[Myocardial regeneration was promoted by injected Oct4 adenovirus eukaryotic expression vector]

To construct octamer-binding transcription factor-4 (Oct4) adenoviruse eukaryotic expression vector pAV.Ex1d.-CMV>mOCT4/IRES/eGFP, and study its effect on myocardial regeneration in mice.

We constructed eukaryotic expression vector pAV.Ex1d.-CMV>mOCT4/IRES/eGFP using Gateway(R);technology and then packaged the vector with adenoviruses. After adenovirus packaging, the vector was injected into the mouse myocardium by microsyringe. The expression of Oct4 was determined by the methods of immunofluorescence and RT-PCR, and hematoxylin-eosin (HE) staining was used to detect whether the myocardial tissue was changed after the expression of Oct4. We also constructed mouse myocardial infarction model and identified the infarct area using HE staining for injecting the Oct4 adenovirus vector into the area surrounding the infarct area, Four weeks later, we detected the expression of cardiac troponin T in the infact area by Western blotting.

Immunofluorescence showed red fluorescence indicating that Oct4 was expressed in the myocardial cell nucleus, but it could not be detected in the control group and the the control virus group. RT-PCR also proved the expression of Oct4 in Oct4 adenovirus vector group but not in the control virus group and the control group. There was no pathological changes in the myocardial tissues after Oct4 was expressed in vivo. Four weeks after the adenovirus vector was injected around the infarct area, the expression level of cardiac troponin T in Oct4 group was significantly different from that in the control group or the control empty virus group (P<0.5).

Exogenous Oct4 can be expressed in the mature myocardial tissues in vivo and promote myocardial regeneration.