补阳还五汤联合内皮祖细胞移植减少脑缺血再灌注损伤的作用研究  

Study on the effect of Buyang Huanwu Tang combined with endothelial progenitor cell transplantation on reducing cerebral ischemia-reperfusion injury

作　　者：侯伯南 吴忧 徐彬 梁顺利 张荣博 徐林胜 HOU Bonan;WU You;XU Bin;LIANG Shunli;ZHANG Rongbo;XU Linsheng(Department of Neurology,Second Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine,Hangzhou 310000,China)

机构地区：[1]浙江中医药大学附属第二医院神经内科,杭州310000

出　　处：《环球中医药》2024年第12期2481-2487,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(82104762)。

摘　　要：目的观察补阳还五汤联合内皮祖细胞(endothelial progenitor cells,EPCs)移植对脑缺血再灌注损伤的作用。方法通过5周龄的C57BL/6小鼠取材培育EPCs。将30只雄性C57BL/6小鼠(6~8周龄)随机分为两部分,一部分(6只)作为正常对照组,另一部分(24只)构建大脑中动脉闭塞(middle cerebral artery occlusion/reperfusion,MCAO/R)模型。将造模成功的小鼠随机分为4组,使用EPCs移植和(或)补阳还五汤干预,根据干预的方法命名为MCOA/R组、EPCs组、补阳还五汤组和补阳还五汤联合EPCs组,每组6只。造模24小时后通过尾静脉注射EPCs和(或)补阳还五汤灌胃干预,疗程共7天。运用神经功能评分、氯化三苯基四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)染色评估脑梗死面积,免疫组化分析Ki67的表达,比色法检测血清超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)水平,免疫蛋白印迹(Western blot,WB)法检测小鼠脑组织中醌氧化还原酶1[NAD(P)H quinone oxidoreductase 1,NQO1]、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2 associated X,Bax)的表达。结果与MCOA/R组相比,各治疗组能明显改善小鼠的神经功能评分(P<0.05),减少脑梗死面积(P<0.01),增加Ki67表达(P<0.01),增加SOD水平和减少MDA水平(P<0.01),增加NQO1的表达(P<0.01),以及增加Bcl-2、减少Bax的表达水平(P<0.05)。其中补阳还五汤联合EPCs组干预作用更显著。结论补阳还五汤通过抑制氧化应激,减少凋亡,促进EPCs增殖和分化,减少脑梗死面积,与EPCs联合发作协同增效作用减少脑缺血再灌注损伤。Objective To investigate the effect of Buyang Huanwu Tang combined with endothelial progenitor cells(EPCs)on cerebral ischemia-reperfusion.Methods EPCs were first cultured from C57BL/6 mice at 5 weeks of age.Then,30 C57BL/6 mice(6~8 weeks of age)were divided into two groups:a normal control group(6 mice)and a MCOA/R model was constructed and interventions were applied with EPCs transplantation and/or Buyang Huanwu Tang.The successfully modeled mice were randomly divided into MCOA/R group,EPCs group,Buyang Huanwu Tang group,and Buyang Huanwu Tang combined with EPCs group,with 6 mice in each group.24 hours after model establishment,EPCs and/or Buyang Huanwu Tang were injected into the mice via tail vein and administered orally for a total duration of 7 sessions.The infarct area was evaluated by neurological function scoring,staining with 2,3,5-triphenyltetrazolium chloride(TTC),immunohistochemical analysis of Ki67 expression,colorimetric detection of serum superoxide dismutase(SOD)and malondialdehyde(MDA)levels,and Western blotting(WB)detection of NAD(P)H quinone oxidoreductase 1(NQO1),Bcl-2,and Bax expression in mouse brain tissue.Results Compared with the MCOA/R group,each treatment group significantly improved the neurological function score of mice(P<0.05),reduced cerebral infarction area(P<0.01),increased Ki67 expression(P<0.01),increased SOD levels and decreased MDA levels(P<0.01),increased NQO1 expression(P<0.01),and increased Bcl-2 and decreased Bax expression levels(P<0.05).The intervention effect of Buyang Huanwu Tang combined with EPCs was more significant.Conclusion Buyang Huanwu Tang can inhibit oxidative stress,reduce apoptosis,promote the proliferation and differentiation of EPCs,reduce the area of cerebral infarction,and has a synergistic effect in reducing cerebral ischemia-reperfusion injury when combined with EPCs.

关 键 词：补阳还五汤 内皮祖细胞 缺血再灌注 氧化应激 凋亡 血脑屏障 

分 类 号：R285.5[医药卫生—中药学]