HIF-1α基因介导的牙髓干细胞在体外的成血管作用  

On HIF-1α gene in promoting the formation of blood vessel by DPSCs in vitro

作　　者：邓立方 朱友明[1] 王银龙[1] 

机构地区：[1]安徽医科大学口腔医学院,安徽医科大学附属口腔医院,安徽省口腔疾病研究中心实验室,合肥230032

出　　处：《安徽医科大学学报》2016年第8期1120-1124,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:31501103)

摘　　要：目的探索低氧诱导因子-1α(HIF-1α)基因诱导牙髓干细胞(DPSCs)在体外的成血管作用。方法对HIF-1α进行基因突变,构建突变型、野生型以及对照组的慢病毒载体;体外培养DPSCs;分别用3种慢病毒转染DPSCs后,检测细胞转染效率、目的基因HIF-1α在mRNA及蛋白水平的表达,MTT法检测慢病毒载体对细胞增殖的影响;目的基因转染成功后,q PCR、Western blot法检测HIF-1α调控DPSCs成血管因子的表达。结果 MTT结果表明慢病毒载体对DPSCs的增殖几乎无影响。q PCR和Western blot法检测目的基因HIF-1α成功表达,HIF-1α能够显著上调DPSCs的成血管因子的表达(P<0.05)。突变组和野生组的成血管作用明显强于对照组(P<0.05),而突变组又优于野生组(P<0.05)。结论 HIF-1α基因可以促进DPSCs血管向分化作用。Objective To explore the function of HIF-1α gene in promoting the differentiation of dental pulp stem cells（DPSCs） into blood vessels in vitro. Methods HIF-1α gene was mutated. The lentiviral vectors of Lenti- WT, Lenti-MT and Lenti-LacZ were constructed. The DPSCs were cultured and treated with three lentiviral vectors. Then the cells transduction efficiency and the expression of target gene at the level of mRNA and protein were detected. The influence on cell proliferation from lentiviral vectors was detected by MTT. After transduction, RNA and protein were extracted, the expression of angiogenic factor was detected by qPCR and Western blot. Results MTT showed that there was no effect on DPSCs proliferation from lentiviral vectors. The results of qPCR and Western blot showed that HIF-1α gene was located in cell nucleus, expression of target gene was detected at both mRNA and protein level. After gene transduction, expression of angiogenic factor at the mRNA and protein levels was significantly increased （P 〈 0. 05 ）. The formation of blood vessel in Lenti-MT and Lenti-WT groups was better than that in Lenti-LacZ group （ P 〈 0. 05 ）, and the best result was found in Lenti-MT group （ P 〈 0. 05 ）. Conclusion HIF-1α could promote DPSCs to form blood vessels.

关 键 词：HIF-1Α DPSCs 慢病毒转染 成血管 

分 类 号：R782[医药卫生—口腔医学]