Bone resorption by osteoclasts is a critical step in bone remodeling,a process important for maintaining bone homeostasis and repairing injured bone.We previously identified a bone marrow mesenchymal subpopulation,mar...Bone resorption by osteoclasts is a critical step in bone remodeling,a process important for maintaining bone homeostasis and repairing injured bone.We previously identified a bone marrow mesenchymal subpopulation,marrow adipogenic lineage precursors(MALPs),and showed that its production of RANKL stimulates bone resorption in young mice using Adipoq-Cre.To exclude developmental defects and to investigate the role of MALPs-derived RANKL in adult bone,we generated inducible reporter mice(Adipoq-CreER Tomato)and RANKL deficient mice(Adipoq-CreER RANKLflox/flox,iCKO).Single cell-RNA sequencing data analysis and lineage tracing revealed that Adipoq+cells contain not only MALPs but also some mesenchymal progenitors capable of osteogenic differentiation.In situ hybridization showed that RANKL mRNA is only detected in MALPs,but not in osteogenic cells.RANKL deficiency in MALPs induced at 3 months of age rapidly increased trabecular bone mass in long bones as well as vertebrae due to diminished bone resorption but had no effect on the cortical bone.Ovariectomy(OVX)induced trabecular bone loss at both sites.RANKL depletion either before OVX or at 6 weeks post OVX protected and restored trabecular bone mass.Furthermore,bone healing after drill-hole injury was delayed in iCKO mice.Together,our findings demonstrate that MALPs play a dominant role in controlling trabecular bone resorption and that RANKL from MALPs is essential for trabecular bone turnover in adult bone homeostasis,postmenopausal bone loss,and injury repair.展开更多
OBJECTIVE:To investigate the effect of Shisiwei Jianzhong decoction(十四味建中汤,SJD)on non-severe aplastic anemia(NSAA).METHODS:Bone marrow mesenchymal stem cells(BMSCs)were isolated from bone marrow samples of 15 NS...OBJECTIVE:To investigate the effect of Shisiwei Jianzhong decoction(十四味建中汤,SJD)on non-severe aplastic anemia(NSAA).METHODS:Bone marrow mesenchymal stem cells(BMSCs)were isolated from bone marrow samples of 15 NSAA patients and 3 healthy controls.Cells were treated with gradient concentrations of SJD,and a portion was transfected with a vector overexpressing the nuclear factor of activated T cells,cytoplasmic 4(NFATC4).Cell viability and apoptosis were detected by cell counting kit-8 and flow cytometry,respectively.After adipogenic differentiation induction,lipid droplet formation in BMSCs was examined by Oil Red O staining.The expression of NFATC4,peroxisome proliferator-activated receptor gamma(PPARG),fatty acid-binding protein 4(FABP4),peroxisome proliferator-activated receptor-gamma coactivator(PGC-1α),and acetylated PGC-1αwas measured by quantitative real-time polymerase chain reaction or Western blot.RESULTS:SJD significantly increased the viability and decreased the apoptosis of NSAA-derived BMSCs.It also dose-dependently inhibited lipid droplet formation and decreased the expression of PPARG and FABP4 in NSAA-derived BMSCs.NFATC4 expression was higher in patients with NSAA than in healthy controls,and SJD downregulated its expression.NFATC4 overexpression reversed the inhibitory effect of SJD on adipogenic differentiation.Additionally,SJD promoted the deacetylation of PGC-1αin NSAA-derived BMSCs,which was also partially eliminated by NFATC4 overexpression.CONCLUSIONS:SJD inhibits adipogenic differentiation of BMSCs through downregulating NFATC4,thereby contributing to the remission of NSAA.展开更多
YAP（yes-associated protein） is a transcriptional factor that is negatively regulated by Hippo pathway, a conserved pathway for the development and size control of multiple organs. The exact function of YAP in bone h...YAP（yes-associated protein） is a transcriptional factor that is negatively regulated by Hippo pathway, a conserved pathway for the development and size control of multiple organs. The exact function of YAP in bone homeostasis remains controversial. Here we provide evidence for YAP＇s function in promoting osteogenesis, suppressing adipogenesis, and thus maintaining bone homeostasis.YAP is selectively expressed in osteoblast（OB）-lineage cells. Conditionally knocking out Yap in the OB lineage in mice reduces cell proliferation and OB differentiation and increases adipocyte formation, resulting in a trabecular bone loss. Mechanistically, YAP interacts with β-catenin and is necessary for maintenance of nuclear β-catenin level and Wnt/β-catenin signaling. Expression of β-catenin in YAP-deficient BMSCs（bone marrow stromal cells） diminishes the osteogenesis deficit. These results thus identify YAP-β-catenin as an important pathway for osteogenesis during adult bone remodeling and uncover a mechanism underlying YAP regulation of bone homeostasis.展开更多
Mesenchymal stem cells(MSCs)are stem/progenitor cells capable of self-renewal and differentiation into osteoblasts,chondrocytes and adipocytes.The transformation of multipotent MSCs to adipocytes mainly involves two s...Mesenchymal stem cells(MSCs)are stem/progenitor cells capable of self-renewal and differentiation into osteoblasts,chondrocytes and adipocytes.The transformation of multipotent MSCs to adipocytes mainly involves two subsequent steps from MSCs to preadipocytes and further preadipocytes into adipocytes,in which the process MSCs are precisely controlled to commit to the adipogenic lineage and then mature into adipocytes.Previous studies have shown that the master transcription factors C/enhancer-binding protein alpha and peroxisome proliferation activator receptor gamma play vital roles in adipogenesis.However,the mechanism underlying the adipogenic differentiation of MSCs is not fully understood.Here,the current knowledge of adipogenic differentiation in MSCs is reviewed,focusing on signaling pathways,noncoding RNAs and epigenetic effects on DNA methylation and acetylation during MSC differentiation.Finally,the relationship between maladipogenic differentiation and diseases is briefly discussed.We hope that this review can broaden and deepen our understanding of how MSCs turn into adipocytes.展开更多
Objective:Polyploid giant cancer cells(PGCCs)with daughter cells express epithelial–mesenchymal transition(EMT)-associated proteins.Highly malignant tumor cells with EMT properties can transdifferentiate into mature ...Objective:Polyploid giant cancer cells(PGCCs)with daughter cells express epithelial–mesenchymal transition(EMT)-associated proteins.Highly malignant tumor cells with EMT properties can transdifferentiate into mature tumor cells.In this study,we elucidated the potential for,and underlying mechanism of,adipogenic differentiation of PGCCs with daughter cells(PDCs).Methods:Cobalt chloride was used to induce PGCC formation in HEY(wild-type P53)and MDA-MB-231(mutant P53)cells;these cells were then cultured in adipogenic differentiation medium.Oil red O staining was used to confirm adipogenic differentiation,and the cell cycle was detected with flow cytometry.The expression of adipogenic differentiation-associated proteins and P300 histone acetyltransferase activity were compared before and after adipogenic differentiation.Animal xenograft models were used to confirm the adipogenic differentiation of PDCs.Results:PDCs transdifferentiated into functional adipocytes.Two different cell cycle distributions were observed in PDCs after adipogenic differentiation.The expression levels of PPARγ,Ace-PPARγ,and Ace-P53 were higher in PDCs after adipogenic differentiation than in cells before adipogenic differentiation.Ace-PPARγand FABP4 expression increased in HEY cells and decreased in MDA-MB-231 PDCs after p53 knockdown.A485 treatment increased Ace-P53,Ace-PPARγ,and FABP4 expression in HEY PDCs by inhibiting SUMOylation of P53.In MDA-MB-231 PDCs,A485 treatment decreased Ace-P53,Ace-PPARγ,and FABP4 expression.Animal experiments also confirmed the adipogenic differentiation of PDCs.Conclusions:Acetylation of P53 and PPARγplays an important role in the adipogenic differentiation of PDCs.展开更多
In order to elucidate the action of La3＋ on bone metabolism,effects of La3＋ on the osteogenic and adipogenic differentiation of pri-mary mouse bone marrow stromal cells（BMSCs） were studied by 3-（4,5-dimethylthiaz...In order to elucidate the action of La3＋ on bone metabolism,effects of La3＋ on the osteogenic and adipogenic differentiation of pri-mary mouse bone marrow stromal cells（BMSCs） were studied by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide（MTT） test,alkaline phosphatase（ALP） activity measurement,mineralized function,oil red O stain and measurement.The results showed that La3＋ pro-moted the proliferation of BMSCs except at 1×10-10 and 1×10-6 mol/L.The effect of La3＋ on the osteogenic differentiation depended on con-centrations at the 7th day,but the osteogenic differentiation was inhibited at any concentration at the 14th day.La3＋ promoted the formation of mineralized matrix nodules except at 1×10-8 and 1×10-5 mol/L.La3＋ inhibited adipogenic differentiation except at 1×10-10 and 1×10-7 mol/L at the 10th day,and inhibited adipogenic differentiation except at 1×10-9 mol/L at the 16th day.These findings suggested that La3＋ might have protective effect on bone at appropriate dose and time.This would be valuable for better understanding the mechanism of the effect of La3＋ on bone metabolism.展开更多
AIM: To study the metabolic profile of human umbilical mesenchymal stem cells (HUMSC) and adipogenic differentiation by nuclear magnetic resonance (NMR) spectroscopy.
