人参皂甙Rd通过PI3K/AKT/mTOR/Hif-1α通路促进成年大鼠缺血性脑卒中后的血管发生  

Ginsenoside Rd promotes angiogenesis of rat after cerebral ischemic stroke through PI3K/AKT/mTOR/Hif-1α pathway

作　　者：曽联婷 史明[1] 惠娟[1] 郝明华[1] 瓮调调 韩军良[1] 

机构地区：[1]第四军医大学西京医院神经内科,西安710032

出　　处：《神经解剖学杂志》2015年第6期686-692,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81073094)

摘　　要：目的:探索人参皂甙Rd(Ginsenoside Rd,GSRd)对成年大鼠缺血性脑卒中血管发生的作用及可能机制。方法:80只成年雄性Sprague-Dawley大鼠,体重280~300 g,用线栓法建立局灶性短暂性大脑中动脉阻塞模型(focal transient middle cerebral artery occlusion,MCAO),Sham为手术但不阻塞。随机分为四组,每组20只:Sham+生理盐水(SA)、Sham+GSRd、MCAO+SA、MCAO+GSRd。术后3 d,通过RECA-1免疫荧光组织化学染色标记血管内皮细胞、并计算梗死灶周围微血管的密度和分支点。通过Western Blot检测梗死侧皮层血管内皮细胞生长因子(vascular endothelial growth factor,VEGF),低氧诱导因子(hypoxia-inducibal factor 1 alpha,Hif-1α),磷酸化的哺乳动物雷帕霉素靶蛋白(p-mammalian target of rapamycin,p-mTOR),磷酸化的蛋白激酶B(p-protein kinase B,pAKT)的蛋白表达水平。结果:(1)免疫荧光组织化学染色结果显示:与MCAO+SA对照组相比,MCAO+GSRd治疗组的血管数(518.75±34.88/mm^2 vs 391.40±17.66/mm^2,P<0.001)和分支点(255.47±36.05/mm^2 vs 203.13±12.05/mm^2,P<0.01)显著增多。Sham+SA和Sham+GSRd的血管数(503.12±45.32/mm^2 vs 492.18±61.93/mm^2)和分支点(270.31±35.94/mm^2 vs 258.59±42.25/mm^2)则无显著差异;(2)Western Blot结果显示:与对照组相比,GSRd显著增加VEGF,Hif-1α,p-mTOR,p-AKT的蛋白表达水平(P<0.05)。(3)mTOR特异性阻断剂雷帕霉素抑制了GSRd对Hif-1α(P<0.01)、VEGF(P<0.05)的作用。结论:人参皂甙Rd可促进成年大鼠脑卒中后的血管发生,其对血管发生的作用可能是通过PI3K/AKT/mTOR/Hif-1α通路所介导。Objective：To investigate the effect of Ginsenoside Rd（GSRd） on angiogenesis after cerebral ischemic stroke in adult rats and underlying potential mechanisms.Methods：Eighty adult male Sprague-Dawlay rats,weighting280-300 g,subjected to focal transient middle cerebral artery occlusion（MCAO）,were randomly divided into four groups,20 rats in every group：Sham ＋ saline（SA）;Sham ＋ GSRd;MCAO ＋ SA;MCAO ＋ GSRd.Post stroke day 3,RECA-1 was used to marking vascular endothelial cells in penumbra of ipsilateral cortex through immunofluorescence histochemical staining.And Western Blot was used to detect the expression changes of vascular endothelial growth factor（VEGF）,hypoxia-inducibal factor 1 alpha（Hif-1α）,p-mammalian target of rapamycin（p-mTOR）,p-protein kinase B（p-AKT） levels.Results：（1） Immunofluorescence histochemical staining showed that compared with MCAO ＋ SA control group,microvessel density（391.40 ± 17.66/mm^2 vs 518.75 ± 34.88 /mm^2,P 0.001） and branch point（203.13 ± 12.05/mm^2 vs 255.47 ± 36.05/mm^2,P〈 0.01） in penumbra area of MCAO ＋ GSRd treatment group were significantly increased.Microvessel density and branch point in Sham ＋ SA and Sham ＋ GSRd were 503.12 ±45.32/mm^2 vs 492.18 ±61.93/mm^2;270.31 ± 35.94/mm^2 vs 258.59 ±42.25/mm^2,respectively.Two groups had no significant differences.（2） Western Blot results showed that compared with control group,GSRd treatment group significantly increased the expressing of VEGF,Hif-1α,p-mTOR,p-AKT protein levels（P〈 0.05）.（3） Rapamycin,the specific inhibitor of mTOR,supressed the effect of GSRd to Hif-1α（P〈 0.01） and VEGF（P〈 0.05）.Conclusion：Conclusion：GSRd could promote angiogenesis of adult male rats after ischemic stroke,the potential mechanism could be PI3K/AKT/mTOR/Hif-1α.

关 键 词：人参皂甙RD 脑缺血 血管再生 VEGF HIF-1Α mTOR AKT 大鼠 

分 类 号：R285.5[医药卫生—中药学]