SHH信号通路激活对大鼠心脏微血管内皮细胞缺血再灌注损伤血管再生作用研究

[DOI)10.3969/j.issv.1277-0454.6407.27.207心血管疾病专题SHH信号通路激活对大鼠心脏微血管内皮细胞

缺血再灌注损伤血管再生作用研究

国伟，杨军，任法新，梁平平

基金项目：山东省医药卫生科技发展计划项目(2218WSA0615);济宁市第一人民医院博士科研启动基金

作者单位:272211济宁市第一人民医院老年医学科(国伟/;204007青岛大学附属烟台毓璜顶医院(杨军、任法新、梁平平)

通信作者：杨军，E-mail:yuaweil9811227@I/,com

【摘要】目的探讨SHH信号通路激活对大鼠心脏微血管内皮细胞(CMEC)缺血再灌注损伤后促血管再生

的作潜在机制。方法2016年7月一2017年2月大实验。原养SD雄

大鼠CMECs,建立氧/复氧(OGD/R))外缺血再灌注，给予外源SHH蛋白、CyTopaminc(CYC)处理，应用MTT法检测CMECs细胞活力，应用ELISA和RT-TCR方法检测血管新生因子(VEGF、

FGF.Any-I)水平和mRNA表达水平；应用WesWro-Olct方法分别检测SHH信号通路靶分子J SHH、SMO、PychedT、

GU-7GU-0)蛋白表达水平。结果(、)与C组比较，正常氧糖条件下C+SH H组活力增加(P<0.25),但C+CYC组

消减(P<0.25)；当细胞在OGD/R条件下,OGD组CMECs活力与C组比较减弱(P<0.25),而OGD+SH H组与OGD

组比较细胞活力增加(P<0.25)；但OGD+CYC显著消减(戶<0.05)。(2)与C组比较，正常氧糖条件下C+SH H组

血管新生因子VEGF、FGF、Any-I水平和mRNA表达增加(P<0.27),旦C+CYC组上述指标水平和mRNA表达下降

(P<0.01/与C组比较，OGD组VEGF、FGF、Any-I血清水平和mRNA表达显著降低J P<0.01)；OGD+SHH组

VEGF.FGF.AnyA血清水平和mRNA表达显著增加(P<0.07),OGD+CYC组上述指标水平和mRNA表达下降(P<

0.05)。(3)与OGD组比较,OGD+SHH组在OGD/R条件下,SHH信号通路中靶分子SHH、SMO、PychedA、GUA和

GUO蛋白水平明显升高，差义(P<0.25)，但OGD+CYC组SHH信号通路靶分子蛋白表达水平下

降(P<0.27)o结论SHH信号通路激活可增强细胞活力，上调血管新生因子VEGF.FGF和Any-I的表达，在缺血再

灌注损伤后血管再生过程中发作用。

【关键词】心脏微血管内皮细胞;缺血再灌注损伤;血管再生;S HH信号通路;细胞活力;大鼠

【中图分类号】R543.9[文献标识码】A

The sthdy of SHH sivnal pathway activation on raf00^111micravascylae endothenai celUs ischemic repei'fusion injury angiooenesit Guo Wet-,Yang Jun,Ren Faxin,Liang PingpUg.*Depa/mek/g Grontology；JUing First Peopis HospU

toi,ShmUong Province,Aming070217,ChUo

CorrespondUg amOoe:Yang Jun,E-mail:pmoal9811207@

Funding program:SSandopg Province MePicai and HealtO Science and Technology Development Plan Projeei

(2018WS408015)；JUing First Peoples Hospitoi Doctorol Research Stardun Funi

[Abstract i Objectiva To explore the effect of SHH signal pathway activation on the promotion of anyioyenesis aftco ischemipAepeWusion injuo of rat cardiac micovasch/o endothelial cells(CMECs)and its potenCai mechanism.Methodt

Fom July2216to Fedoao2217,the experiment was carried out in Yantai YuPuanydiny Hospital afiliawd to Qinydao Uni­

versity.Primao ch/ure of CMECs in male SD rats was esmUPsheb,oxyyen ydcose dephvatWn/ooxyaenation(OGD/R)sim-

u/wd in vitro ischemipAepeWusion injuo mobei was esmUPshed,exoyenous recombinant human SHH protein and Cyclopam-

mc(CYC)were treated,and CMECs were deteewd by MTT methob Cel l viahidty,usiny ELISA and RT-TCR methobs to dc-

tect the levels of anyioyenesis factors(VEGF,FGF,Any-I/and mRNA expression levels.Westero-O/t method was used to

deWc-the protein expression levels of SHH signaUny pathway taract molechles(SHH,SMO,Pawhed-I,GP-I,GPO).Re-

syltf(7)Compared with youp C,the viahiPty of C+SHH youp increased irnder normoxie ylucose conhiPons(P<0.45),

but the C+CYC aoup decreased(P<0.05)；when cells were undcr OGD/R conhiPons,the viahiPty of CMECs in OGD

doup Compared with youp C,it was weahened(P<0.45),while the cell viahiPty of OGD+SHH youp was increased com­

pared with OGD aoup(P<0.45)；but OGD+CYC was significantly reduced(P<0.45).(0)Compared with youp C,the