ABSTRACT
Objective Isohemia-reperfusion injury oecurred during heart transplantation may result in failure of grafts and the death of receivers perioperatively. Over expression of TGF-β1 in the myocardium therapeutically was shown to be help-ful in limiting the reperfusion injury to the grafts. The study was designed to investigate the role of Ad. mTGF-β1 gene transfec-tion during ischemia-reperfusion injury in vitro after heart transplantation in rats and the possible mechanisms. Methods The model of heterotopic cardiac transplantation was established by Heron's technique with cuff vessel anastomosis. Animals were divided into 3 groups: in group A ( n =6, control group), the donor hearts were perfusod with 6 ml of Stanford University cardio-plegic solution via coronary arteries at 4℃ for about 40 minutes; in group B ( n =6, vector alone group), the donor hearts were perfused with 6 ml of Stanford University cardioplegic solution containing 5 × 10~9 plaque-forming units( pfu)/gram of the vec-tor, and in group C (study group), the donor hearts were perfused with the solution containing 5 × 10~9 pfu/gram vector with mTGF-β1. The donor hearts were observed with an electro-microscope. The expression of mTGF-β1 in the grafts was identified with immunohistochemical staining. Gene products expressed in tissues were quantified by one step RT-PCR. Activities of SOD ,MDA ,MPO in the grafts were measured. Results At 8 hours after transplantation, mTGF-β1 and its expression were de-tected by means of RT-PCR and immunohistochemical staining in the rats of group C. Expression scores of foreign gene were significantly higher in groups A and B. The apoptotic index of the myocardial cells in group C was lower than those in groups A and B. The activity of SOD was higher in group C than those in groups A and B, though the activities of MDA and MPO were decreased. Conclusion The study demonstrated that gent transfer in vitro via coronary artery was effective. Ad. mTGF-β1 gene transfection in vitro ameliorates the myocardial ischemia-reperfusion injury in rats, for which heart transplantation was per formed, increases the activity of SOD, and decreases the activities of MDA, MPO.
ABSTRACT
BACKGROUND:Autologous peripheral blood hematopoietic stem cell transplantation,as the current latest therapy is widely used in the treatment of autoimmune diseases,however,the treatment on myasthenia gravis is still in the primary stage.OBJECTIVE:To create the model of myasthenia gravis with autologous peripheral blood hematopoietic stem cell transplantation in rats and to observe clinical manifestation,attenuation rate of repetitive nerve electric stimulation action potential and titer of serum acetylcholine receptor Ab in the models.DESIGN,TIME AND SETTING:The randomized control animal experiment was performed at the Human Anatomy Laboratory,Medical College of Zhengzhou University between July and December 2007.MATERIALS:Totally 50 female Wistar rats were randomly assigned into four groups,a normal group(n=10),an adjuvant control group(n=10),a stem cell transplantation group(n=15),and a model control group(n=15).METHODS:Rats in the normal group were intact.Rats in the adjuvant group were subjected to the same volume of saline and cyclophosphamide.Rats in the stem cell transplantation group were used to make experimental autoimmune myasthenia gravis by intraperitoneally infusing with serum of myasthenia gravis patients.Bone marrow stem cells were mobilized by granulocyte colony-stimulating factor.Autologous peripheral blood stem cells were collected and then were transplanted via tail vein injection after the pretreatment regimen of cyclophosphamide.Rats in the model control group were treated with an equal volume of peripheral blood,and with other procedures as the stem cell transplantation group.MAIN OUTCOME MEASURES:Swimming time and survival rate of rats from each group after stem cell transplantation;attenuation rate of repetitive nerve electric stimulation action potential and titer of serum acetylcholine receptor Ab were measured at 4 and 9 weeks after stem cell transplantation.RESULTS:The survival rate was higher in the stem cell transplantation group than in the model control group(P