健脾补肾活血方含药血清调控PI3K/Akt信号通路影响大鼠内皮祖细胞功能的机制  

Mechanism of Serum Containing Jianpi Bushen Huoxue Recipe Regulating PI3K/Akt Signaling Pathway and Affecting the Function of Rat Endodermal Progenitor Cells

作　　者：张伟[1] 刘向哲[1] 韩红艳[2] 张燕平[1] 路永坤[1] 周红霞[1] Zhang Wei;Liu Xiangzhe;Han Hongyan;Zhang Yanping;Lu Yongkun;Zhou Hongxia(The First Afiliated Hospital of Henan University of TCM,Zhengzhou 450000,China;Department of Acupuncture-Moxibustion and Tuina of Henan University of TCM,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]河南中医药大学针灸推拿学院,郑州450046

出　　处：《世界科学技术-中医药现代化》2023年第1期227-234,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：河南省科学技术厅重点研发与推广专项(科技攻关)(212102310360):基于关键证候的急性缺血性中风中医药干预方案循证评价研究,负责人:刘向哲;河南省卫生健康委员会中医药科研专项重点课题(2019JDZX2008):急性缺血性中风病证结合序贯治疗方案循证评价及机制研究,负责人:刘向哲;河南省中医管理局中医药科研专项普通课题(2019ZY2031):健脾补肾活血方治疗脑小血管病白质病变MCI临床疗效评价,负责人:刘向哲。

摘　　要：目的探讨健脾补肾活血方含药血清影响大鼠骨髓源内皮祖细胞(EPCs)增殖、迁移、成管功能的机制,以期为该方治疗缺血性脑血管病提供理论依据。方法SD大鼠12只,随机分为两组:中药组和对照组,每组6只。分别给予每毫升含生药为3.25 g健脾补肾活血方颗粒药液和CMC-Na溶液灌胃,采集血清,组内混合,保存备用。SD大鼠2只,取骨髓EPCs,并培养、鉴定。将EPCs随机分为8组:空白组(KB),低、中、高浓度含药血清组(Lx、Mx、Hx),PI3K抑制剂组(PI),低、中、高浓度含药血清+PI3K抑制剂组(Lp、Mp、Hp)。分别给予同等体积的对照组混合血清,10%、20%、40%浓度中药组混合含药血清,30μmol·L^(-1)浓度LY294002,10%、20%、40%浓度中药组混合含药血清+30μmol·L^(-1)浓度LY294002。Western blot、CCK8、Tranwell、体外成血管试剂盒分别检测各组EPCs p-Akt/Akt、p-PI3K/PI3K的相对蛋白表达量、增殖、迁移、成管能力。结果与空白组(KB)比较,健脾补肾活血方低、中、高浓度含药血清组(Lx、Mx、Hx)EPCs p-Akt/Akt、p-PI3K/PI3K的相对蛋白表达量、增殖、迁移、成管能力明显提高(P<0.05)。PI3K抑制剂组(PI)和空白组(KB),低、中、高浓度含药血清+PI3K抑制剂组(Lp、Mp、Hp)和与之分别对应的Lx、Mx、Hx组相比较,EPCs p-Akt/Akt、p-PI3K/PI3K的相对蛋白表达量、增殖、迁移、成管能力明显下降(P<0.05)。结论健脾补肾活血方可调控PI3K/Akt信号通路提高大鼠骨髓源EPCs增殖、迁移及成管能力。Objective To explore the mechanism of Jianpi Bushen Huoxue Recipe containing serum affecting the proliferation,migration and tube forming function of rat bone marrow-derived endothelial progenitor cells(EPCs),in order to provide a theoretical basis for the treatment of ischemic cerebrovascular disease.Methods Twelve SD rats were randomly divided into two groups:Traditional Chinese medicine group and control group,with 6 rats in each group.Give 3.25G Jianpi Bushen Huoxue Decoction granule solution and CMC Na solution per ml of crude medicine respectively,collect serum,mix in the group and keep it for standby.EPCs from bone marrow of 2 SD rats were cultured and identified.EPCs were randomly divided into 8 groups:blank group(KB),low,medium and high concentration drug containing serum group(LX,MX,HX),PI3K inhibitor group(PI),low,medium and high concentration drug containing serum + PI3K inhibitor group(LP,MP,HP).They were given the same volume of mixed serum in the control group and mixed drug containing serum in the traditional Chinese medicine group at the concentration of 10%,20% and 40%,respectively 30 μmol·L^(-1) LY294002,10%,20%,40% concentration of traditional Chinese medicine group mixed drug containing serum + 30 μmol·L^(-1) concentration LY294002.Western blot,CCK8,tranwell and in vitro angiogenesis kit were used to detect the relative protein expression,proliferation,migration and tube forming ability of EPCs p-Akt/Akt and p-PI3K/PI3K in each group.Results Compared with the blank group(KB),the relative protein expression,proliferation,migration and tube forming ability of EPCs p-Akt/Akt and p-PI3K/PI3K in the low,medium and high concentration serum group(LX,MX and HX) of Jianpi Bushen Huoxue recipe were significantly improved(P0.05).The relative protein expression,proliferation,migration and tube forming ability of EPCs p-Akt/Akt and p-PI3K/PI3K decreased significantly in PI3K inhibitor group(PI) and blank group(KB),low,medium and high concentration serum +PI3K inhibitor group(LP,MP,HP) and their corres

关 键 词：健脾补肾活血方 内皮祖细胞(EPCs) PI3K/AKT信号通路 缺血性脑血管病 血管新生 

分 类 号：R285.5[医药卫生—中药学]