Mesenchymal stem cells(MSCs)are multi-potent cells that are able to differentiate and mature into various types of cells under a certain microenvironment for cell therapy and tissue regeneration.Scandium(Sc),an import...Mesenchymal stem cells(MSCs)are multi-potent cells that are able to differentiate and mature into various types of cells under a certain microenvironment for cell therapy and tissue regeneration.Scandium(Sc),an important rare earth element,recently has been intensively investigated in biomedical fields as well as industrial engineering,and chloride channels have been proven to be able to affect osteogenic differentiation.Thus,it is significant to investigate effects of ScCl_(3)on cell activities of MSCs.In this paper,rat bone MSCs(rBMSCs)were co-cultured with various concentrations of ScCl_(3)(1×10^(-8),1×10^(-6),and 1×10^(-4)mol/L)to evaluate their influence on cell proliferation as well as osteogenic and adipogenic differentiation in vitro.The results indicate that ScCl_(3)promotes the proliferation of rBMSCs initially,which is yet reduced upon ion accumulation.We used immunofluorescence staining,quantitative real time polymerase chain reactions,and assays measuring alkaline phosphatase activity,mineralized deposits,and intracytoplasmic lipids to reveal that rBMSCs treated with ScCl_(3)at concentrations of 1×10^(-8)-1×10^(-6)mol/L can enhance levels of osteogenic differentiation in a dosedependent manner and reduce adipogenic differentiation to a certain degree through Wnt/β-catenin signaling pathway.These results indicate that appropriate concentrations of ScCl_(3)can improve osteogenic differentiation in the lineage commitment of rBMSCs,and thus,promote bone remodeling.This study implies that ScCl_(3) possesses great potentials in the treatment of bone diseases and would provide new strategy of designing composites by SiCl3 doping for biomedical applications in the future.展开更多
Obesity is a major human health problem associated with various diseases, including cardiac injury and type 2 diabetes. Trapa japonica Flerov (TJF) has been used in traditional oriental medicine to treat diabetes. In ...Obesity is a major human health problem associated with various diseases, including cardiac injury and type 2 diabetes. Trapa japonica Flerov (TJF) has been used in traditional oriental medicine to treat diabetes. In this study, we evaluated the inhibitory effect of and the mechanism underlying the effect of TJF extract on adipogenesis in 3T3-L1 cells. The effects of TJF extract on cell viability were analyzed using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and the anti-adipogenic effect was measured by oil red O staining. The expression of peroxisomal proliferator activated receptor (PPAR)γ, CCAAT/enhancer-binding protein-α (C/EBP)α, adenosine monophosphate-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), adiponectin, and fatty acid binding protein (FABP)4 involved in adipogenesis was determined by western blot analysis. TJF extract effectively inhibited lipid accumulation and the expression of PPARγ and C/EBPα in 3T3-L1 cells. TJF also increased the phosphorylation of AMPK and ACC, and decreased the expression of adiponectin and FABP4. These results indicate that TJF extract exerts its anti-obesity effect through the downregulation of adipogenic transcription factors and adipogenic marker genes.展开更多
Objective:To determine the effect of Salvianolic acid B (Sal B) on glucose and lipid metabolism in mice with high-fat diet (HFD)-induced obesity,and to investigate the underlying mechanisms by measuring the expression...Objective:To determine the effect of Salvianolic acid B (Sal B) on glucose and lipid metabolism in mice with high-fat diet (HFD)-induced obesity,and to investigate the underlying mechanisms by measuring the expression levels of key adipogenic transcription factors.Methods:Six-week-old C57BL/6J male mice were fed for 12 weeks with a HFD to induce obesity or a standard diet to serve as normal controls.A mean body weight increase of more than 20% after these 12 weeks was used as the criteria for obesity.HFD-fed obese mice then received a supplement of Sal B (100 mg/kg body weight/day),metformin (75 mg/kg body weight/day) or water (an equivalent volume;served as model controls) by oral gavage for an additional 8 weeks,and the normal controls received water (an equivalent volume) by oral gavage for the same period.Results:Sal B significantly reduced body weight gain (P <.05) without influencing food intake in HFD-fed obese mice relative to model controls.Sal B also reduced the body fat mass of the obese mice relative to model controls in a time-dependent manner (P <.05).Sal B significantly decreased the serum concentrations of low-density lipoprotein cholesterol,total cholesterol,triglyceride and free fatty acids by 25.5%,20.2%,20.6% and 13.4%,respectively,and increased the concentration of high-density lipoprotein cholesterol by 50.1% relative to model controls.In addition,Sal B significantly lowered fasting glucose concentrations and improved insulin sensitivity relative to model controls (P <.05).Sal B acted by ameliorating the histopathological changes in both brown and white adipose tissues of obese mice.Moreover,in brown adipose tissue,Sal B up-regulated the mRNA and protein expression of PPARγ and c/EBPα,and the protein expression of PPARα and SREBP-1 (P <.05).In white adipose tissue,Sal B down-regulated the mRNA expression of PPARγ and c/EBPα,and decreased the protein expression of PPARγ and SREBP-1(P <.05).Conclusjons:The results suggest that Sal B can reduce body weight gain and regulate glucose and lipid metabolism in mice with diet-induced obesity by regulating adipogenic transcription factors in their adipose tissues.展开更多
Imipramine （IM） has been widely used clinically for the treatment of mental disorders. Its actions on tissues or organs other than the nervous system also need to be understood for its proper clinical use. In this s...Imipramine （IM） has been widely used clinically for the treatment of mental disorders. Its actions on tissues or organs other than the nervous system also need to be understood for its proper clinical use. In this study, the effects of IM on adipogenic differentiation in both 3T3-L1 preadipocytes and mouse marrow stromal cells （MSCs） were investigated. The results showed that fewer adipocytic cells were developed from 3T3-L1 preadipocytes in the presence of 0.001 to 1 μmol/L of IM as compared to control. Similar inhibitory effect was also observed in mouse MSCs. The decrease in the formation of adipocytes was accompanied with significant down-regulation at mRNA expression of the early adipogenic transcription factor, peroxisome proliferator-activated receptor γ2 （PPARγ2）. Western blot analysis further revealed that the protein expression of PPARγ2 was reduced markedly in ceils treated with IM at concentrations of 0.01, 0.1 and 1 μmol/L, suggesting that the suppression on PPAR72 was involved in IM＇s inhibition on MSCs adipogenesis. Moreover, IM at the above concentrations could stimulate the mRNA expression of β2-adrenergic receptor （AR） and β3-AR, which implicated that the effect of IM on adipogenic differentiation was partially mediated by β-ARs. Our results demonstrated for the first time that the conventional antidepressive imipramine exerts accompanied inhibitory effect on adipocyte formation, which may have possible clinical implications.展开更多
