The Effect Of Insulin-Producing Cells Which Differentiated From Bone Marrow Stem Cell In Diabetes Mellitus Rat

ObjectiveIn developed countries, Diabetes mellitus DM has changed to be the third non-infectious disease after angiocardiopathy and tumor, it brings heavy burden to the society and economy, and a serious world public healthy problem which became a threaten to human healthy .The research on the differentiation of stem cell into insulin-producing cell provides a new prospect of a cure for DM. In this field, bone marrow stem cell may be one promising kind of stem cells. .Bone marrow stem cell have various differentiation potency and conspicuous plasticity .Many vitro studies have successfuly induced the bone marrow stem cell into insulin-producing cell, which could reverse hyperglycaemia after transplanted into the body of diadet rat, but exvivo transplantion of stem cell has problem of contamination and mutation ,so we investigate whether Gastrin and Epidermal growth factor EGF could induce bone marrow stem cell to insulin-producing cell in vivo and whether the insulin-producing cell that induced by Gastrin and EGF have curativeeffect on diabetic rat.Material and methods40 wistar rats were randomly allocated into four groups: normal group, DM group, mobilization group, mobilization plus induction group, (10 wistar rats in each group). The last three group were treated with single intraperitoneal injection of 2% streptozotocin (STZ, 55 mg/kg body wt) to establish diabetes model. In mobilization group the rats were treated for 5 days with once-daily subcutaneously injected of recombination human Granulocyte-colony stimulating factor (rhG-CSF, 10 mg/kg body wt). The group of mobilization plus induction were treated for 5 days with once-daily subcutaneously injection of rhG-CSF(10 mg/kg body wt), in addition, on the 2nd day after mobilization injection, treat the rats with once-daily intraperitoneal injection of 3ug/ml Gastrin(3ug/kg body wt) and 3ug/ml EGF (1 ug/kg body wt) for continuous 14 days. The groups of normal and DM were treated with correspond dose of NaCl.In the process of experiment, the weight were measured every week,measured the blood glucose every two weeks before treatment and the end of the study measured the fasting serum insulin concentration at 10 weeks, at the end of the study, treat the rats with intraperitoneal injection of 10% Chloral Hydrate (3 ml/kg body wt) to anesthetize them, remove the liver, spleen, pancreas, and then measure the expression of insulin positive cell by immunohistochemistry , the area of insulin positive cell in pancreatic islet were measured by using image analysis software.ResultsBefore administration, the blood glucose and fasting serum insulin concentration level has no significant difference among DM group, mobilization group and mobilization plus induction group. At the end of administration, the blood glucose level of mobilization group and mobilization plus induction group is higher than the normal group but lower that the DM group(p<0.05).At the end of study ,the blood glucose level of mobilization plus induction group is significant lower than DM group (p<0.01),but still higher than normal group , the fasting serum insulin concentration of mobilization plus induction group is significant higher than DM group (p<0.01), compared with normal group, the fasting serum insulin concentration level has no significant difference . Compared with DM group, the blood glucose and fasting serum insulin concentration of mobilization group have no significant difference. In the whole process, the weight of normal group is always higher than other groups, at the end of our experiment we found that the ratio of insulin positive cell in normal group which have been measured by immunohisto chemistry was conspicuous higher than other each group, the ratio of mobilization plus induction group is lower than normal group, but higher than DM group and mobilization group, and there has no significant difference between mobilization and DM group. We found insulin positive cell expression in liver but no expression in spleen in other groups. We did not find any expression of insulin positive cell in liver and spleenConclusionCombination therapy with Epidermal Growth Factor and Gastrin could induce differention of bone marrow stem cell into insulin-producing cell, and insulin-producing cells have curiat effect on rat diabetes.