芪芎左归复方干预骨髓间充质干细胞外泌体对缺血性脑卒中大鼠血脑屏障通透性的影响及抗衰老作用

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中文摘要：探讨芪芎左归复方干预骨髓间充质干细胞(bone marrow stem cells，BMSCs)外泌体对衰老大脑中动脉栓塞(middle cerebral artery occlusion，MCAO)大鼠血脑屏障(blood-brain barrier，BBB)的修复作用及抗衰老的作用机制。采用D-半乳糖致衰老法联合线栓法建立衰老MCAO复合模型，实验设青年假手术组、衰老假手术组、模型组、外泌体组、中药干预的外泌体组，使用β-半乳糖苷酶(SA-β-gal)染色联合TTC染色评估模型的建立情况，术后2 h及脑缺血再灌注后48、96 h外泌体组及中药干预的外泌体组分别于尾静脉注射相应的外泌体。造模成功后2 h、3 d、7 d进行神经功能缺损评分;第7天通过新物体识别实验检测认知功能、磁共振观察脑梗死体积;伊文思蓝染色评价BBB通透性;蛋白免疫印迹法(Western blot)检测脑组织中紧密连接蛋白ZO-1、Occludin的表达;酶联免疫吸附法(ELISA)检测血清基质金属蛋白酶(MMP)-9、MMP-2水平;SA-β-gal染色检测衰老细胞情况;反转录实时荧光定量PCR(RT-qPCR)检测p21 mRNA的表达，实时荧光定量PCR(qPCR)检测端粒相对长度。结果显示，芪芎左归复方干预BMSCs外泌体能显著降低衰老MCAO大鼠脑梗死体积，改善神经功能缺损评分，提高认知功能;并能抑制MMP-2、MMP-9表达，促进紧密连接蛋白ZO-1、Occludin的表达，降低BBB的通透性;同时还能降低p21 mRNA的表达，延长端粒长度，延缓细胞衰老。综上，芪芎左归复方干预BMSCs外泌体可能通过降低MMP-2、MMP-9含量，提高ZO-1、Occludin表达水平来修复BBB，同时其抗衰老机制可能与降低p21 mRNA的表达、限制端粒缩短有关。

外文标题：Effects of Qixiong Zuogui Compound Prescription on blood-brain barrier permeability of rats with ischemic stroke by interfering in exosomes of bone marrow stem cells and its anti-aging effects

外文摘要：This study explores the reparative effect of Qixiong Zuogui Compound Prescription(QXZG)intervention on the blood-brain barrier(BBB)in the aging brain with middle cerebral artery occlusion(MCAO)in rats mediated by bone marrow stem cells(BMSCs)-derived exosomes,as well as its anti-aging mechanism.An aging MCAO composite model was established using D-galactose-induced aging combined with line embolism.Rats were divided into young sham surgery group,aging sham surgery group,model group,exosome group,and exosome with traditional Chinese medicine(TCM)intervention group.SA-β-gal staining combined with TTC staining were used to evaluate the model establishment.Exosomes group or exosomes with TCM intervention group were injected via the tail vein 2 h post-surgery and 48 and 96 h post-ischemia reperfusion.Neurological deficit scores were assessed at 2 h,3d,and 7 d post-modeling.Cognitive function was evaluated using novel object recognition tests on the 7th day,and brain infarct volume was observed via MRI.Evans Blue staining was used to evaluate BBB permeability.Western blot was performed to detect the expression of tight junction proteins ZO-1 and Occludin in brain tissue.ELISA was used to measure serum matrix metalloproteinase(MMP)-9 and MMP-2 levels.SA-β-gal staining was used to assess cellular senescence.RT-qPCR was performed to detect p21 mRNA expression,and qPCR was used to measure relative telomere length.The results showed that QXZG intervention-derived BMSCs exosomes signifi-cantly reduced brain infarct volume in aging MCAO rats,improved neurological deficit scores,and enhanced cognitive function.These exosomes suppressed MMP-2 and MMP-9 expression,promoted he expression of tight junction proteins ZO-1 and Occludin,reduced BBB permeability,lowered p21 mRNA expression,elongated telomeres,and delayed cell senescence.In conclusion,this study con-firms that QXZG intervention-derived BMSC exosomes may repair the BBB by reducing MMP-2 and MMP-9 levels and increasing ZO-1 and Occludin expression.Moreover,its anti-aging mechanism may involve reducing p21 mRNA expression and limiting telomere shor-tening.

外文关键词：

bone marrow stem cellstrokeQixiong Zuogui Compound Prescriptionagingexosomesblood-brain barrier

作者：

要莹莹、樊飞燕、李少康、施艾每、杨俊红、张运克

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作者单位：

河南中医药大学第一临床医学院,河南郑州 450046

河南中医药大学第一附属医院,河南郑州 450000

关键词：

骨髓间充质干细胞脑卒中芪芎左归复方衰老外泌体血脑屏障

基金：

河南省中原英才计划(育才系列)-中原科技创新领军人才项目河南省中医药科学研究专项河南省中医学"双一流"创建科学研究专项

项目编号：

2242005100272023ZY1030HSRP-DFCTCM-2023-1-04

出版年：

2024

DOI：

10.19540/j.cnki.cjcmm.20240524.501

中国中药杂志

中国药学会

中国中药杂志

CSTPCD北大核心

影响因子：1.718

ISSN：1001-5302

年,卷(期)：2024.49(18)