Cultured meat provides a sustainable and safe alternative to traditional meat production by culturing animal cells in vitro.Cultured fat is an important component of cultured meat,contributing to its flavor,texture,an...Cultured meat provides a sustainable and safe alternative to traditional meat production by culturing animal cells in vitro.Cultured fat is an important component of cultured meat,contributing to its flavor,texture,and nutrition.Induced adipogenic differentiation is the most important step in the production of cultured fat,but conventional adipogenic inducer cocktails are complex and contain non-food-grade substances,which introduces a food safety risk for cultured meat products.Here we demonstrated that a food-grade substance,peanut oil(P-OIL),promoted adipogenic differentiation and lipid accumulation of bovine adipose-derived stem cells,based on which a simple and efficient approach was developed to produce cultured bovine fat.Mechanistic analysis showed that P-OIL upregulated genes involved in lipid synthesis and storage,such as peroxisome proliferator-activated receptor y(PPARy),carnitine palmitoyltransferase 2,and lipoprotein lipase,by activating the PPAR signaling pathway during adipogenesis.Notably,the lipid composition of cultured bovine fats generated using P-OIL induction was similar to that of fats obtained from farmed cattle,but free of trans-fatty acids.This study provides valuable insights into the production of safe,healthy,and nutritious cultured fats.展开更多
Background Steroids inhibit osteogenic differentiation and decrease bone formation while concomitantly inducing adipose deposition in osteocytes.This leads to the fatty degeneration and necrosis of bone cells commonly...Background Steroids inhibit osteogenic differentiation and decrease bone formation while concomitantly inducing adipose deposition in osteocytes.This leads to the fatty degeneration and necrosis of bone cells commonly seen in osteonecrosis of the femoral head.The peroxisome proliferator-activated receptor-γ （PPARγ） is an adipogenic transcription factor linked to the development of this disease and responsible for inducing adipogenesis over osteogenesis in bone marrow mesenchymal stem cells （BMSCs）.The aim of this study was to assess whether adipogenic differentiation could be suppressed,and thus osteogenic potential retained,by inhibiting PPARy expression in BMSCs.Methods Cells from the bone marrow of New Zealand rabbits were treated with 10-7 mol/L dexamethasone and infected with one of three small interference RNA （siRNA） adenovirus vectors （S1,S2,and S3） or non-targeting control siRNA （Con） and compared with dexamethasone-treated （model） and untreated （normal） cells.Cells were grown for 21 days and stained with Sudan Ⅲ for adipocyte formation.At various time points,cells were also assessed for changes in PPARγ,osteocalcin （OC）,Runx2,alkaline phosphatase （ALP） activity,and triglyceride （TG） content.Results Dexamethasone-treated model and control groups showed a significant increase in fatty acid-positive staining,which was inhibited in cells treated with PPARγ siRNA-treated,similar to normal untreated cells.All three siRNA groups significantly inhibited PPARγ mRNA and protein,adipocyte number,and TG content compared with the dexamethasonetreated model and control groups,matching that seen in normal cells.OC and Runx2 mRNA and protein,as well as ALP activity,were significantly higher in cells treated with siRNA against PPARγ,similar to that seen in the normal cells.These osteogenic markers were significantly lower in the dexamethasone-treated cell cultures.Conclusions The siRNA adenovirus vector targeting PPARγ can efficiently inhibit steroid-induced adipogenic differentiation in rabbit BMSCs and retain their osteogenic differentiation potential.展开更多
Porcine mesenchymal stem cells in postnatal muscle have been demonstrated to differentiate into adipocytes. This increases adipocyte number and lipid accumulation, and is thought to be the origin of intramuscular fat....Porcine mesenchymal stem cells in postnatal muscle have been demonstrated to differentiate into adipocytes. This increases adipocyte number and lipid accumulation, and is thought to be the origin of intramuscular fat. In this study, the effects of myostatin and arginine on adipogenic differentiation in mesenchymal stem cells derived from porcine muscle （pMDSCs） were investigated in vitro. Intracellular triglyceride levels were reduced by exogenous myostatin and increased by arginine supplementation or myostatin antibody （P〈0.01）. The inhibition of lipid accumulation by rnyostatin in pMDSCs was alleviated by arginine supplementation （P〈0.01）. Expression patterns of adipogenic transcription factors showed that exogenous myostatin suppressed PPAR72 and aP2 expression （P〈0.01）, while supplemental arginine or myostatin antibody promoted ADD1 expression （P〈0.01）. Furthermore, compared with the addition of either myostatin protein or antibody alone, ADD1 and PPARδ expression were promoted by the combination of arginine and myostatin （P〈0.01）, and arginine combined with myostatin antibody promoted the expression of ADD1, PPARδ, C/EBPα, PPARγ2 and LPL in pMDSCs （P〈0.05）. These results suggest that myostatin inhibits adipogenesis in pMDSCs, and that this can be alleviated by arginine supplementation, at least in part, through promoting ADD1 and PPARδ expression.展开更多
Extracellular matrix （ECM） plays a very important role in regulating cell function and fate. It is highly desirable to fabricate biomimetic models to investigate the role of ECM in stem cell differentiation. In this...Extracellular matrix （ECM） plays a very important role in regulating cell function and fate. It is highly desirable to fabricate biomimetic models to investigate the role of ECM in stem cell differentiation. In this study, arginine- glycine--aspartate （RGD）-modified gold nanoparticles （Au NPs） with tunable surface ligand density were prepared to mimic the ECM microenvironment. Their effect on osteogenic and adipogenic differentiation of human mesenchymal stem cells （MSCs） was investigated. The biomimetic Au NPs were taken up by MSCs in a ligand density-dependent manner. The biomimetic NPs with a high RGD density had an inhibitive effect on the alkaline phosphatase （ALP） activity, calcium deposition, and osteogenic marker gene expression of MSCs. Their effect on oil droplet formation and adipogenic marker gene expression was negative when RGD density was low, while their effect was promotive when RGD density was high. The biomimetic Au NPs regulated the osteogenic and adipogenic differentiation of MSCs mainly through affecting the focal adhesion and cytoskeleton. This study highlights the roles of biomimetic NPs on stem cell differentiation that could provide a meaningful strategy in fabricating functional biomaterials for tissue engineering and biomedical applications.展开更多
