| [AIM] Type I Diabetes is an autoimmune disease. Although Insulin can improve patient's clinical syndrome but can't prevent development of disease and appearance of complex. The patients need to accept insulin injected for all life time. In recent years, achievement on stem cell research provides a possibility for us to induce stem cells to differentiate into islet cell and promote neogenesis of pancreatic islet. It presents us a promising hope to cure type 1 diabetes. In this study, we investigate the feasibility of granulocyte colony-stimulating factor (G-CSF) on therapy of the mice with diabetes by mobilizing autologous bone marrow stem cells and then directly migrating to pancreas in vivo under biological condition.[METHODS] The BALB/C female mice with 22-24g were randomly divided into six groups.(l) normal group that is normal mice injected with PBS subcutaneously; (2) diabetic model group that is normal mice injected with streptozocin 250 to300 mg/kg /day; intraperitonealy (3) insulin-treated group that is diabetic mice injected with insulin 2IU/kg/day subcutaneously. (4) G-CSF-treated group that is diabetic mice injected with G-CSF 20ug/kg/day subcutaneously; (5) G-CSF plus insulin-treated group that is diabetic mice injected with insulin2JU /kg/day subcutaneously in addition to G-CSF 20ug /kg/day; (6) G-CSF toxicity test group that is normal mice injected with G-CSF20ug/kg/day.subcutaneously There were nine mice in each group, all of which were treated continuously for sixty days. Blood glucose levels weredetected at day 15, 30, 60 post-therapy. Both urinary glucose levels and body weight were measured daily. The mice were killed for both the pathology and immunohistochemistry at day 30, 60 post-therapy. [RESULTS]1. Establishment of diabetic modelTo set up diabetic model, normal mice were injected with streptozocin. After 6 to 9 days, blood glucose and urinary glucose of mice increased, at the same time their weight decreased continuously. Their general situations were poor, their movements were inactive and hair had no shine. The amount of urine was obviously more than that of the normal mice. All mice died within 40 days. The results from pathology showed that the structure of pancreatic islet was destroyed severely and spleen of diabetic mice was as half as that of normal mice.2. The curative effects of insulin treatment in diabetic model mice Insulin is basic drug for treatment of patients with diabetes. Beforestudied the effect of G-CSF for diabetes treatment, we observed the effect of insulin in diabetic model mice as a control. After insulin therapy, the urine amount, glucose levels in urine and blood of insulin-treated mice decreased and their weight increased. The 60 percent of mice was living at day 60 post therapy. But insulin-treated mice were still inactive, their spleen was smaller and the lesion of islet cells had no improvement compared with diabetic mice. In addition, there was no CD34+ cell detected by immunohistochemistry in pancreatic islet. This data suggested that insulin has on role on neogenesis of pancreatic islet although it could decrease glucose level in urine and blood of mice.3. The curative effects of G-CSF treatment in diabetic model mice The general situations of diabetic mice had improved following G-CSFtherapy. Their weight elevated and their hair shined. They became active in their movements, there were obvious decreased blood glucose before dinner, All mice vrere living at day 60 post therapy. The size of their spleenwas almost normol,The result from pathology showed that some part of destroyed pancreatic islet had recovered. But there were no obvious decreased blood glucose after dinner and urinary glucose and the amount of urine were almost as much as that of the mice in diabetic model mice. These data demonstrated that G-CSF could promote the neogenesis of pancreatic islet and had more effect for enhancing living quality and long term living rate, although it's effect for reducing urine and blood glucose level wasn't obvious, It suggeste that G-CSF will be a novel drug for diabetes treatment.4. The united curative effects of insulin and G-CSF treatment on diabetic model miceThe results above indicated that neither insulin nor G-CSF alone had some disadvantage for diabetes therapy. In order to explore new method for diabetes treatment, we had observed the united curative effects of insulin and G-CSF for diabetes treatment. The result demonstrated that insulin and G-CSF possessed compensative effect each other. They did not only decrease glucose level in blood and urine significantly but also improve the general situation of mice (such as, shined hair, increased weight and activated movements) and elevate the living rate (100% at day 60 post therapy). More exiting, result from pathology and immunohistochemistry indicated that pancreatic islet almost completely recovered from lesion and that CD34+ stem cells were found within islet and some of them had differentiated into insulin-produced islet cells. This suggested that united application of insulin and G-CSF will be an effective method for diabetes treatment.5. G-CSF toxicity testThe mice injected with 20ug/kg/day of G-CSF for 60 had no any side effect. ICONCLUSION AND SINGIFICANCEJ1. It was found that G-CSF could promote the neogenesis of pancreaticislet and had more effect for enhancing living quality and long term living rate of mice. It suggested that G-CSF will be a novel drug for diabetes treatment.2. It was demonstrated that united use of insulin and G-CSF possessed compensative effect each other. They did not only decrease glucose level in blood and urine of diabetic mice significantly, but also improve the general situation of mice and elevate the living rate and promote the neogenesis of pancreatic islet. This suggested that united application of insulin and G-CSF will be an effective method for diabetes treatment. |
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