Oreocnide integrifolia Flavonoids Augment Reprogramming for Islet Neogenesis and β-Cell Regeneration in Pancreatectomized BALB/c Mice

Agents which can either trigger proliferation of β-cells or induce neogenesis of β-cells from precursors would be of pivotal role in reversing diabetic manifestations. We examined the role of flavonoid rich fraction (FRF) of Oreocnide integrifolia leaves using a mice model of experimental regeneration. BALB/c mice were subjected to ~70% pancreatectomy (Px) and supplemented with FRF for 7, 14, and 21 days after pancreatectomy. Px animals displayed increased blood glucose levels and decreased insulin titres which were ameliorated by FRF supplementation. FRF-treated mice demonstrated prominent newly formed islets budding off from ducts and depicting increased BrdU incorporation. Additionally, transcripts levels of Ins1/2, Reg-3α/γ, Ngn-3, and Pdx-1 were upregulated during the initial 1 week. The present study provides evidence of a nutraceutical contributing to islet neogenesis from ductal cells as the mode of β-cell regeneration and a potential therapeutic for clinical trials in management of diabetic manifestations

Antioxidant rich flavonoids from Oreocnide integrifolia enhance glucose uptake and insulin secretion and protects pancreatic β-cells from streptozotocin insult

<p>Abstract</p> <p>Background</p> <p>Insulin deficiency is the prime basis of all diabetic manifestations and agents that can bring about insulin secretion would be of pivotal significance for cure of diabetes. To test this hypothesis, we carried out bioactivity guided fractionation of <it>Oreocnide integrifolia </it>(Urticaceae); a folklore plant consumed for ameliorating diabetic symptoms using experimental models.</p> <p>Methods</p> <p>We carried out bioassay guided fractionation using RINmF and C2C12 cell line for glucose stimulated insulin secretion (GSIS) and glucose uptake potential of fractions. Further, the bioactive fraction was challenged for its GSIS in cultured mouse islets with basal (4.5 mM) and stimulated (16.7 mM) levels of glucose concentrations. The Flavonoid rich fraction (FRF) was exposed to 2 mM streptozotocin stress and the anti-ROS/RNS potential was evaluated. Additionally, the bioactive fraction was assessed for its antidiabetic and anti-apoptotic property <it>in-vivo </it>using multidose streptozotocin induced diabetes in BALB/c mice.</p> <p>Results</p> <p>The results suggested FRF to be the most active fraction as assessed by GSIS in RINm5F cells and its ability for glucose uptake in C2C12 cells. FRF displayed significant potential in terms of increasing intracellular calcium and cAMP levels even in presence of a phosphodiesterase inhibitor, IBMX in cultured pancreatic islets. FRF depicted a dose-dependent reversal of all the cytotoxic manifestations except peroxynitrite and NO formation when subjected <it>in-vitro </it>along with STZ. Further scrutinization of FRF for its <it>in-vivo </it>antidiabetic property demonstrated improved glycemic indices and decreased pancreatic β-cell apoptosis.</p> <p>Conclusions</p> <p>Overall, the flavonoid mixture has shown to have significant insulin secretogogue, insulinomimetic and cytoprotective effects and can be evaluated for clinical trials as a therapeutant in the management of diabetic manifestations.</p

Induction of regulatory T cells: A role for probiotics and prebiotics to suppress autoimmunity.

Regulatory T cells (Tregs) are comprised of a heterogeneous population of cells that play a vital role in suppressing inflammation and maintaining immune tolerance. Given the crucial role of Tregs in maintaining immune homeostasis, it is probably not surprising that many microbial species and their metabolites have the potential to induce Tregs. There is now great interest in the therapeutic potential of probiotics and prebiotics based strategies for a range of autoimmune disorders. This review will summarise recent findings concerning the role of probiotics and prebiotics in induction of Tregs to ameliorate the autoimmune conditions. In addition, the article is focused to explain the different mechanisms of Treg induction and function by these probiotics and prebiotics, based on the available studies till date. The article further proposes that induction of Tregs by probiotics and prebiotics could lead to the development of new therapeutic approach towards curbing the autoimmune response and as an alternative to detrimental immunosuppressive drugs