The retinoblastoma gene product(pRb)is a chromatin-associated protein that can either suppress or promote activity of key regulators of tissue-specific differentiation.We found that twelve weeks after transfection of ...The retinoblastoma gene product(pRb)is a chromatin-associated protein that can either suppress or promote activity of key regulators of tissue-specific differentiation.We found that twelve weeks after transfection of the exogenous active(ΔB/X andΔр34)or inactive(ΔS/N)forms of RB into the 10T1/2 mesenchymal stem cells and clonal selection not a single cell line did contain exogenous RB,despite being G-418 resistant.However,the consequences of the transient production of exogenous RB had different effects on the cell fate.TheΔB/X andΔр34 cells transfected with active form of RB showed elevated levels of inducible adipocyte differentiation(AD).On the contrary,theΔS/N cells transfected with inactive RB mutant were insensitive to induction of AD associated with abolishing of expression of the PPARγ2.Additionally,the PPARγ2 promoter in undifferentiatedΔS/N cells was hypermethylated,but all except−60 position CpG became mostly demethylated after cells exposure to AD.We conclude that while transient expression of inactive exogenous RB induces long term epigenetic alterations that prevent adipogenesis,production of active exogenous RBs results in an AD-promoting epigenetic state.These results indicate that pRb is involved in the establishment of hereditary epigenetic memory at least by creating a methylation pattern of PPARγ2.展开更多
Objective：The study was designed to investigate the molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of r BMSCs.Methods：r BMSCs were harvested from SD rats,and determina...Objective：The study was designed to investigate the molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of r BMSCs.Methods：r BMSCs were harvested from SD rats,and determination of alkaline phosphatase（ALP）activity,quantification of mineralization by Alizarin Red S staining,and the m RNA expression of osteogenic differentiation markers（Runx2,BMP-2,and OSX）by RT-PCR after r BMSCs stimulated by osteogenic induction with（0.1–10）μg/m L of quercitrin,quantification of Lipid droplet by Oil Red O staining and the m RNA expression of adipogenic differentiation marker（,and a P2）by RT-PCR after r BMSCs stimulated by adipogenic induction with（0.1-10）μg/m L of quercitrin.Results：Quercitrin can up-regulate the m RNA expression of osteogenic differentiation markers（Runx2,BMP-2,and OSX）and increase ALP activity and mineralization after osteogenic induction,on the other hand quercitrin can suppress the m RNA expression of adipogenic differentiation markers（,and a P2）and decrease lipid droplet after adipogenic induction.Conclusion：This study suggested that quercitrin not only stimulated osteogenic differentiation but also inhibited adipogenic differentiation of r BMSCs,which was associated with the up-regulation of Runx2,BMP-2,and OSX m RNA expression and the down-regulation of,and a P2 m RNA expression.展开更多
Aim:To evaluate the age-related effects on the adipogenic differentiation and proliferation potentials of human orbital adipose-derived stem cells(OASCs).Methods:Orbital adipose samples were harvested from the central...Aim:To evaluate the age-related effects on the adipogenic differentiation and proliferation potentials of human orbital adipose-derived stem cells(OASCs).Methods:Orbital adipose samples were harvested from the central fat compartment in the lower eyelids of 10 young and middle-aged patients during routine blepharoplasty surgery.After assessment of the morphological changes of adipocytes with aging,OASCs were isolated from the fat samples and expanded in vitro.Differences in the stem cell colony number(fibroblast colony-forming unit),growth rate and phenotype characterization(flow cytometry analysis)were evaluated.The ability of OASCs to differentiate into adipocytes was determined by oil red O staining and the mRNA expression level of peroxisome proliferator-activated receptorγ.Results:Fat cell size showed a decreasing trend with advancing age.Although no difference was found in the expression of cell surface markers,the colony number and proliferative rate of OASCs from middle-aged donors were significantly lower than those from the young donors.The adipogenic differentiation capacity of middle-aged OASCs was also reduced.These differences were statistically significant(P<0.001).Conclusion:The data showed that the progenitor cell number,proliferation capacity and adipogenic potential of OASCs decreased with aging,suggesting that using OASCs from elderly patients for therapeutic purposes might be restricted.展开更多
During excessive adipose tissue accumulation,various adipokines such as visfatin,chemerin,vaspin,and adiponectin are released into systemic circulation,thereby influencing metabolic tissue function throughout the body...During excessive adipose tissue accumulation,various adipokines such as visfatin,chemerin,vaspin,and adiponectin are released into systemic circulation,thereby influencing metabolic tissue function throughout the body.As multifunctional signaling molecules secreted by adipose tissue,adipokines play a pivotal role in metabolic regulation,inflammatory response,and tissue homeostasis.Recent studies have demonstrated that adipokines can influence skeletal system repair and regeneration by modulating bone marrow-derived mesenchymal stem cell(BMSC)proliferation,differentiation,migration,and immunomodulatory functions.However,different adipokines have distinct roles in regulating BMSC function,but their underlying molecular mechanisms are not fully understood.In this review,we systematically summarize the specific mechanisms of action and potential clinical applications of visfatin,chemerin,vaspin,and adiponectin on BMSC function in order to reveal new mechanisms of interaction between adipokines and BMSCs.The aim is to provide a theoretical basis for targeted treatment strategies for bone diseases targeting adipokines.展开更多
Background:Long non-coding RNAs(lncRNAs)are emerging key regulators involved in a variety of biological processes such as cell differentiation and development.The balance between myogenesis and adipogenesis is crucial...Background:Long non-coding RNAs(lncRNAs)are emerging key regulators involved in a variety of biological processes such as cell differentiation and development.The balance between myogenesis and adipogenesis is crucial for skeletal muscle homeostasis in humans and meat quality in farm animals.The present study aimed to reveal the global transcriptomic profiles of adipogenic(Adi-)and myogenic(Myo-)precursors derived from porcine skeletal muscle and identify lncRNAs involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Results:In this study,a total of 655 novel individual lncRNAs including differentially expressed 24 lncRNAs,and 755 differentially expressed mRNAs were identified(fold change≥2 or≤0.5 and adjusted P<0.05).Integrated results of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis accompanied by the variation of intracellular Ca2+concentration highlighted Lnc-ADAMTS9 involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Although Lnc-ADAMTS9 knock-down did not alter the mRNA expression of ADAMTS9,we demonstrated that Lnc-ADAMTS9 can promote myogenic proliferation and myogenic differentiation of myogenic precursors through inhibiting the ERK/MAPK signaling pathway.Conclusion:We deciphered a comprehensive catalog of mRNAs and lncRNAs that might be involved in the regulation of myogenesis and adipogenesis homeostasis in the skeletal muscle of pigs.The Lnc-ADAMTS9 exerts an essential role in myogenesis through the ERK signaling pathway.展开更多
基金supported by NIH grants NIH/NIA R01AG069401(to L.Q.)NIH/NHLBI U54HL165442(to K.T.)P30AR069619(to Penn Center for Musculoskeletal Disorders).
