Stimulation of Apolipoprotein A-IV expression in Caco-2/TC7 enterocytes and reduction of triglyceride formation in 3T3-L1 adipocytes by potential anti-obesity Chinese herbal medicines

<p>Abstract</p> <p>Background</p> <p>Chinese medicine has been proposed as a novel strategy for the prevention of metabolic disorders such as obesity. The present study tested 17 Chinese medicinal herbs were tested for their potential anti-obesity effects.</p> <p>Methods</p> <p>The herbs were evaluated in terms of their abilities to stimulate the transcription of Apolipoprotein A-IV (ApoA-IV) in cultured Caco-2/TC7 enterocytes. The herbs that showed stimulating effects on ApoA-IV transcription were further evaluated in terms of their abilities to reduce the formation of triglyceride in differentiated 3T3-L1 adipocytes.</p> <p>Results</p> <p>ApoA-IV transcription was stimulated by <it>Rhizoma Alismatis </it>and <it>Radix Angelica Sinensis </it>in a dose- and time-dependent manner in cultured Caco-2/TC7 cells. Moreover, these two herbs reduced the amount of triglyceride in differentiated 3T3-L1 adipocytes.</p> <p>Conclusion</p> <p>The results suggest that <it>Rhizoma Alistmatis </it>and <it>Radix Angelica Sinensis </it>may have potential anti-obesity effects as they stimulate ApoA-IV transcription and reduce triglyceride formation.</p

Kaempferol as a flavonoid induces osteoblastic differentiation via estrogen receptor signaling

<p>Abstract</p> <p>Background</p> <p>Flavonoids, a group of compounds mainly derived from vegetables and herbal medicines, chemically resemble estrogen and some have been used as estrogen substitutes. Kaempferol, a flavonol derived from the rhizome of <it>Kaempferia galanga </it>L., is a well-known phytoestrogen possessing osteogenic effects that is also found in a large number of plant foods.</p> <p>The herb <it>K. galanga </it>is a popular traditional aromatic medicinal plant that is widely used as food spice and in medicinal industries. In the present study, both the estrogenic and osteogenic properties of kaempferol are evaluated.</p> <p>Methods</p> <p>Kaempferol was first evaluated for its estrogenic properties, including its effects on estrogen receptors. The osteogenic properties of kaempferol were further determined its induction effects on specific osteogenic enzymes and genes as well as the mineralization process in cultured rat osteoblasts.</p> <p>Results</p> <p>Kaempferol activated the transcriptional activity of pERE-Luc (3.98 ± 0.31 folds at 50 μM) and induced estrogen receptor α (ERα) phosphorylation in cultured rat osteoblasts, and this ER activation was correlated with induction and associated with osteoblast differentiation biomarkers, including alkaline phosphatase activity and transcription of osteoblastic genes, <it>e.g</it>., type I collagen, osteonectin, osteocalcin, Runx2 and osterix. Kaempferol also promoted the mineralization process of osteoblasts (4.02 ± 0.41 folds at 50 μM). ER mediation of the kaempferol-induced effects was confirmed by pretreatment of the osteoblasts with an ER antagonist, ICI 182,780, which fully blocked the induction effect.</p> <p>Conclusion</p> <p>Our results showed that kaempferol stimulates osteogenic differentiation of cultured osteoblasts by acting through the estrogen receptor signaling.</p

Controlled induction of human pancreatic progenitors produces functional beta‐like cells in vitro

Directed differentiation of human pluripotent stem cells into functional insulin‐producing beta‐like cells holds great promise for cell replacement therapy for patients suffering from diabetes. This approach also offers the unique opportunity to study otherwise inaccessible aspects of human beta cell development and function in vitro. Here, we show that current pancreatic progenitor differentiation protocols promote precocious endocrine commitment, ultimately resulting in the generation of non‐functional polyhormonal cells. Omission of commonly used BMP inhibitors during pancreatic specification prevents precocious endocrine formation while treatment with retinoic acid followed by combined EGF/KGF efficiently generates both PDX1+ and subsequent PDX1+/NKX6.1+ pancreatic progenitor populations, respectively. Precise temporal activation of endocrine differentiation in PDX1+/NKX6.1+ progenitors produces glucose‐responsive beta‐like cells in vitro that exhibit key features of bona fide human beta cells, remain functional after short‐term transplantation, and reduce blood glucose levels in diabetic mice. Thus, our simplified and scalable system accurately recapitulates key steps of human pancreas development and provides a fast and reproducible supply of functional human beta‐like cells.SynopsisFocusing on developmental mechanisms, the results of this study further accelerate successful differentiation of human ESCs into functional pancreatic beta cells.Exclusion of commonly used BMP inhibitors during human embryonic stem cell to pancreatic progenitor differentiation prevents precocious endocrine induction.Sequential exposure of foregut cells to retinoic acid followed by combined EGF/KGF treatment establishes highly pure PDX1+ and PDX1+/NKX6.1+ progenitor populations, respectively.Precise temporal induction of endocrine differentiation in PDX1+/NKX6.1+ progenitors, but not in PDX1+/NKX6.1− progenitors, results in the generation of functional beta‐like cells in vitro.Beta‐like cells exhibit key features of bona fide human beta cells, remain functional after short‐term transplantation, and reduce blood glucose levels in diabetic mice.Focusing on developmental mechanisms, the results of this study further accelerate successful differentiation of human ESCs into functional pancreatic beta cells.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/111932/1/embj201591058.reviewer_comments.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/111932/2/embj201591058.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/111932/3/embj201591058-sup-0001-FigsS1-S4.pd

