首页|基于玄府开阖研究葛根、石菖蒲对三七总皂苷改善脑缺血再灌注模型大鼠血脑屏障损伤的增效作用研究

基于玄府开阖研究葛根、石菖蒲对三七总皂苷改善脑缺血再灌注模型大鼠血脑屏障损伤的增效作用研究

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目的 观察风药(葛根、石菖蒲)与三七总皂苷配伍后对脑缺血再灌注大鼠的神经保护作用,阐明"风药增效"的作用机制.方法 将140只雄性SD大鼠按随机数字表法分为假手术组、模型组、三七总皂苷组、葛根组、石菖蒲组、葛根+三七总皂苷组、石菖蒲+三七总皂苷组,每组20只.除假手术组外,其余各组采用改良Longa线栓法建立脑缺血再灌注模型.各组给予相应药物干预7 d后,采用Zea-longa评分法进行神经功能评分,TTC染色法测定脑梗死体积,伊文思蓝(EB)染色检测缺血侧脑组织血脑屏障(BBB)通透性,透射电子显微镜观察BBB超微结构,免疫组化染色检测脑组织密封蛋白(Claudin 5)表达,Western blot检测闭锁小带蛋白1(ZO-1)、主要促进因子超家族成员2a(Mfsd2a)、闭合蛋白(Occludin)和P糖蛋白(P-gp)、单羧酸转运蛋白1(MCT1)、乳腺癌抗性蛋白(BCRP)表达.结果 与模型组比较,各给药组大鼠神经功能评分降低(P<0.05),脑梗死率降低(P<0.05),脑组织EB含量下降(P<0.05),脑组织Claudin 5、ZO-1、Mfsd2a、Occludin蛋白表达升高(P<0.05),P-gp、BCRP、MCT1蛋白表达降低(P<0.05),且葛根+三七总皂苷组、石菖蒲+三七总皂苷组Claudin 5、ZO-1、Mfsd2a、Occludin蛋白表达高于各单独用药组(P<0.05),MCT1蛋白表达低于各单独给药组(P<0.05);葛根+三七总皂苷组ZO-1蛋白表达水平高于单独给药组(P<0.05);石菖蒲+三七总皂苷组P-gp蛋白表达低于三七总皂苷组、石菖蒲组及葛根+三七总皂苷组(P<0.05).结论 风药(葛根、石菖蒲)可增强三七总皂苷对脑缺血再灌注大鼠的神经保护作用,其机制可能与保护BBB结构完整性、维持中枢屏障特性,同时调节物质转运蛋白、提高药物脑内含量有关.
Study on the potentiation effect of Puerariae Lobatue Radix and Acori Tatarinowii Rhizoma on PNS in the improvement of blood-brain barrier damage in MCAO rats based on the opening and closing of sweat pores
Objective To observe the neuroprotective effect of wind medicine(Puerariae Lobatue Radix and Acori Tatarinowii Rhizoma)in combination with Panax ginseng total saponin(PNS)on cerebral ischemia-reperfusion rats;To elucidate the mechanism of"wind medicine increasing effect".Methods Totally 140 male SD rats were divided into sham-operation group,model group,PNS group,Puerariae Lobatue Radix group,Acori Tatarinowii Rhizoma group,Puerariae Lobatue Radix+PNS group,Acori Tatarinowii Rhizoma+PNS group according to the random number table method,with 20 rats in each group.Except for the sham-operation group,the cerebral ischemia/reperfusion rat model was established using the modified Longa line bolus method in the remaining groups.After 7 d of administration of the appropriate pharmacologic intervention in each group,neurological dysfunction was evaluated by Zea-longa score after final administration,cerebral infarct volume was determined by TTC staining;blood brain barrier(BBB)permeability of brain tissue on the ischemic side was detected by Evans blue content;BBB ultrastructure of each group of rats was observed by transmission electron microscopy;Claudin 5 protein expression level was detected by immunohistochemistry;Zonula occludens-1(ZO-1),major facilitator supeffamily domain-containing protein 2a(Mfsd2a),Occludin,P-glycoprotein(P-gp),Monocarboxylate Transporters-1(MCT1)and breast cancer resistance protein(BCRP)protein expression levels were detected by Western-blot.Results Compared with the model group,the rat neurological function scores were reduced in each administration group(P<0.05),infarct volume was reduced(P<0.05),EB content of brain tissue decreased(P<0.05),protein expressions of Claudin 5,ZO-1,Mfsd2a and Occludin in brain tissue were elevated(P<0.05),the protein expressions of P-gp,BCRP and MCT1 were reduced(P<0.05),and the protein expressions of Claudin 5,Mfsd2a,and Occludin was higher in the Puerariae Lobatue Radix+PNS group and Acori Tatarinowii Rhizoma+PNS group than that of each group of medication alone(P<0.05),and the protein expression of MCT1 was lower than that of each group of medication alone(P<0.05);the protein expression level of ZO-1 in the Puerariae Lobatue Radix+PNS group was higher than that of the group of medication alone(P<0.05);P-gp protein expression was lower in Acori Tatarinowii Rhizoma+PNS group than in the PNS group and Acori Tatarinowii Rhizoma group(P<0.05).Conclusion Wind medicine(Puerariae Lobatue Radix and Acori Tatarinowii Rhizoma)may potentiate the neuroprotective effect of PNS on cerebral ischemia-reperfusion rats,and the mechanism may be related to the protection of BBB structural integrity and maintenance of central barrier properties,while regulating substance transport proteins and increasing the intracerebral content of the drug.

Brain ischemiaReperfusion injuryBlood-brain barrierSweat poreWind drugs(TCD)NotoginsenosideRats

郝琳瑶、李双阳、任俊豪、王小强、王凌雪、张德绸、唐红梅、白雪

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西南医科大学中西医结合学院,泸州 646000

西南医科大学附属中医医院神经内科,泸州 646000

西南医科大学中西医结合研究所,泸州 646000

四川省中西医结合医院脑病科,成都 610000

西南医科大学附属中医医院中医药表型组学研究中心,泸州 646000

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脑缺血 再灌注损伤 血脑屏障 玄府 风药(中药) 三七皂苷 大鼠

国家中医药管理局中医药创新团队及人才支持计划项目四川省科技计划四川省中医药管理局科研项目四川省中医药管理局科研项目泸州市科技局重点研发项目西南医科大学附属中医医院学科创新团队

ZYYCXTD-C-2022072022YFS06132022CP17172023MS2762020-SYF-302022-CXTD-05

2024

国际中医中药杂志
中华医学会,中国中医科学院中医药信息研究所

国际中医中药杂志

CSTPCD
影响因子:0.411
ISSN:1673-4246
年,卷(期):2024.46(8)
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