The Mechanism Of Jiangtangsanhuang Tablet On The Apoptosis Of Rat Beta Cells Induced By Interleukin-1β

ObjectiveDiabetes Mellitus, the issue of common concern worldwide, has been characterized by pandemic risk, financial burden, tendency of lower age and severely affected health. It is a systemic metabolic disease often associated with symptoms such as weight loss, polyphagia, polydipsia and polyuria. Without appropriate treatment and therapeutic intervention, diabetic patients may develop multiple complications, including cardiovascular disease, renal failure and neuropathy and compromised wound healing.Interleukin-1β as one of Cytokines have been implicated as effector molecules that participate in the destruction of β cells leading to the development of type1and type2diabetic mellitus. Interleukin-1β produces excess nitric oxide (NO) by the inducible form of nitric oxide synthase (iNOS) and causes apoptosis by the increase of capase-3for rat pancreatic β cells. It is reported that MAPK pathways have been also reported to be involved in the regulation of cytokine-induced-cell death, interleukin-1β could doubt to activate the p38Kinase and the c-Jun N-terminal kinase in pancreatic β cells to control the express of nitric oxide synthase.We have investigated the preventive mechanism of Jiangtang sanhuang Tablet protects β cells from IL-1β to broaden the field of clinical application.MethodsWe made use of bile reflux perfusion, removal perfused pancreata, cell culture, MTT assay, Nitrite measurement, RNA isolation and real-time PCR of iNOS, caspase-3to complete our experiment.ResultsIt was obtained effective and purified islets by Bile reflux perfusion and Ficoll-400. To examine the function of free islets, the insulin secretion assay was performed in the presence of either2.8or16.7mM D-glucose. Static incubation of islets at16.7mM glucose for2h resulted in a2.01-fold increase in insulin secretion by islets. Therefore, free islets by this way did not obviously affect the glucose-stimulated insulin secretion.A24h incubation of the rat islets with IL-1β resulted increase in NO production, meanwhile, Real-time PCR data showed that iNOSmRNA and caspase-3mRNA were markedly increased by IL-1β, and Western blot data showed increased P38and JNK could promote the production of iNOS which resulted increase in NO production. Islets pretreated with Jiangtangsanhuang Tablet partly abolished the effects of IL-1β and resulted in control levels of NO production, iNOS and caspase-3expression, but relied on its concentration.ConclusionIn the present study, we have evidences that the preventive mechanism of Jiangtangsanhuang Tablet protects β cells from IL-1β induced β-cell damage by suppressing iNOS and caspase-3expression in rats’islets.