Effects And Mechanisms Of Rehmannia Glutinosa Oligosaccharide, Nkx2.5Gene Transfected Bone Marrow Mesenchymal Stem Cells Transplantation With Heart Failure

Background: Heart failure is one of the major serious diseases to do harm to humanhealth. Drugs, interventional treatment and heart transplantation are currently themain methods to treat HF. However, they fail to improve the patient’s condition.Stem cells can regenerate functional cardiomyocytes and it is promising to become anew therapeutic measure to treat heart failure.Objective: To study the effects and mechanisms of RGOs and Nkx2.5genetransfected BM-MSCs transplantation with heart failure.Methods: BM-MSCs were isolated and amplified in vitro, then P3generation ofBM-MSCs were infected by lentivirus carrying Nkx2.5gene. When cultured toP5-generation, BM-MSCs labeled with Brdu were used for stem cell transplantation.Heart failure models were duplicated by injecting doxorubicin (total dose of15mg/kgwt) intraperitoneally in male SD rats. When the models were prepared, rats wererandomly divided into5groups: HF、BM-MSCs、Nkx2.5+BM-MSCs、RGOs andCombined group. Echocardiography was used to detect cardiac function in rats.Then the animals were sacrificed. The wet hearts were weighed and calculateheart-to-body-weight ratio (WW/BW). Immunohistochemistry was used to studythe related parameters and HE staining was used to determine the pathologicalchanges. Collagen volume fraction by Masson staining was detected to measuremyocardial fibrosis. The total mRNA were extracted to detect the following genesincluding MEF2、GATA4、 cTnI、CX43、 TGF-β1、CollagenⅠby Q-PCR.Protein of Myocardial tissue were extracted to detect the expression of MEF2、GATA4、 cTnI、 CX43by western blot.Results:1.Rats with heart failure manifest as weight loss and heart systolic、diastolic、pump function decrease.2. Brdu-immunohistochemistry suggested that hearts of BM-MSCs、Nkx2.5+BM-MSCs and Combined group exist cells from stem cells.3.The treatments increased rats’body weight and reduced WW/BW.4. The systolicfunction of heart was improved in all treatment groups, and the Combined group wasbestly inproved. The pump function of heart was improved in all treatment groups,the effect was much more improved in Nkx2.5+BM-MSCs and Combined group thanin BM-MSCs group.5. HE staining showed that the myocardial cells edema,steatosis and necrosis in all treatment groups were extenuated when compared withHF groups, and there were significant Capillary proliferation in all treatment groups.6. Compared with control group, CVF was significantly increased in HF group, CVFin BM-MSCs、Nkx2.5+BM-MSCs、RGOs and Combined group was decreasedcompared with HF group, CVF in Combined group was lower than the other3treatment groups.7. Compared with control group, the expression of TGF-β1andcollagen Ⅰw as higher inHF group. BM-MSCs and RGOs reduced the expressionof TGF-β1and collagen Ⅰi n heart failure, the expression level of TGF-β1andCollagen Ⅰi n Combined groupdecreased mostly when compared with the other twogroups.8. CX43was decreased in HF group, BM-MSCs and RGOs increased theexpression level of CX43, Nkx2.5gene can strengthen the effect of BM-MSCs.9.The expression of MEF2and GATA4was decreased in HF group, MEF2and GATA4was improved in all treatment groups, the increase was much more remarkable inNkx2.5+BM-MSCs and Combined group than in BM-MSCs group.Conclusions:1BM-MSCs、Nkx2.5gene transfected BM-MSCs、RGOs and combined treatment areeffective in improving heart systolic and pump function, decreasing myocardialfibrosis，restraining ventricular remodeling. The effects of Combined treatment ismuch better.2BM-MSCs、Nkx2.5gene transfected BM-MSCs、RGOs and Combined treatmentdecrease myocardial pathological changes, combined treatment is the most evident.BM-MSCs、Nkx2.5gene transfected BM-MSCs、RGOs and combined treatmentimprove myocardial CX43expression, Nkx2.5gene transfected BM-MSCs andcombined treatment is better than other groups. 3BM-MSCs、Nkx2.5gene transfected BM-MSCs、RGOs and combined treatmentdecrease collagen volume in myocardial tissue and reduce the expression TGF-β1andcollagen Ⅰ, combined treatment has the strongest effect.4BM-MSCs、Nkx2.5gene transfected BM-MSCs、RGOs and combined treatmentenhance GATA4, MEF2expression to improve heart function, inhibit ventricularremodeling function.5. Nkx2.5gene transfection enhances the effects of BM-MSCS in improve GATA4,MEF2expression, significantly improve the systolic and pump function.6RGOs can cooperate with Nkx2.5gene transfected BM-MSCs in inhibiting theTGF-β1expression, enhance its efficacy of decreasing myocardial fibrosis andinhibiting ventricular remodeling.