The Research Of HIF-1α Promoting DPSCs The Differentiation Of Angiogenesis In Vitro

Objective The HIF-1α gene was transfected stably into dental pulp stem cells(DPSCs). Then tissue engineering technique was used to explore the function of HIF-1α gene in promoting the differentiation of DPSCs into blood vessels in vitro.Methods HIF-1α gene was mutated(402, 564,and 803 point)to maintain stable expression and high activity under normoxic conditions in vitro. The lentiviral vectors of Lenti-MT, Lenti-WT and Lenti-Lac Z 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 m RNA and protein were detected. The influence on cell proliferation from lentiviral vectors was detected by MTT. After transduction, cells were collected on days 0, 1, 4, 7, 14 and 21, RNA and protein were extracted, the expression of angiogenic factor was detected by q PCR and Western blot to compare these three lentiviral vectors’ function on DPSCs.Results DPSCs were transfected virus,the MOI value was set at 10 pfu / cell. After 7 days, we observed the viral transfection efficiency was best(90%) by inverted fluorescence microscopy. MTT showed that absorbance values of Lenti-WT and Lenti-MT group were greater than Lenti-Lac Z group and MT group was greater than WT group. DPSCs proliferation was promoted by HIF-1α gene. The results of q PCR and Western blot showed that HIF-1α gene was located in cell nucleus, expression of target gene was detected at both m RNA and protein level. After gene transduction, cells were collected on days 0, 1, 4, 7, 14 and 21, RNA and protein were extracted, theexpressions of angiogenic factors(VEGF、SDF-1 and Ang-2) were detected by q PCR and Western blot, expressions of angiogenic factors at the m RNA and protein levels were significantly increased(P<0.05). The formation of blood vessel in Lenti-MT and Lenti-WT groups was better than that in Lenti-Lac Z group(P<0.05), and the best result was found in Lenti-MT group(P<0.05).Conclusion This study deteced and proved that HIF-1α gene could induce DPSCs to express more angiogenic factors(VEGF、SDF-1 and Ang-2) at the m RNA and protein levels in vitro. The result preliminarily confirmed the HIF-1α gene could promote DPSCs to form blood vessels.