血管内皮细胞中铁调素对成骨细胞功能的影响  

Effects of hepcidin in vascular endothelial cells on osteoblastic function

作　　者：丁奕栋 王爱飞[1] 张辉[1] 徐又佳[1] Ding Yidong;Wang Aifei;Zhang Hui;Xu Youjia(Department of Orthopedics,the Second Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第二医院骨外科,215000

出　　处：《中华实验外科杂志》2021年第7期1199-1201,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81874018);姑苏卫生人才计划(GSWS2019004)。

摘　　要：目的:探讨血管内皮细胞中铁调素(HAMP)变化对成骨细胞的影响及其作用机制。方法:设计铁调素敲降和过表达的慢病毒,对血管内皮细胞进行转染72 h。实验组分为4组:血管内皮细胞分为铁调素过表达组(HAMP+组)及其对照组(Cont+组),铁调素敲降组(HAMP-组)和其对照组(Cont-组)。间接共培养的成骨细胞分组同血管内皮细胞分组。蛋白质印迹法(Western blot)检测血管内皮细胞铁调素蛋白(HAMP)、外泌体表面标记蛋白肿瘤易感基因101蛋白(TSG101)和CD63、成骨细胞蛋白碱性磷酸酶(ALP),骨钙蛋白(OCN),RUNT相关转录因子2(RUNX2),荧光定量聚合酶链反应(RT-qPCR)检测成骨相关基因的转录组表达。组间比较采用t检验分析。结果:检测转染的血管内皮细胞中,HAMP+组铁调素蛋白表达(1.21±0.02,t=5.80,P<0.05)高于对照Cont+组。HAMP-组铁调素HAMP-组蛋白表达(0.76±0.02,t=6.56,P<0.05)低于对照Cont-组。共培养后成骨细胞成骨蛋白ALP、OCN、RUNX2表达,在HAMP+组成骨细胞中(1.07±0.01、1.61±0.03、1.94±0.01,t=313.40、184.30、309.50,P<0.05)高于Cont+组;在HAMP-组中蛋白表达低于对照Cont-组(0.70±0.05、0.74±0.01、0.75±0.01,t=3.52、15.90、17.67,P<0.05)。结论:血管内皮细胞内铁调素表达量改变会对成骨细胞产生影响,机制可由"铁调素-血管内皮细胞-成骨细胞"途径驱动,提示了铁调素对骨代谢的重要作用。Objective To explore the effect and mechanism of hepcidin(HAMP)changes in vascular endothelial cells(ECs)on osteoblasts at cellular level.Methods The lentivirus of knockdown and overexpressed into ECs for 72 h.The experimental group was divided into four groups:the HAMP overexpression group(HAMP+group)and its control group(Cont+group),the HAMP knockdown group(HAMP-group)and its control group(Cont-group).Osteoblastswere divided as ECs.Western blotting was used to detect the expression of HAMP,exosomal marker proteins tumor susceptibility 101(TSG101)and CD63,osteogenic proteinsalkaline phosphatase(ALP),osteocalcin(OCN)and Runt-related transcription factor 2(RUNX2),and real-time quantitative polymerase chain reaction(RT-qPCR)were used to detect the transcriptome expression of osteogenic genes.T-test was used in statistical analyses.Results The level of HAMP protein in HAMP+group was significantly increased(1.21±0.02,t=5.80,P<0.05),and in HAMP-group was significantly decreased(0.76±0.02,t=6.56,P<0.05).The ECs with overexpression and knockdown of HAMP were successfully constructed.Osteogenic protein in HAMP+group MC3T3 were significantly higher than Cont+group(1.07±0.01,1.61±0.03,1.94±0.01,t=313.40,184.30,309.50,P<0.05),in HAMP-group were significantly lower than Cont-group(0.70±0.05,0.74±0.01,0.75±0.01,t=3.52,15.90,17.67,P<0.05).Conclusion Changes of HAMP expression index in ECs can affect osteogenicfunction,and the mechanism can be driven by HAMP_vascular endothelial cells_osteoblasts,reflecting the indirect effect of HAMP on bone metabolism.

关 键 词：铁调素 成骨细胞 血管内皮细胞 骨代谢 

分 类 号：R68[医药卫生—骨科学]