Functional Analysis of Genes Involved in Regulating Stem Cell Maintenance and Differentiation in the Freshwater Planarian Schmidtea Mediterranea

131 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2007.Freshwater planarians have been a classic model for regeneration studies for more than 250 years. The great regenerative abilities of planarians rely on neoblasts, adult stem cells maintained throughout the animal's life. This thesis reports an analysis of genes that regulate stem cell maintenance and differentiation in planarian Schmidtea mediterranea. A member of the Bruno-like family of RNA binding proteins was found to be critical for neoblast self-renewal in the planarian Schmidtea mediterranea. Smed-bruno-like (bruli) mRNA and protein are expressed in the neoblasts and in the central nervous system. Immunofluorecense with antibodies recognizing SMEDWI-1 and Historie H4 (monomethyl-K20) and other cell cycle markers showed that the neoblast population was eliminated from both regenerating and intact planarians following smed-bruli RNA interference, leading to the death of the animals. These results suggest that Bruli is required for stem cell maintenance. We further characterized expression of SMEDWI-1 in somatic stem cells and germ cells. SMEDWI-1 protein is expressed in neoblasts and their differentiating progeny in both asexual and sexual planarians. In sexual planarians, SMEDWI-1 expression is detected in spermatogonia and spermatocytes as well as oogonia. SMEDWI-1 antibodies also recognize the presumptive germ cells in asexual planarians that reproduce exclusively by fission. However, SMEDWI-1 protein is dispensable for the development, maintenance and regeneration of testes in sexual planarians, probably due to the redundancy of other PIWI family members expressed in germ cells. Finally, we examined the differentiation of neoblasts in intact and Smed-slit RNAi planarians. BraU incorporation and immunofluorescent labeling with cell type-specific markers were used to label proliferating neoblasts and follow the fates of their progeny into various differentiated cell types, including marginal gland cells, photoreceptors, and neurons. In slit RNAi animals, neoblasts differentiate into ectopic neural tissues that form ganglia-like structures along the midline. Thus, Smed-slit may function to regulate neoblast differentiation in addition to acting as a repulsive cue in axon guidance.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD

Functional Analysis of Genes Involved in Regulating Stem Cell Maintenance and Differentiation in the Freshwater Planarian Schmidtea Mediterranea

131 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2007.Freshwater planarians have been a classic model for regeneration studies for more than 250 years. The great regenerative abilities of planarians rely on neoblasts, adult stem cells maintained throughout the animal's life. This thesis reports an analysis of genes that regulate stem cell maintenance and differentiation in planarian Schmidtea mediterranea. A member of the Bruno-like family of RNA binding proteins was found to be critical for neoblast self-renewal in the planarian Schmidtea mediterranea. Smed-bruno-like (bruli) mRNA and protein are expressed in the neoblasts and in the central nervous system. Immunofluorecense with antibodies recognizing SMEDWI-1 and Historie H4 (monomethyl-K20) and other cell cycle markers showed that the neoblast population was eliminated from both regenerating and intact planarians following smed-bruli RNA interference, leading to the death of the animals. These results suggest that Bruli is required for stem cell maintenance. We further characterized expression of SMEDWI-1 in somatic stem cells and germ cells. SMEDWI-1 protein is expressed in neoblasts and their differentiating progeny in both asexual and sexual planarians. In sexual planarians, SMEDWI-1 expression is detected in spermatogonia and spermatocytes as well as oogonia. SMEDWI-1 antibodies also recognize the presumptive germ cells in asexual planarians that reproduce exclusively by fission. However, SMEDWI-1 protein is dispensable for the development, maintenance and regeneration of testes in sexual planarians, probably due to the redundancy of other PIWI family members expressed in germ cells. Finally, we examined the differentiation of neoblasts in intact and Smed-slit RNAi planarians. BraU incorporation and immunofluorescent labeling with cell type-specific markers were used to label proliferating neoblasts and follow the fates of their progeny into various differentiated cell types, including marginal gland cells, photoreceptors, and neurons. In slit RNAi animals, neoblasts differentiate into ectopic neural tissues that form ganglia-like structures along the midline. Thus, Smed-slit may function to regulate neoblast differentiation in addition to acting as a repulsive cue in axon guidance.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD

