TAZ激动剂TM-25659促进人脂肪干细胞成骨的体内外研究  

The TAZ chemical activator TM-25659 facilitates osteogenic differentiation of human ADSCs in vitro and in vivo

作　　者：胡佳安 吴亚平[1] 黄蓉[1] 杨建荣[2] 王琰玲[1] 朱玉敏[1] Hu Jia’an;Wu Yaping;Huang Rong;Yang Jianrong;Wang Yanling;Zhu Yumin(Jiangsu Key Laboratory of Oral Diseases,Nanjing Medical University,Nanjing 210029;Department of Oral and Maxillofacial Surgery,the Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029 China)

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [2]南京医科大学口腔医学研究所,南京医科大学附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2019年第7期971-977,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京医科大学科技发展基金(2016NJMU058,08NMUM059

摘　　要：目的:探索成骨转录共刺激因子TAZ(Tafazzin)激动剂TM-25659对人脂肪干细胞(adipose-derived stem cells,ADSCs)在体内外成骨分化和骨形成过程中的生物学作用。方法:分离培养人ADSCs,通过蛋白质免疫印迹、实时定量PCR检测ADSCs成骨、成脂分化及TM-25659干预后不同时间点TAZ的表达变化,茜素红和油红O染色分别验证其成骨和成脂效果;以β-磷酸三钙(β-tricalcium phosphate,β-TCP)材料为载体负载成骨诱导和TM-25659预处理后的ADSCs植入裸鼠体内,6周后利用苏木精-伊红染色、Masson三色染色和免疫组织化学染色法检测比较ADSCs体内成骨效果。结果:人ADSCs体外成骨分化过程中TAZ表达上调,而在成脂分化中其表达降低。TM-25659促进ADSCs中TAZ的表达,并增强ADSCs体内成骨及骨生成能力。结论:TAZ是促进ADSCs成骨分化的关键调控因子之一,以TAZ为药物靶点可促进ADSCs的骨再生及修复。Objective:The present study aims to determine whether pharmacological activation of the transcriptional coactivator with PDZ-binding motif Tafazzin(TAZ)by chemical TM-25659 can promote osteogenic differentiation and bone formation of adiposederived stem cells(ADSCs)in vitro and in vivo. Methods:Human ADSCs were isolated and expanded in vitro. The changing expressions of TAZ during ADSCs differentiation at different time points were measured by Western blot and real-time quantitative reverse transcription polymerase chain reaction(PCR). Alizarin red S and oil red O staining were used to detect osteogenic and adipogenic effects. The ADSCs following initial osteogenic differentiation and TM-25659 were loaded on scaffold β-tricalcium phosphate(β-TCP)and then subcutaneously implanted into nude mice. Six weeks later,the results of bone formation in vivo after implantation of the ADSCs were measured and compared by Hematoxylin-Eosin staining,Masson staining and immunohistochemistry staining. Results:TAZ was upregulated during osteogenic differentiation of ADSCs but downregulated during adipogenic differentiation. Pharmacological activation of TAZ by TM-25659 promotes osteogenic differentiation of ADSCs in vitro. The transient treatment of ADSCs with TM-25659 significantly enhances new bone regeneration of ADSCs in vivo. Conclusion:TAZ is a key mediator for promoting osteogenic differentiation of ADSCs. The pharmacological activation of TAZ in ADSCs might become a feasible treatment for bone regeneration and repair.

关 键 词：脂肪干细胞 成骨 Hippo通路 TM-25659 TAZ 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]