文摘Bone resorption by osteoclasts is a critical step in bone remodeling,a process important for maintaining bone homeostasis and repairing injured bone.We previously identified a bone marrow mesenchymal subpopulation,marrow adipogenic lineage precursors(MALPs),and showed that its production of RANKL stimulates bone resorption in young mice using Adipoq-Cre.To exclude developmental defects and to investigate the role of MALPs-derived RANKL in adult bone,we generated inducible reporter mice(Adipoq-CreER Tomato)and RANKL deficient mice(Adipoq-CreER RANKLflox/flox,iCKO).Single cell-RNA sequencing data analysis and lineage tracing revealed that Adipoq+cells contain not only MALPs but also some mesenchymal progenitors capable of osteogenic differentiation.In situ hybridization showed that RANKL mRNA is only detected in MALPs,but not in osteogenic cells.RANKL deficiency in MALPs induced at 3 months of age rapidly increased trabecular bone mass in long bones as well as vertebrae due to diminished bone resorption but had no effect on the cortical bone.Ovariectomy(OVX)induced trabecular bone loss at both sites.RANKL depletion either before OVX or at 6 weeks post OVX protected and restored trabecular bone mass.Furthermore,bone healing after drill-hole injury was delayed in iCKO mice.Together,our findings demonstrate that MALPs play a dominant role in controlling trabecular bone resorption and that RANKL from MALPs is essential for trabecular bone turnover in adult bone homeostasis,postmenopausal bone loss,and injury repair.
基金Supported by Zhejiang Province Traditional Chinese Medicine Science and Technology Plan Project:Research on the Mechanism of Shisiwei Jianzhong Decoction in Treating Non-severe Aplastic Anemia with Kidney Yang Deficiency Based on the C Vactivated T Nuclear Factor(NFATc4)(2020ZB086)Zhejiang Province Traditional Chinese Medicine Science and Technology Youth Talent Plan Project:Study on the Academic thought and Clinical Application of Professor Zhou Yuhong in the Treatment of Chronic Aplastic Anemia(2021ZQ029)。
文摘OBJECTIVE:To investigate the effect of Shisiwei Jianzhong decoction(十四味建中汤,SJD)on non-severe aplastic anemia(NSAA).METHODS:Bone marrow mesenchymal stem cells(BMSCs)were isolated from bone marrow samples of 15 NSAA patients and 3 healthy controls.Cells were treated with gradient concentrations of SJD,and a portion was transfected with a vector overexpressing the nuclear factor of activated T cells,cytoplasmic 4(NFATC4).Cell viability and apoptosis were detected by cell counting kit-8 and flow cytometry,respectively.After adipogenic differentiation induction,lipid droplet formation in BMSCs was examined by Oil Red O staining.The expression of NFATC4,peroxisome proliferator-activated receptor gamma(PPARG),fatty acid-binding protein 4(FABP4),peroxisome proliferator-activated receptor-gamma coactivator(PGC-1α),and acetylated PGC-1αwas measured by quantitative real-time polymerase chain reaction or Western blot.RESULTS:SJD significantly increased the viability and decreased the apoptosis of NSAA-derived BMSCs.It also dose-dependently inhibited lipid droplet formation and decreased the expression of PPARG and FABP4 in NSAA-derived BMSCs.NFATC4 expression was higher in patients with NSAA than in healthy controls,and SJD downregulated its expression.NFATC4 overexpression reversed the inhibitory effect of SJD on adipogenic differentiation.Additionally,SJD promoted the deacetylation of PGC-1αin NSAA-derived BMSCs,which was also partially eliminated by NFATC4 overexpression.CONCLUSIONS:SJD inhibits adipogenic differentiation of BMSCs through downregulating NFATC4,thereby contributing to the remission of NSAA.
基金supported in part by grants from the National Institutes of Health(AG051773)and VA(BX000838)
文摘YAP（yes-associated protein） is a transcriptional factor that is negatively regulated by Hippo pathway, a conserved pathway for the development and size control of multiple organs. The exact function of YAP in bone homeostasis remains controversial. Here we provide evidence for YAP＇s function in promoting osteogenesis, suppressing adipogenesis, and thus maintaining bone homeostasis.YAP is selectively expressed in osteoblast（OB）-lineage cells. Conditionally knocking out Yap in the OB lineage in mice reduces cell proliferation and OB differentiation and increases adipocyte formation, resulting in a trabecular bone loss. Mechanistically, YAP interacts with β-catenin and is necessary for maintenance of nuclear β-catenin level and Wnt/β-catenin signaling. Expression of β-catenin in YAP-deficient BMSCs（bone marrow stromal cells） diminishes the osteogenesis deficit. These results thus identify YAP-β-catenin as an important pathway for osteogenesis during adult bone remodeling and uncover a mechanism underlying YAP regulation of bone homeostasis.
基金Supported by the National Natural Science Foundation of China,No.82271843 and 31700779the Key Project supported by Medical Science and Technology Development Foundation,Nanjing Department of Health,No.ZKX20019the Natural Science Foundation of Jiangsu Province,No.BK20200137.
文摘Mesenchymal stem cells(MSCs)are stem/progenitor cells capable of self-renewal and differentiation into osteoblasts,chondrocytes and adipocytes.The transformation of multipotent MSCs to adipocytes mainly involves two subsequent steps from MSCs to preadipocytes and further preadipocytes into adipocytes,in which the process MSCs are precisely controlled to commit to the adipogenic lineage and then mature into adipocytes.Previous studies have shown that the master transcription factors C/enhancer-binding protein alpha and peroxisome proliferation activator receptor gamma play vital roles in adipogenesis.However,the mechanism underlying the adipogenic differentiation of MSCs is not fully understood.Here,the current knowledge of adipogenic differentiation in MSCs is reviewed,focusing on signaling pathways,noncoding RNAs and epigenetic effects on DNA methylation and acetylation during MSC differentiation.Finally,the relationship between maladipogenic differentiation and diseases is briefly discussed.We hope that this review can broaden and deepen our understanding of how MSCs turn into adipocytes.
