摘要
目的探讨培哚普利对糖皮质激素(GCs)诱导的骨髓间充质干细胞(MSCs)凋亡及成骨分化的影响,并分析可能的机制。方法将体外培养的MSCs分为5组:空白对照组、地塞米松组(DEX组,10^(6)mol/L DEX)、培哚普利低、中、高剂量组(10^(6) mol/L DEX溶液+0.01μmol/L培哚普利、10^(6)mol/L DEX溶液+0.1μmol/L培哚普利、10^(6) mol/L DEX溶液+1.0μmol/L培哚普利)。运用MTT法、流式细胞术分别检测细胞增殖、凋亡情况;通过实时荧光定量PCR法、免疫印迹法分别检测半胱氨酰天冬氨酸特异性蛋白酶3(cleaved Caspase-3)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关蛋白(Bax)及碱性磷酸酶(ALP)、Ⅰ型胶原(CollagenⅠ)、骨桥蛋白(OPN)以及Notch通路配体(JAG1、JAG2)、跨膜受体蛋白(Notch1、Notch2)、下游靶基因(Hes1、Hey1)mRNA及其蛋白表达情况。结果与空白对照组比较,DEX组MSCs增殖抑制率、凋亡率、Bcl-2、cleaved-caspase-3 mRNA及其蛋白表达升高,ALP、CollagenⅠ、OPN、Bax、JAG1、JAG2、Notch1、Notch2、Hes1、Hey1 mRNA及其蛋白表达降低(P<0.05);与DEX组比较,培哚普利低、中、高剂量组MSCs增殖抑制率、凋亡率、Bcl-2、cleaved-caspase-3 mRNA及其蛋白降低,ALP、CollagenⅠ、OPN、Bax、JAG1、JAG2、Notch1、Notch2、Hes1、Hey1 mRNA及其蛋白表达升高(P<0.05),呈剂量依赖性。结论培哚普利能够抑制DEX诱导的MSCs凋亡,并促进成骨分化,机制可能与促进Notch通路活化有关。
Objective To investigate the effect of perindopril on the apoptosis and osteogenic differentiation of bone marrow mesenchymal stem cells(MSCs)induced by glucocorticoids(GCS),and to analyze the possible mechanism.Methods The MSCs were cultured in vitro and randomly divided into 5 groups:blank control group,dexamethasone group(DEX group,10^(6) mol/L DEX),low,medium and high dose perindopril groups(10^(6) mol/L DEX solution+0.01,0.1,1.0μmol/L perindopril).MTT assay and flow cytometry were used to detect cell proliferation and apoptosis;Real-time fluorescent quantitative PCR,Western blot was used to detect the mRNA and expression of apoptosis related proteins cysteiny aspartate specific protease3(cleaved Caspase-3),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),osteogenic differentiation specific proteins alkaline phosphatase(ALP),typeⅠcollagen(Collagen I),osteopontin(OPN),Notch pathway ligands(JAG1,JAG2),transmembrane receptor proteins(Notch1,Notch2),and downstream target genes(Hes1,hey1).Results Compared with those in the blank control group,the proliferation inhibition rate and apoptosis rate of MSCs,Bcl-2,cleaved-caspase-3 mRNA and protein expression in dex group were significantly increased(P<0.05),the mRNA and protein expression of ALP,Collagen I,OPN,Bax,JAG1,JAG2,Notch1,Notch2,Hes1,Hey1 was significantly decreased(P<0.05);compared with those in DEX group,the proliferation inhibition rate and apoptosis rate of MSCs,Bcl-2,cleaved-caspase-3 mRNA and protein expression in low,medium and high dose perindopril groups were significantly decreased(P<0.05),the mRNA and protein expression of ALP,Collagen I,OPN,Bax,JAG1,JAG2,Notch1,Notch2,Hes1,Hey1 was significantly decreased(P<0.05),in a dose-dependent manner.ConclusionPerindopril can inhibit DEX-induced apoptosis and promote osteogenic differentiation of MSCs.The mechanism may be related to the activation of Notch pathway.
作者
朱治同
李泽平
董立明
ZHU Zhitong;LI Zeping;DONG Liming(Department of Orthopedics, Affiliated Hospital of Zunyi Medical University, Zunyi 563000, China)
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2022年第2期230-236,共7页
Chinese Journal of Osteoporosis
关键词
培哚普利
糖皮质激素
骨髓间充质干细胞
凋亡
骨分化
perindopril
glucocorticoids
bone marrow mesenchymal stem cells
apoptosis
bone differentiation
作者简介
通信作者:董立明,Email:Dongliming777@163.com。
