A Small Molecule Swertisin fromEnicostemma littoraleDifferentiates NIH3T3 Cells into Islet-Like Clusters and Restores Normoglycemia upon Transplantation in Diabetic Balb/c Mice

Abstract

Aim. Stem cell therapy is one of the upcoming therapies for the treatment of diabetes. Discovery of potent differentiating agents is a prerequisite for increasing islet mass. The present study is an attempt to screen the potential of novel small biomolecules for their differentiating property into pancreatic islet cells using NIH3T3, as representative of extra pancreatic stem cells/progenitors. Methods. To identify new agents that stimulate islet differentiation, we screened various compounds isolated from Enicostemma littorale using NIH3T3 cells and morphological changes were observed. Characterization was performed by semiquantitative RT-PCR, Q-PCR, immunocytochemistry, immunoblotting, and insulin secretion assay for functional response in newly generated islet-like cell clusters (ILCC). Reversal of hyperglycemia was monitored after transplanting ILCC in STZ-induced diabetic mice. Results. Among various compounds tested, swertisin, an isolated flavonoid, was the most effective in differentiating NIH3T3 into endocrine cells. Swertisin efficiently changed the morphology of NIH3T3 cells from fibroblastic to round aggregate cell cluster in huge numbers. Dithizone (DTZ) stain primarily confirmed differentiation and gene expression studies signified rapid onset of differentiation signaling cascade in swertisin-induced ILCC. Molecular imaging and immunoblotting further confirmed presence of islet specific proteins. Moreover, glucose induced insulin release (in vitro) and decreased fasting blood glucose (FBG) (in vivo) in transplanted diabetic BALB/c mice depicted functional maturity of ILCC. Insulin and glucagon expression in excised islet grafts illustrated survival and functional integrity. Conclusions. Rapid induction for islet differentiation by swertisin, a novel herbal biomolecule, provides low cost and readily available differentiating agent that can be translated as a therapeutic tool for effective treatment in diabetes.

Anti-apoptotic and cytoprotective effect ofEnicostemma littoraleagainst oxidative stress in Islets of Langerhans

“…Enicostemma littorale, a perennial herb, has been used for therapy of diabetes mellitus in India as a folk medicine (Saranya et al 2013). Very recently we also demonstrated that EL MeOH extract possess potential islet neogenic activity from tissue specific stem/progenitor cells, which was later confirmed by identifying the active ingredient swertisin for this islet neogenic potential in a another recent report from us (Dadheech et al 2010(Dadheech et al , 2013. There are also other reports where decrease in insulin resistance, normalization of fat and carbohydrate metabolism by EL were observed in both experimental cellular and rodent models (Maroo et al 2002;Vasu et al 2003;Patel et al 2013).…”

Swertiamarin: An Active Lead fromEnicostemma littoraleRegulates Hepatic and Adipose Tissue Gene Expression by Targeting PPAR-γand Improves Insulin Sensitivity in Experimental NIDDM Rat Model

“…Further, the author's research group is continuously involved in exploring the wide spectrum hidden potentials of this plant for islet neogenesis and various diabetic complications [15, 22, 23]. Qualitative analyses of EL have demonstrated the presence of flavonoids and secoiridoid glycosides.…”

“…Our lab has well documented Enicostemma littorale Blume owing antioxidant, hypolipidemic, and antidiabetic activities both in animals and human diabetic patients [13, 14]. Apart from antidiabetic activity, islet neogenic property of swertisin and normoglycemia in a diabetic rat are also reported [15]. Hence, it can be presumed that the EL extract potentially owes varied beneficiary activities due to the presence of many compounds within it.…”

Differentiation of stem cells into insulin‐producing cells under the influence of nanostructural polyoxometalates

“…The most studied transcription factors (pancreatic and duodenal homeobox factor-1, neurogenin 3 and musculoaponeurotic fibrosarcoma oncogene family protein A) are expressed in β-cell precursors, which have a role in inducing differentiation and in maintaining the functionality of the β-cells (Kaneto et al, 2007). The most important obstacles to the clinical use of these transcription factors, as well as of the above-mentioned hormones and growth factors, are represented by their high costs and low efficiency in obtaining functional βcells clusters (Dadheech et al, 2013). Thus, in these circumstances, POMs have been proposed as alternative, effective molecules.…”