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
李卓航 1王栋 1王艳秋 1戚明珠 1黄荷兰 1林娜 1苏晓慧 1孔祥英1
作者信息
- 1. 中国中医科学院 中药研究所,北京 100700
- 折叠
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 oxidoreduc-tase 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 50 μg·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.
Abstract
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 mode-ling,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-regula-ted the expression of Nrf2,CXCR4,and SDF-1.In addition,TMP promoted the migration of C17.2 cells and up-regulated the expres-sion of p62,Nrf2,HO-1,and NQO1 in a time-and dose-dependent manner.The expression was the highest at the time point of 12 h in the TMP(50 μg·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Key words
middle cerebral artery occlusion/tetramethylpyrazine/migration of neural stem cells/Nrf2/HO-1/CXCR4 pathway引用本文复制引用
基金项目
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)
国家自然科学基金面上项目(81673630)
中国中医科学院科技创新工程重大攻关项目(CI2021A04610)
出版年
2024
NETL
NSTL
万方数据