摘要
目的:针对不同胚源的大鼠骨髓间充质干细胞(BMSCs)表现出的衰老差异,探讨组蛋白去甲基化酶Jmjd3对衰老的调控作用。方法:取大鼠下颌骨(M)和胫骨(T)后提取BMSCs原代培养并进行传代,通过β-半乳糖苷酶(SA-β-gal)衰老染色检测细胞衰老特征,茜素红染色检测细胞成骨特征。通过实时定量RT-PCR、Western blot检测Jmjd3及衰老因子在两种BMSCs中的表达。构建sh Jmjd3病毒敲减Jmjd3,转染T-BMSCs检测衰老指标。在大鼠颌骨区域建立骨缺损模型,移植sh Jmjd3修饰的T-BMSCs,通过Micro-CT、Masson染色检测Jmjd3被抑制的T-BMSCs对骨缺损区的修复作用。结果:经SA-β-gal衰老染色,T-BMSCs中衰老细胞数量明显多于M-BMSCs。Jmjd3及衰老因子在M-BMSCs低表达,在T-BMSCs高表达。转染sh Jmjd3病毒导致T-BMSCs中Jmjd3及衰老因子低表达。移植敲减Jmjd3的T-BMSCs,Micro-CT显示其骨平均密度,骨体积分数明显升高,Masson染色显示其骨小梁结构增多。结论:不同胚源大鼠BMSCs的衰老特征具有差异性,将T-BMSCs中Jmjd3抑制以后,可以增强细胞的抗衰老能力,有利于体内的骨修复的治疗。
Objective： To investigate the effects of histone demethylase Jumonji D3 （Jmjd3） on the senescence of rat bone marrow stromal cells （BMSCs） from different embryonic origins. Methods： BMSCs were extracted from mandibular （M） and tibia （T） of rats and cultured for the following experiments. The characteristics of cell senescence were detected by SA-β-gal staining, and osteogenic differentiation were detected by alizarin red staining. The expression of Jmjd3 and senescence related factors, p16 and p21,were detected in M and T group by qRT-PCR and Western Blot. Then, shJmjd3 Lentivirus was constructed and transfected into T-BMSCs to knock down Jmjd3 expression. A critical-sized bone defect model was established in the rat jaw region and shJmjd3-modified BMSCs were transplanted into the defect area. The bone healing region by BMSCs transplantation was detected by Micro-CT and Masson staining. Results： SA-β-gal staining revealed that the number of senescent cells in T-BMSCs was significantly higher than that of M-BMSCs, as well as the expression of Jmjd3 and senescence related factors. T-BMSCs transfected with shJmjd3 resulted in decreased SA-β-gal positive staining cells and declined expression of Jmjd3 and senescence factors. Compared with GFP T-BMSCs, rats transplanted with shJmjd3 T-BMSCs exhibited increased mean bone density and bone volume fraction by Micro-CT, as well as elevated trabecular bone structure by Masson staining. Conclusions： M-BMSCs exhibited lower senescence characteristics than that of T-BMSCs, which have a higher Jmjd3 expression. Inhibition of Jmjd3 in BMSCs can enhance the anti-senescence ability of cells and promote bone repair in vivo.
作者
徐娟
林佳琳
张平
徐荣耀
江宏兵
XU Juan;LIN Jialin;ZHANG Ping;XU Rongyao;JIANG Hongbing(Jiangsu Key Laboratory of Oral Diseases,Department of Oral and Maxillofacial Surgery,Affiliated Stomatological Hospital,Nanjing Medical University,Nanjing 210029,China;Maigaoqiao Community Health Service Center,Nanjing 218000,China)
出处
《口腔生物医学》
2018年第3期115-119,共5页
Oral Biomedicine
基金
国家自然科学基金(81070810)
江苏高校优势学科建设工程资助项目(2014-37)
