川芎嗪激活Nrf2/HO-1/CXCR4通路调控神经干细胞迁移干预缺血再灌注大鼠的作用机制  

Mechanism of tetramethylpyrazine intervention with ischemia-reperfusion rats based on Nrf2/HO-1/CXCR4 pathway through regulating neural stem cell migration

作　　者：李卓航 王栋 王艳秋 戚明珠 黄荷兰 林娜[1] 苏晓慧[1] 孔祥英[1] LI Zhuo-hang;WANG Dong;WANG Yan-qiu;QI Ming-zhu;HUANG He-lan;LIN Na;SU Xiao-hui;KONG Xiang-ying(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国中药杂志》2024年第9期2308-2315,共8页China Journal of Chinese Materia Medica

基　　金：北京市自然科学基金面上项目(7222291);国家自然科学基金面上项目(82074048,81673630);中国中医科学院科技创新工程重大攻关项目(CI2021A04610)。

摘　　要：以核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/血红素加氧酶1(heme oxygenase 1,HO-1)/趋化因子C-X-C-基元受体4(C-X-C motif chemokine receptor 4,CXCR4)通路为切入点,探讨川芎嗪(tetramethylpyrazine,TMP)干预大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)模型大鼠脑内神经干细胞(neural stem cells,NSCs)迁移的作用机制。SD大鼠分为假手术组、模型组、TMP 20 mg·kg^(-1)组和TMP 40 mg·kg^(-1)组;通过神经功能缺失评分评价神经功能损伤;免疫荧光法检测脑组织5-溴脱氧尿苷(5-bromodeoxyuridine,BrdU)/双皮质素(doublecortin,DCX)双标细胞;观察TMP对C17.2细胞迁移的影响;蛋白免疫印迹法检测脑组织和C17.2细胞中Nrf2、HO-1、p62、醌氧化还原酶1[NAD(P)H quinone oxidoreductase 1,NQO1]、基质细胞衍生因子1(stromal cell-derived factor 1,SDF-1)、CXCR4蛋白表达。结果显示,大鼠脑缺血7、21 d后,神经功能损伤评分及BrdU+/DCX+细胞显著升高,脑组织中Nrf2及CXCR4表达均显著升高;与模型组相比,TMP 40 mg·kg-1可显著降低神经功能评分,进一步升高BrdU+/DCX+细胞数量及Nrf2、CXCR4及SDF-1表达;此外TMP可以显著促进C17.2细胞迁移,且时间以及剂量依赖性提高p62、Nrf2、HO-1、NQO1表达,TMP 50μg·mL^(-1)给药12 h表达量最高(P<0.01)。综上,TMP可通过活化Nrf2/HO-1/CXCR4通路,促进NSCs迁移从而发挥抗缺血再灌注损伤作用。该研究为TMP在缺血性脑卒中疾病中的应用提供了实验支持。This study aims to decipher the mechanism of tetramethylpyrazine(TMP)in regulating the migration of neural stem cells(NSCs)in the rat model of middle cerebral artery occlusion(MCAO)via the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/C-X-C motif chemokine receptor 4(CXCR4)pathway.SD rats were randomized into sham,MCAO(model),and tetramethylpyrazine(TMP,20 mg·kg^(-1)and 40 mg·kg^(-1))groups.The neurological impairment was assessed by the modified neu-rological severity score(mNSS).The immunofluorescence assay was employed to detect the cells stained with both 5-bromodeoxyuri-dine(BrdU)and doublecortin(DCX)in the brain tissue.The effect of TMP on the migration of C17.2 cells was observed.Western blot was employed to determine the protein levels of Nrf2,HO-1,p62,NAD(P)H quinone oxidoreductase 1(NQO1),stromal cell derived factor 1(SDF-1),and CXCR4 in the brain tissue and C17.2 cells.The results showed that after 7 days and 21 days of modeling,the mNSS and BrdU*/DCX*cells were increased,and the expression of Nrf2 and CXCR4 in the brain tissue was up-regulated.Compared with the model group,TMP(40 mg·kg^(-1))reduced the mNSS,increased the number of BrdU*/DCX*cells,and up-regulated the expression of Nrf2,CXCR4,and SDF-1.In addition,TMP promoted the migration of C17.2 cells and up-regulated the expression of p62,Nrf2,HO-1,and NQ01 in a time-and dose-dependent manner.The expression was the highest at the time point of 12 h in the TMP(50μg·mL^(-1))group(P<0.01).In conclusion,TMP activates the Nrf2/HO-1/CXCR4 pathway to promote the migration of NSCs to the ischemic area,thus exerting the therapeutic effect on the ischemia-reperfusion injury.This study provides experimental support for the application of TMP in ischemic stroke.

关 键 词：大脑中动脉阻塞 川芎嗪 神经干细胞迁移 Nrf2/HO-1/CXCR4通路 

分 类 号：R285.5[医药卫生—中药学]