic protein(MBP),growth-associated protein 43(GAP43),mammalian target of rapamycin(mTOR),and its downstream phosphorylated s6 ribosomal protein(p-S6),as well as mechanism-related proteins osteopontin(OPN) and insulin-like growth factor 1(IGF-1),were detected using immunofluorescence and Western blot.Zuogui Pills had a certain restorative effect on the neural function impairment caused by ischemic stroke in mice.TTC staining showed white infarct foci in the sensory-motor cortex area,and T2WI imaging revealed cystic necrosis in the sensory-motor cortex area.The Zuogui Pills groups showed less brain tissue damage,fewer scars,and more capillaries.The number of neuronal axons in those groups was higher than that in the model group,and neuronal activity was stronger.The expression of GAP43,OPN,IGF-1,and mTOR proteins in the Zuogui Pills groups was higher than that in the model group.In summary,Zuogui Pills can promote the recovery of neural function and axonal growth in mice with ischemic stroke,and its mechanism may be related to the activation of the OPN/IGF-1/mTOR signaling pathway.

关 键 词：左归丸 缺血性脑卒中 神经功能恢复 OPN IGF-1 MTOR 

分 类 号：R285.5[医药卫生—中药学]