Effect Of Pericardial Injection Of Adipose-derived Stem Cells On Heart Failure In Rats

BackgroundAt different stages of heart disease development,stem cells can reach their target organs in various ways.Intrapericardial injection of ADSCs in rats with heart failure has not been studied,and the safety and practicality of intrapericardial injection of ADSCs in rats with heart failure have not been reported.ObjectiveTo investigate the therapeutic effect of ADSCs injecting through the pericardial cavity on the heart of rats with heart failure,and to provide support for basic research and practical fields of stem cell therapy in heart failure.Methods1.SD rats(young adults with better cell activity)at 4-6 weeks were selected,the apparatus was sterilized at high temperature,and adipose tissue is taken from the inguinal skin under aseptic conditions for the separation and culture of ADSCs.The dryness was identified by IHC staining and IF staining,and the selected surface markers were CD29,CD44,CD73,and CD45.2.Male SD rats,6-8 weeks old,were selected as models and divided into 3 form including HF,treatment(T group)and a control group(C group)without surgery.The HF model was created by injecting drug(doxorubicin hydrochloride)in 15 SD rats in the T group and the HF group,10 rats in the C group were injection saline in the same way.There were 40 animals in three groups.and the heart function of SD rats in the three groups were detected by ultrasound for small animals,respectively.LVEF<50%was considered to be successful,and the mortality rate of this model was about 1/3.3.The identified ADSCs were injected into the treated rats through the pericardial cavity.In living imaging and histological sections were performed to see if the cells entered the heart through the endocardium.4.The proteins were extracted from fresh rat heart tissue and the hearts were frozen into sections.BNP,α-SMA and CRP were detected by Western-blotting.The same indexes were verified by immunohistochemical staining.Results1.IHC showed that ADSCs expressed these 3 indicators including CD29,CD73 and CD44 except CD45.The single IF results were same with the IHC results,and the double IF results showed that CD44 and CD73 were co-expressed in ADSCs.2.Frozed-section microscopic observation showed that the injected cells were densely distributed under the epicardium 4 days after injection,and scattered in the myocardial tissue 5 days later.In living imaging of small animals showed that cells injected pericardially were full field distributed in the heart,and their fluorescence intensity remained high for 10 days,and gradually decreased after 10 days.3.WB results indicate that only a small amount of BNP was found in the rats in the C group,and a lot of BNP was found in the rats in the HF group,while the BNP content in the T group was decreased.Alpha-SMA levels was approximately the same as that of BNP and was also relatively low in the T group.CRP levels were lower in the C group,slightly more in the HF group,and higher in the T group.IHC results were same with WB results.ConclusionsThe injection of adipose-derived stem cells through pericardial cavity can penetrate epicardial membrane,migrate into myocardium,and decrease some indexes of heart failure,have certain therapeutic effect on heart failure.