基金supported partly by grants from the National Natural Science Foundation of China(Grant Nos.82173283 and 82103088)the Foundation of the Committee on Science and Technology of Tianjin(Grant No.20JCYBJC01230)。
文摘Objective:Polyploid giant cancer cells(PGCCs)with daughter cells express epithelial–mesenchymal transition(EMT)-associated proteins.Highly malignant tumor cells with EMT properties can transdifferentiate into mature tumor cells.In this study,we elucidated the potential for,and underlying mechanism of,adipogenic differentiation of PGCCs with daughter cells(PDCs).Methods:Cobalt chloride was used to induce PGCC formation in HEY(wild-type P53)and MDA-MB-231(mutant P53)cells;these cells were then cultured in adipogenic differentiation medium.Oil red O staining was used to confirm adipogenic differentiation,and the cell cycle was detected with flow cytometry.The expression of adipogenic differentiation-associated proteins and P300 histone acetyltransferase activity were compared before and after adipogenic differentiation.Animal xenograft models were used to confirm the adipogenic differentiation of PDCs.Results:PDCs transdifferentiated into functional adipocytes.Two different cell cycle distributions were observed in PDCs after adipogenic differentiation.The expression levels of PPARγ,Ace-PPARγ,and Ace-P53 were higher in PDCs after adipogenic differentiation than in cells before adipogenic differentiation.Ace-PPARγand FABP4 expression increased in HEY cells and decreased in MDA-MB-231 PDCs after p53 knockdown.A485 treatment increased Ace-P53,Ace-PPARγ,and FABP4 expression in HEY PDCs by inhibiting SUMOylation of P53.In MDA-MB-231 PDCs,A485 treatment decreased Ace-P53,Ace-PPARγ,and FABP4 expression.Animal experiments also confirmed the adipogenic differentiation of PDCs.Conclusions:Acetylation of P53 and PPARγplays an important role in the adipogenic differentiation of PDCs.
基金Project supported by the National Natural Science Foundation of China (20971034)Foundation for Key Program of Ministry of Education of China (208018)+2 种基金Returned Scholars of Hebei Province (207041)Natural Science Key Foundation of Hebei Province (B2009000161)Natural Science Foundation of Hebei University
文摘In order to elucidate the action of La3＋ on bone metabolism,effects of La3＋ on the osteogenic and adipogenic differentiation of pri-mary mouse bone marrow stromal cells（BMSCs） were studied by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide（MTT） test,alkaline phosphatase（ALP） activity measurement,mineralized function,oil red O stain and measurement.The results showed that La3＋ pro-moted the proliferation of BMSCs except at 1×10-10 and 1×10-6 mol/L.The effect of La3＋ on the osteogenic differentiation depended on con-centrations at the 7th day,but the osteogenic differentiation was inhibited at any concentration at the 14th day.La3＋ promoted the formation of mineralized matrix nodules except at 1×10-8 and 1×10-5 mol/L.La3＋ inhibited adipogenic differentiation except at 1×10-10 and 1×10-7 mol/L at the 10th day,and inhibited adipogenic differentiation except at 1×10-9 mol/L at the 16th day.These findings suggested that La3＋ might have protective effect on bone at appropriate dose and time.This would be valuable for better understanding the mechanism of the effect of La3＋ on bone metabolism.
基金Supported by Grants from the National Natural Science Foundation of China(Key program 30930027)Natural Science Foundation of Guangdong Province(No.8151503102000032)
文摘AIM: To study the metabolic profile of human umbilical mesenchymal stem cells (HUMSC) and adipogenic differentiation by nuclear magnetic resonance (NMR) spectroscopy.
基金Project supported by the National Natural Science Foundation of China(51972148,51802115,11904131,21501090)the Project of"20 items of University"of Jinan(2018GXRC031)the Doctoral Fund of University of Jinan(XBS1609)。
文摘Mesenchymal stem cells(MSCs)are multi-potent cells that are able to differentiate and mature into various types of cells under a certain microenvironment for cell therapy and tissue regeneration.Scandium(Sc),an important rare earth element,recently has been intensively investigated in biomedical fields as well as industrial engineering,and chloride channels have been proven to be able to affect osteogenic differentiation.Thus,it is significant to investigate effects of ScCl_(3)on cell activities of MSCs.In this paper,rat bone MSCs(rBMSCs)were co-cultured with various concentrations of ScCl_(3)(1×10^(-8),1×10^(-6),and 1×10^(-4)mol/L)to evaluate their influence on cell proliferation as well as osteogenic and adipogenic differentiation in vitro.The results indicate that ScCl_(3)promotes the proliferation of rBMSCs initially,which is yet reduced upon ion accumulation.We used immunofluorescence staining,quantitative real time polymerase chain reactions,and assays measuring alkaline phosphatase activity,mineralized deposits,and intracytoplasmic lipids to reveal that rBMSCs treated with ScCl_(3)at concentrations of 1×10^(-8)-1×10^(-6)mol/L can enhance levels of osteogenic differentiation in a dosedependent manner and reduce adipogenic differentiation to a certain degree through Wnt/β-catenin signaling pathway.These results indicate that appropriate concentrations of ScCl_(3)can improve osteogenic differentiation in the lineage commitment of rBMSCs,and thus,promote bone remodeling.This study implies that ScCl_(3) possesses great potentials in the treatment of bone diseases and would provide new strategy of designing composites by SiCl3 doping for biomedical applications in the future.
文摘Obesity is a major human health problem associated with various diseases, including cardiac injury and type 2 diabetes. Trapa japonica Flerov (TJF) has been used in traditional oriental medicine to treat diabetes. In this study, we evaluated the inhibitory effect of and the mechanism underlying the effect of TJF extract on adipogenesis in 3T3-L1 cells. The effects of TJF extract on cell viability were analyzed using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and the anti-adipogenic effect was measured by oil red O staining. The expression of peroxisomal proliferator activated receptor (PPAR)γ, CCAAT/enhancer-binding protein-α (C/EBP)α, adenosine monophosphate-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), adiponectin, and fatty acid binding protein (FABP)4 involved in adipogenesis was determined by western blot analysis. TJF extract effectively inhibited lipid accumulation and the expression of PPARγ and C/EBPα in 3T3-L1 cells. TJF also increased the phosphorylation of AMPK and ACC, and decreased the expression of adiponectin and FABP4. These results indicate that TJF extract exerts its anti-obesity effect through the downregulation of adipogenic transcription factors and adipogenic marker genes.
基金This study is supported by grants from the National Natural Science Foundation of China(81274041 and 81503540)the International Cooperation Projects of MOE(2011DFA30920)+1 种基金a Co-construction Project of Beijing Board of Education(0101216-14)a Research Project of the Beijing University of Chinese Medicine(2014-X-003).
文摘Objective:To determine the effect of Salvianolic acid B (Sal B) on glucose and lipid metabolism in mice with high-fat diet (HFD)-induced obesity,and to investigate the underlying mechanisms by measuring the expression levels of key adipogenic transcription factors.Methods:Six-week-old C57BL/6J male mice were fed for 12 weeks with a HFD to induce obesity or a standard diet to serve as normal controls.A mean body weight increase of more than 20% after these 12 weeks was used as the criteria for obesity.HFD-fed obese mice then received a supplement of Sal B (100 mg/kg body weight/day),metformin (75 mg/kg body weight/day) or water (an equivalent volume;served as model controls) by oral gavage for an additional 8 weeks,and the normal controls received water (an equivalent volume) by oral gavage for the same period.Results:Sal B significantly reduced body weight gain (P <.05) without influencing food intake in HFD-fed obese mice relative to model controls.Sal B also reduced the body fat mass of the obese mice relative to model controls in a time-dependent manner (P <.05).Sal B significantly decreased the serum concentrations of low-density lipoprotein cholesterol,total cholesterol,triglyceride and free fatty acids by 25.5%,20.2%,20.6% and 13.4%,respectively,and increased the concentration of high-density lipoprotein cholesterol by 50.1% relative to model controls.In addition,Sal B significantly lowered fasting glucose concentrations and improved insulin sensitivity relative to model controls (P <.05).Sal B acted by ameliorating the histopathological changes in both brown and white adipose tissues of obese mice.Moreover,in brown adipose tissue,Sal B up-regulated the mRNA and protein expression of PPARγ and c/EBPα,and the protein expression of PPARα and SREBP-1 (P <.05).In white adipose tissue,Sal B down-regulated the mRNA expression of PPARγ and c/EBPα,and decreased the protein expression of PPARγ and SREBP-1(P <.05).Conclusjons:The results suggest that Sal B can reduce body weight gain and regulate glucose and lipid metabolism in mice with diet-induced obesity by regulating adipogenic transcription factors in their adipose tissues.
