| Diabetes Mellitus is one of the most common endocrine and metabolic disease, and serious hazard to human health. In recent years, the morbidity and mortality of diabetes mellitus showed a rising trend. The disadvantage of chemical treatment, including toxicol medicine and can not be effectively prevent the complications of diabetes mellitus. Therefore, presuming hypoglycemic active ingredients from natural plant has become an important way to treat the diabetes mellitus. In recent years, the hypoglycemic effect of plant polysaccharides have been found, and is expected to develop the new phytochemical antidiabetic medicine form natural plant. To date, the domestic, foreign papers, patents reported of the hypoglycemic effect on Sphallerocarpus gracilis polysaccharide (SGP) have not been found. Thus, the aim of this study is to research the prevention of SGP, treatment of diabetes mellitus, and the quantity and function deteriorates of β cell to explore the mechanism of prevention and treatment of diabetes mellitus. And provide a theoretical basis for the following research of SGP hypoglycemic natural product.SGP is extracted from the Sphallerocarpus gracilis,which is the characteristics of Gansu plants. Alloxan induce experimental diabetes, graded concentrations of SGP were effected into the alloxan iniured RINm5F cells. SOD and NOS activity, NO and MDA production were measured by colorimetrically.The cells viability was determined by MTT assay. The amount of glucose-stimulated insulin secretion was assayed by radioimmunoassay(RIA). Real-time qPCR is applied to detect the caspase-3, bax, bcl-2, pdx-1, insulin mRNA expression. Western-blot is applied to detect the expression of Caspase-3and PDX-1.Results show that:1The SGP could repair free radical damage on RINm5F cellsThe SGP have a protective effect on free radical damage by alloxan in RINm5F cells. It is possibly through that SGP could inhibite NOS activity in RINm5F cells, reduce the production of NO, improve SOD activity, decrease the content of MDA, enhance ability to scavenge free radicals, improve antioxidant capacity.2The SGP inhibite the apoptosis on alloxan injured RINm5F cellsThe SGP could enhance the cells viability obviously, and with the increase of SGP concentration, the cells viability increase gradually, with the increase of the duration of action, the cells viability decline gradually. It is possibly through that SGP increase significantly bcl-2mRNA expression, and decrease significantly bax mRNA expression, than finally decrease caspase-3gene transcription and protein expression, inhibite the apoptosis by alloxan in RINm5F cells.3The SGP repair dysfunction of alloxan injured RINm5F cellsIn the presence of2.8mmol/L and16.7mmol/L glucose, SGP increased glucose-stimulated insulin secretion in RINm5F cells in a dose-dependent manner, and high-glucose-stimulated insulin secretion is more than lower. It is possibly through that SGP could increase significantly pdx-1gene transcription and protein expression, and increase insulin secretion, to repair dysfunction of alloxan injured RINm5F cells.Conclusion: the SGP could protect and repaire alloxan injuryed RINm5F cells by(1) improved antioxidant capacity, reduced lipid oxidation productions (2) activated bcl-2mRNA transciption, and inhibited bax mRNA transciption, eventually inhibited caspase-3gene transcription and Caspase-3protein expression, inhibited bata cell apoptosis (3) activated the transcription of pdx-1and insulin mRNA, activated PDX-1protein expression, improved insulin secretion level and repaired the injured RINm5F cells. |
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