Research About Expression Efficacy Of Human Insulin Gene In Type 1 Diabetic Rats Enhanced By Medicine And The Way Plasmids Perfused

ObjectiveExplored again and again, gene therapy technology has become such a pro-mosing method. After so many years' development, gene treatment for di-abebtes mellitus has made much progress, and reached success in a series of animal experiments. But at the same time we face the problom of efficacy and this show us that there is some disdance between experiment and clinical application. The shortness of efficacy mainly include: at present retroviruses are often selected as vectors in the gene therapy. They can carry foreign gene with high molecular weight, integrate into host gene and sustain transgene expression permanently. But the vectors'transfer efficiency is low. They also have low integration efficiency because only the host cells are in dividing foreign gene can integrate into host gene;The proinsulin gene has been mutated in two spots, and so the active insulin can be secreted by non - β cell, but in animal experiment the level of plasma insulin is often low. To enhance the efficacy of gene therapy for diabebtes mellitus and remvove the obstacles for the clinical application, this experiment explores different medicines and the way plasmids infused into animals.MethodsType 1 diabetes mellitus model of rat is estsblished by STZ. After that, we inject HGF and T₃ to the experimental groups ,but don't take any measures to the control group. After the plasmids are perfused in two ways we observe thelevel of blood glucose and plasma insulin. Three months later we kill the rats, extract the rats' liver and make RT - PCR. By comparing expression efficacy of goal gene in each group, we select the best factor that can maximize the expression efficacy.ResultThe group treated with the reorganization plasmids including goal gene which were perfused into the abdominal cavity and HGF and T3 injected 24 hours ago have the best therapy result. The group given HGF and T3 and reorganization plasmids including goal gene which were poured into tail vein have the better therapy result. The group treated with only the reorganization plasmids including goal gene which were perfused into the abdominal cavity have slight effect. The group given only HGF and T3 and the group with no treatment have not any effect. Finally we make RT - PCR and find that the goal gene in experimental group has obvious expression.ConclusionIn animal experiment, through observing the level of insulin and glucose and the changes of weight we can draw the conclusion that perfusing reorganization plasmids including goal gene into the abdominal cavity with injecting HGF and T3 24 hours ago can obviously enhance the transfer and integration efficiency. So this experiment lays the groudwork for the clinical experiment.