基金supported by a grant from UGC Area of Excellence project "Chinese Medicine Research and Further Development"(No.AoE/B-10/01)the Shenzhen Key Laboratory Funding Scheme of Shenzhen Municipal Government, Shenzhen Double Hundred Project
文摘Imipramine （IM） has been widely used clinically for the treatment of mental disorders. Its actions on tissues or organs other than the nervous system also need to be understood for its proper clinical use. In this study, the effects of IM on adipogenic differentiation in both 3T3-L1 preadipocytes and mouse marrow stromal cells （MSCs） were investigated. The results showed that fewer adipocytic cells were developed from 3T3-L1 preadipocytes in the presence of 0.001 to 1 μmol/L of IM as compared to control. Similar inhibitory effect was also observed in mouse MSCs. The decrease in the formation of adipocytes was accompanied with significant down-regulation at mRNA expression of the early adipogenic transcription factor, peroxisome proliferator-activated receptor γ2 （PPARγ2）. Western blot analysis further revealed that the protein expression of PPARγ2 was reduced markedly in ceils treated with IM at concentrations of 0.01, 0.1 and 1 μmol/L, suggesting that the suppression on PPAR72 was involved in IM＇s inhibition on MSCs adipogenesis. Moreover, IM at the above concentrations could stimulate the mRNA expression of β2-adrenergic receptor （AR） and β3-AR, which implicated that the effect of IM on adipogenic differentiation was partially mediated by β-ARs. Our results demonstrated for the first time that the conventional antidepressive imipramine exerts accompanied inhibitory effect on adipocyte formation, which may have possible clinical implications.
基金funded by the Agricultural Science and Technology Major Projectthe Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study(SN-ZJU-SIAS-0013)the Jiangsu Basic Research Center for Synthetic Biology(BK20233003)。
文摘Cultured meat provides a sustainable and safe alternative to traditional meat production by culturing animal cells in vitro.Cultured fat is an important component of cultured meat,contributing to its flavor,texture,and nutrition.Induced adipogenic differentiation is the most important step in the production of cultured fat,but conventional adipogenic inducer cocktails are complex and contain non-food-grade substances,which introduces a food safety risk for cultured meat products.Here we demonstrated that a food-grade substance,peanut oil(P-OIL),promoted adipogenic differentiation and lipid accumulation of bovine adipose-derived stem cells,based on which a simple and efficient approach was developed to produce cultured bovine fat.Mechanistic analysis showed that P-OIL upregulated genes involved in lipid synthesis and storage,such as peroxisome proliferator-activated receptor y(PPARy),carnitine palmitoyltransferase 2,and lipoprotein lipase,by activating the PPAR signaling pathway during adipogenesis.Notably,the lipid composition of cultured bovine fats generated using P-OIL induction was similar to that of fats obtained from farmed cattle,but free of trans-fatty acids.This study provides valuable insights into the production of safe,healthy,and nutritious cultured fats.
基金This study was supported by a grant from the National Natural Science Foundation of China （No. 81071520）.
文摘Background Steroids inhibit osteogenic differentiation and decrease bone formation while concomitantly inducing adipose deposition in osteocytes.This leads to the fatty degeneration and necrosis of bone cells commonly seen in osteonecrosis of the femoral head.The peroxisome proliferator-activated receptor-γ （PPARγ） is an adipogenic transcription factor linked to the development of this disease and responsible for inducing adipogenesis over osteogenesis in bone marrow mesenchymal stem cells （BMSCs）.The aim of this study was to assess whether adipogenic differentiation could be suppressed,and thus osteogenic potential retained,by inhibiting PPARy expression in BMSCs.Methods Cells from the bone marrow of New Zealand rabbits were treated with 10-7 mol/L dexamethasone and infected with one of three small interference RNA （siRNA） adenovirus vectors （S1,S2,and S3） or non-targeting control siRNA （Con） and compared with dexamethasone-treated （model） and untreated （normal） cells.Cells were grown for 21 days and stained with Sudan Ⅲ for adipocyte formation.At various time points,cells were also assessed for changes in PPARγ,osteocalcin （OC）,Runx2,alkaline phosphatase （ALP） activity,and triglyceride （TG） content.Results Dexamethasone-treated model and control groups showed a significant increase in fatty acid-positive staining,which was inhibited in cells treated with PPARγ siRNA-treated,similar to normal untreated cells.All three siRNA groups significantly inhibited PPARγ mRNA and protein,adipocyte number,and TG content compared with the dexamethasonetreated model and control groups,matching that seen in normal cells.OC and Runx2 mRNA and protein,as well as ALP activity,were significantly higher in cells treated with siRNA against PPARγ,similar to that seen in the normal cells.These osteogenic markers were significantly lower in the dexamethasone-treated cell cultures.Conclusions The siRNA adenovirus vector targeting PPARγ can efficiently inhibit steroid-induced adipogenic differentiation in rabbit BMSCs and retain their osteogenic differentiation potential.
基金supported by the National Natural Science Foundation of China (Grant No.30972119)
文摘Porcine mesenchymal stem cells in postnatal muscle have been demonstrated to differentiate into adipocytes. This increases adipocyte number and lipid accumulation, and is thought to be the origin of intramuscular fat. In this study, the effects of myostatin and arginine on adipogenic differentiation in mesenchymal stem cells derived from porcine muscle （pMDSCs） were investigated in vitro. Intracellular triglyceride levels were reduced by exogenous myostatin and increased by arginine supplementation or myostatin antibody （P〈0.01）. The inhibition of lipid accumulation by rnyostatin in pMDSCs was alleviated by arginine supplementation （P〈0.01）. Expression patterns of adipogenic transcription factors showed that exogenous myostatin suppressed PPAR72 and aP2 expression （P〈0.01）, while supplemental arginine or myostatin antibody promoted ADD1 expression （P〈0.01）. Furthermore, compared with the addition of either myostatin protein or antibody alone, ADD1 and PPARδ expression were promoted by the combination of arginine and myostatin （P〈0.01）, and arginine combined with myostatin antibody promoted the expression of ADD1, PPARδ, C/EBPα, PPARγ2 and LPL in pMDSCs （P〈0.05）. These results suggest that myostatin inhibits adipogenesis in pMDSCs, and that this can be alleviated by arginine supplementation, at least in part, through promoting ADD1 and PPARδ expression.