Increased Tumor Necrosis Factor (TNF)-α and Its Promoter Polymorphisms Correlate with Disease Progression and Higher Susceptibility towards Vitiligo

<div><h3>Abstract</h3><p>Tumor Necrosis Factor (TNF)-α, is a paracrine inhibitor of melanocytes, which plays a critical role in the pathogenesis of several autoimmune diseases including vitiligo, as abnormal immune responses have frequently been observed in vitiligo patients. Moreover, vitiligo patients show higher lesion levels of TNF-α. Genetic polymorphisms in the promoter region of <em>TNF-α</em> are involved in the regulation of its expression. The present study explores <em>TNF</em>-α promoter polymorphisms and correlates them with <em>TNF-α</em> transcript and protein levels in vitiligo patients and controls of Gujarat along with its effect on disease onset and progression. PCR-RFLP technique was used for genotyping of these polymorphisms in 977 vitiligo patients and 990 controls. TNF-α transcript and protein levels were measured by Real time PCR and ELISA respectively. The genotype and allele frequencies for the investigated polymorphisms were significantly associated with vitiligo patients. The study revealed significant increase in <em>TNF</em>-α transcript and protein levels in vitiligo patients compared to controls. In particular, haplotypes: AATCC, AACCT, AGTCT, GATCT, GATCC and AGCCT were found to increase the TNF-α levels in vitiligo patients. Analysis of TNF-α levels based on the gender and disease progression suggests that female patients and patients with active vitiligo had higher levels of TNF-α. Also, the TNF-α levels were high in patients with generalized vitiligo as compared to localized vitiligo. Age of onset analysis of the disease suggests that the haplotypes: AACAT, AACCT, AATCC and AATCT had a profound effect in the early onset of the disease. Moreover, the analysis suggests that female patients had an early onset of vitiligo. Overall, our results suggest that <em>TNF</em>-α promoter polymorphisms may be genetic risk factors for susceptibility and progression of the disease. The up-regulation of <em>TNF</em>-α transcript and protein levels in individuals with susceptible haplotypes advocates the crucial role of TNF-α in autoimmune pathogenesis of vitiligo.</p> </div

Relative gene expression of <i>TNF</i>-α with respect to promoter genotypes in controls and vitiligo patients.

<p>(<b>A</b>) Expression of <i>TNF</i>-α transcripts with respect to −238 G/A genotypes in 157 vitiligo patients and 174 controls, as suggested by Mean ΔCp. (<b>B</b>) Expression of <i>TNF</i>-α transcripts with respect to −308 G/A genotypes in 157 vitiligo patients and 174 controls, as suggested by Mean ΔCp. (<b>C</b>) Expression of <i>TNF</i>-α transcripts with respect to −857 C/T genotypes in 157 vitiligo patients and 174 controls, as suggested by Mean ΔCp. (<b>D</b>) Expression of <i>TNF</i>-α transcripts with respect to −863 C/A genotypes in 157 vitiligo patients and 174 controls, as suggested by Mean ΔCp. (<b>E</b>) Expression of <i>TNF</i>-α transcripts with respect to −1031 T/C genotypes in 157 vitiligo patients and 174 controls, as suggested by Mean ΔCp.</p

Distribution of haplotypes frequencies for <i>TNF</i>-α promoter polymorphisms among localized vitiligo patients and controls.

<p>CI represents Confidence Interval,</p><p>(Frequency <0.03 in both control & case has been dropped and was ignored in analysis).</p

Serum TNF-α levels in controls and vitiligo patients.

<p>(<b>A</b>) Comparison of sTNF-α levels (pg/ml) in 236 controls, 214 vitiligo patients, 158 generalized vitiligo patients and 56 localized vitiligo patients, as determined by ELISA. (<b>B</b>)<b>.</b> Comparison of sTNF-α levels (pg/ml) with respect to <i>TNF</i>-α promoter haplotypes in 214 vitiligo patients and 236 controls, as determined by ELISA. (<b>C</b>) Comparison of sTNF-α levels (pg/ml) with respect to activity of the disease in 150 patients with active vitiligo and 64 patients with stable vitiligo, as determined by ELISA. (<b>D</b>) Comparison of sTNF-α levels (pg/ml) with respect to gender differences in 97 male patients and 117 female patients with vitiligo, as determined by ELISA.</p