DNA methylation directs functional maturation of pancreatic β cells

Pancreatic β cells secrete insulin in response to postprandial increases in glucose levels to prevent hyperglycemia and inhibit insulin secretion under fasting conditions to protect against hypoglycemia. β cells lack this functional capability at birth and acquire glucose-stimulated insulin secretion (GSIS) during neonatal life. Here, we have shown that during postnatal life, the de novo DNA methyltransferase DNMT3A initiates a metabolic program by repressing key genes, thereby enabling the coupling of insulin secretion to glucose levels. In a murine model, β cell-specific deletion of Dnmt3a prevented the metabolic switch, resulting in loss of GSIS. DNMT3A bound to the promoters of the genes encoding hexokinase 1 (HK1) and lactate dehydrogenase A (LDHA) - both of which regulate the metabolic switch - and knockdown of these two key DNMT3A targets restored the GSIS response in islets from animals with β cell-specific Dnmt3a deletion. Furthermore, DNA methylation-mediated repression of glucose-secretion decoupling genes to modulate GSIS was conserved in human β cells. Together, our results reveal a role for DNA methylation to direct the acquisition of pancreatic β cell function

Characterization, Molecular Docking, and In Vitro Dissolution Studies of Solid Dispersions of 20(S)-Protopanaxadiol

In this study, we prepared solid dispersions (SDs) of 20(S)-protopanaxadiol (PPD) using a melting-solvent method with different polymers, in order to improve the solubility and dissolution performance of drugs with poor water solubility. The SDs were characterized via differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), and molecular docking and dynamics study. DSC and PXRD results indicated that PPD crystallinity in SDs was significantly reduced, and that the majority of PPD is amorphous. No interaction was observed between PPD and polymers on FTIR and NMR spectra. Molecular docking and dynamic calculations indicated that the PPD molecule localized to the interpolated charged surface, rather than within the amorphous polymer chain network, which might help prevent PPD crystallization, consequently enhancing the PPD dispersion in polymers. An in vitro dissolution study revealed that the SDs considerably improved the PPD dissolution performance in distilled water containing 0.35% Tween-80 (T-80). Furthermore, among three PPD-SDs formulations, Poloxamer188 (F68) was the most effective in improving the PPD solubility and was even superior to the mixed polymers. Therefore, the SD prepared with F68 as a hydrophilic polymer carrier might be a promising strategy for improving solubility and in vitro dissolution performance. F68-based SD, containing PPD with a melting-solvent preparation method, can be used as a promising, nontoxic, quick-release, and effective intermediate for other pharmaceutical formulations, in order to achieve a more effective drug delivery

Edible Bird's Nest, an Asian Health Food Supplement, Possesses Moisturizing Effect by Regulating Expression of Filaggrin in Skin Keratinocyte

Edible bird's nest (EBN) has been consumed as a Chinese delicacy for hundreds of years; the functions of which have been proposed to prevent lung disease, strengthen immune response, and restore skin youthfulness. To support the skin function of EBN, the water extract and the enzymatic digest of EBN with enriched digested peptides were tested in cultured keratinocyte, HaCaT cell line. The effects of EBN extract and digest in inducing proteins crucial for skin moisturizing were determined in both in vitro and ex vivo models. In cultured keratinocytes, the expressions of S100-fused type proteins contributing to skin barrier function in the stratum corneum, e.g. filaggrin and filaggrin-2, were determined in both mRNA and protein levels, which were markedly induced in the treatment of EBN extract or digest. The EBN-induced gene transcriptions of filaggrin and filaggrin-2 were mediated by activation of p38 MAPK pathway and various transcription factors, e.g. GATA3, PPAR alpha, PPAR beta, and PPAR gamma: these transcriptional factors were markedly activated by the digested products of EBN, as compared to the extract, in cultured keratinocytes. By using atomic force microscopy (AFM), the EBN-treated keratinocyte was shown to have more liquid-like morphology, as compared to a control cell. The EBN digest showed better induction on these moisturizing effects as compared to the extract. These lines of evidence therefore suggested the water moisturizing effect of EBN in skin function