文摘Extracellular matrix （ECM） plays a very important role in regulating cell function and fate. It is highly desirable to fabricate biomimetic models to investigate the role of ECM in stem cell differentiation. In this study, arginine- glycine--aspartate （RGD）-modified gold nanoparticles （Au NPs） with tunable surface ligand density were prepared to mimic the ECM microenvironment. Their effect on osteogenic and adipogenic differentiation of human mesenchymal stem cells （MSCs） was investigated. The biomimetic Au NPs were taken up by MSCs in a ligand density-dependent manner. The biomimetic NPs with a high RGD density had an inhibitive effect on the alkaline phosphatase （ALP） activity, calcium deposition, and osteogenic marker gene expression of MSCs. Their effect on oil droplet formation and adipogenic marker gene expression was negative when RGD density was low, while their effect was promotive when RGD density was high. The biomimetic Au NPs regulated the osteogenic and adipogenic differentiation of MSCs mainly through affecting the focal adhesion and cytoskeleton. This study highlights the roles of biomimetic NPs on stem cell differentiation that could provide a meaningful strategy in fabricating functional biomaterials for tissue engineering and biomedical applications.
文摘The retinoblastoma gene product(pRb)is a chromatin-associated protein that can either suppress or promote activity of key regulators of tissue-specific differentiation.We found that twelve weeks after transfection of the exogenous active(ΔB/X andΔр34)or inactive(ΔS/N)forms of RB into the 10T1/2 mesenchymal stem cells and clonal selection not a single cell line did contain exogenous RB,despite being G-418 resistant.However,the consequences of the transient production of exogenous RB had different effects on the cell fate.TheΔB/X andΔр34 cells transfected with active form of RB showed elevated levels of inducible adipocyte differentiation(AD).On the contrary,theΔS/N cells transfected with inactive RB mutant were insensitive to induction of AD associated with abolishing of expression of the PPARγ2.Additionally,the PPARγ2 promoter in undifferentiatedΔS/N cells was hypermethylated,but all except−60 position CpG became mostly demethylated after cells exposure to AD.We conclude that while transient expression of inactive exogenous RB induces long term epigenetic alterations that prevent adipogenesis,production of active exogenous RBs results in an AD-promoting epigenetic state.These results indicate that pRb is involved in the establishment of hereditary epigenetic memory at least by creating a methylation pattern of PPARγ2.
基金supported by National Natural Science Foundation of China(No.81160508)The Science and Technology Landing Program Project of Colleges and University in Jiangxi Province(KJLD14058)
文摘Objective：The study was designed to investigate the molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of r BMSCs.Methods：r BMSCs were harvested from SD rats,and determination of alkaline phosphatase（ALP）activity,quantification of mineralization by Alizarin Red S staining,and the m RNA expression of osteogenic differentiation markers（Runx2,BMP-2,and OSX）by RT-PCR after r BMSCs stimulated by osteogenic induction with（0.1–10）μg/m L of quercitrin,quantification of Lipid droplet by Oil Red O staining and the m RNA expression of adipogenic differentiation marker（,and a P2）by RT-PCR after r BMSCs stimulated by adipogenic induction with（0.1-10）μg/m L of quercitrin.Results：Quercitrin can up-regulate the m RNA expression of osteogenic differentiation markers（Runx2,BMP-2,and OSX）and increase ALP activity and mineralization after osteogenic induction,on the other hand quercitrin can suppress the m RNA expression of adipogenic differentiation markers（,and a P2）and decrease lipid droplet after adipogenic induction.Conclusion：This study suggested that quercitrin not only stimulated osteogenic differentiation but also inhibited adipogenic differentiation of r BMSCs,which was associated with the up-regulation of Runx2,BMP-2,and OSX m RNA expression and the down-regulation of,and a P2 m RNA expression.
基金supported by the grants of National Natural Science Foundation of China(No.31271027 and No.81171475).
文摘Aim:To evaluate the age-related effects on the adipogenic differentiation and proliferation potentials of human orbital adipose-derived stem cells(OASCs).Methods:Orbital adipose samples were harvested from the central fat compartment in the lower eyelids of 10 young and middle-aged patients during routine blepharoplasty surgery.After assessment of the morphological changes of adipocytes with aging,OASCs were isolated from the fat samples and expanded in vitro.Differences in the stem cell colony number(fibroblast colony-forming unit),growth rate and phenotype characterization(flow cytometry analysis)were evaluated.The ability of OASCs to differentiate into adipocytes was determined by oil red O staining and the mRNA expression level of peroxisome proliferator-activated receptorγ.Results:Fat cell size showed a decreasing trend with advancing age.Although no difference was found in the expression of cell surface markers,the colony number and proliferative rate of OASCs from middle-aged donors were significantly lower than those from the young donors.The adipogenic differentiation capacity of middle-aged OASCs was also reduced.These differences were statistically significant(P<0.001).Conclusion:The data showed that the progenitor cell number,proliferation capacity and adipogenic potential of OASCs decreased with aging,suggesting that using OASCs from elderly patients for therapeutic purposes might be restricted.
文摘During excessive adipose tissue accumulation,various adipokines such as visfatin,chemerin,vaspin,and adiponectin are released into systemic circulation,thereby influencing metabolic tissue function throughout the body.As multifunctional signaling molecules secreted by adipose tissue,adipokines play a pivotal role in metabolic regulation,inflammatory response,and tissue homeostasis.Recent studies have demonstrated that adipokines can influence skeletal system repair and regeneration by modulating bone marrow-derived mesenchymal stem cell(BMSC)proliferation,differentiation,migration,and immunomodulatory functions.However,different adipokines have distinct roles in regulating BMSC function,but their underlying molecular mechanisms are not fully understood.In this review,we systematically summarize the specific mechanisms of action and potential clinical applications of visfatin,chemerin,vaspin,and adiponectin on BMSC function in order to reveal new mechanisms of interaction between adipokines and BMSCs.The aim is to provide a theoretical basis for targeted treatment strategies for bone diseases targeting adipokines.
基金supported by the National key research and development program of China(Grant No.2018YFD0500402)the National Natural Science Foundation of China(Grant No.31790412,Grant No.31672431)the National Key Basic Research Program of China(2013CB127302。
文摘Background:Long non-coding RNAs(lncRNAs)are emerging key regulators involved in a variety of biological processes such as cell differentiation and development.The balance between myogenesis and adipogenesis is crucial for skeletal muscle homeostasis in humans and meat quality in farm animals.The present study aimed to reveal the global transcriptomic profiles of adipogenic(Adi-)and myogenic(Myo-)precursors derived from porcine skeletal muscle and identify lncRNAs involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Results:In this study,a total of 655 novel individual lncRNAs including differentially expressed 24 lncRNAs,and 755 differentially expressed mRNAs were identified(fold change≥2 or≤0.5 and adjusted P<0.05).Integrated results of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis accompanied by the variation of intracellular Ca2+concentration highlighted Lnc-ADAMTS9 involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Although Lnc-ADAMTS9 knock-down did not alter the mRNA expression of ADAMTS9,we demonstrated that Lnc-ADAMTS9 can promote myogenic proliferation and myogenic differentiation of myogenic precursors through inhibiting the ERK/MAPK signaling pathway.Conclusion:We deciphered a comprehensive catalog of mRNAs and lncRNAs that might be involved in the regulation of myogenesis and adipogenesis homeostasis in the skeletal muscle of pigs.The Lnc-ADAMTS9 exerts an essential role in myogenesis through the ERK signaling pathway.
