This study aims to investigate the pharmacological effects and mechanisms of Yougui Yin in treating glucocorticoid-induced osteonecrosis. A rat model of glucocorticoid-associated osteonecrosis of the femoral head(GA-ONFH) was established by intramuscular injection of dexamethasone at 20 mg·kg~(-1) every other day for 8 weeks. Rats were randomly allocated into control, model, and low-and high-dose(1.5 and 3.0 g·kg~(-1), respectively) Yougui Yin groups. After modeling, rats in Yougui Yin groups were administrated with Yougui Yin via gavage, which was followed by femoral specimen collection. Hematoxylin-eosin staining was employed to observe femoral head repair, and immunofluorescence was employed to assess adipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs) within the femoral head. Cell experiments were carried out with dexamethasone(1 μmol·L~(-1))-treated BMSCs to evaluate the effects of Yougui Yin-medicated serum on adipogenic differentiation. Animal experiments demonstrated that compared with the model group, Yougui Yin at both high and low doses significantly improved bone mineral density(BMD), bone volume/total volume(BV/TV) ratio, and trabecular thickness(Tb.Th) in the femoral head. Additionally, Yougui Yin alleviated necrosis-like changes and adipocyte infiltration and significantly reduced the expression level of peroxisome proliferator-activated receptor γ(PPARγ) in the femoral head, thereby suppressing the adipogenic differentiation of BMSCs in GA-ONFH rats. The cell experiments revealed that Yougui Yin-medicated serum markedly inhibited dexamethasone-induced adipogenic differentiation of BMSCs and down-regulated the level of PPARγ. The overexpression of PPARγ attenuated the inhibitory effect of Yougui Yin-medicated serum on the adipogenic differentiation of BMSCs, indicating the critical role of PPARγ in Yougui Yin-mediated suppression of adipogenic differentiation of BMSCs. In conclusion, Yougui Yin exerts therapeutic effects on glucocorticoid-induced osteonecrosis by down-regulating PPARγ expression and inhibiting adipogenic differentiation of BMSCs.
Animals ; Mesenchymal Stem Cells/metabolism* ; PPAR gamma/genetics* ; Rats ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Glucocorticoids/adverse effects* ; Rats, Sprague-Dawley ; Adipogenesis/drug effects* ; Osteonecrosis/genetics* ; Cell Differentiation/drug effects* ; Bone Marrow Cells/metabolism* ; Femur Head Necrosis/chemically induced* ; Humans

Saponins associated with Panax notoginseng (P. notoginseng) demonstrate significant therapeutic efficacy across multiple diseases. However, certain high-yield saponins face limited clinical applications due to their reduced pharmacological efficacy. This study synthesized and evaluated 36 saponin-1,2,3-triazole derivatives of ginsenosides Rg1/Rb1 and notoginsenoside R1 for anti-adipogenesis activity in vitro. The research revealed that the ginsenosides Rg1-1,2,3-triazole derivative a17 demonstrates superior adipogenesis inhibitory effects. Structure-activity relationships (SARs) analysis indicates that incorporating an amidyl-substituted 1,2,3-triazole into the saponin side chain via Click reaction enhances anti-adipogenesis activity. Additionally, several other derivatives exhibit general adipogenesis inhibition. Compound a17 demonstrated enhanced potency compared to the parent ginsenoside Rg1. Mechanistic investigations revealed that a17 exhibits dose-dependent inhibition of adipogenesis in vitro, accompanied by decreased expression of preadipocytes. Peroxisome proliferator-activated receptor γ (PPARγ), fatty acid synthase (FAS), and fatty acid binding protein 4 (FABP4) adipogenesis regulators. These findings establish the ginsenoside Rg1-1,2,3-triazole derivative a17 as a promising adipocyte differentiation inhibitor and potential therapeutic agent for obesity and associated metabolic disorders. This research provides a foundation for developing effective therapeutic approaches for various metabolic syndromes.
Adipogenesis/drug effects* ; Triazoles/chemical synthesis* ; Ginsenosides/chemical synthesis* ; Saponins/chemical synthesis* ; Animals ; Mice ; Structure-Activity Relationship ; PPAR gamma/genetics* ; 3T3-L1 Cells ; Adipocytes/metabolism* ; Panax notoginseng/chemistry* ; Drug Design ; Molecular Structure ; Humans ; Cell Differentiation/drug effects* ; Fatty Acid-Binding Proteins/genetics*

PURPOSE: Ascorbic acid has been reported to have an adipogenic effect on 3T3-L1 preadipocytes, while evidence also suggests that ascorbic acid reduces body weight in humans. In this study, we tested the effects of ascorbic acid on adipogenesis and the balance of lipid accumulation in ovariectomized rats, in addition to long-term culture of differentiated 3T3-L1 adipocytes. MATERIALS AND METHODS: Murine 3T3-L1 fibroblasts and ovariectomized rats were treated with ascorbic acid at various time points. In vitro adipogenesis was analyzed by Oil Red O staining, and in vivo body fat was measured by a body composition analyzer using nuclear magnetic resonance. RESULTS: When ascorbic acid was applied during an early time point in 3T3-L1 preadipocyte differentiation and after bilateral ovariectomy (OVX) in rats, adipogenesis and fat mass gain significantly increased, respectively. However, lipid accumulation in well-differentiated 3T3-L1 adipocytes showed a significant reduction when ascorbic acid was applied after differentiation (10 days after induction). Also, oral ascorbic acid administration 4 weeks after OVX in rats significantly reduced both body weight and subcutaneous fat layer. In comparison to the results of ascorbic acid, which is a well-known cofactor for an enzyme of collagen synthesis, and the antioxidant ramalin, a potent antioxidant but not a cofactor, showed only a lipolytic effect in well-differentiated 3T3-L1 adipocytes, not an adipogenic effect. CONCLUSION: Taking these results into account, we concluded that ascorbic acid has both an adipogenic effect as a cofactor of an enzymatic process and a lipolytic effect as an antioxidant.
3T3-L1 Cells ; Adipocytes/drug effects ; Adipocytes/metabolism ; Adipogenesis/drug effects ; Animals ; Antioxidants/pharmacology ; Ascorbic Acid/pharmacology ; Body Composition/drug effects ; Body Weight/drug effects ; Cell Differentiation/drug effects ; Female ; Fibroblasts/drug effects ; Fibroblasts/metabolism ; Lipolysis/drug effects ; Mice ; Ovariectomy ; Rats, Sprague-Dawley

Obesity is associated with a number of metabolic abnormalities such as type 2 diabetes and has become a major health problem worldwide. In the present study, we investigated the effects of Epimedium koreanum Nakai (Herba Epimedii, HE) and its main constituent icariin on the adipocyte differentiation in 3T3-L1 preadipocytes. HE extract and icariin significantly reduced lipid accumulation and suppressed the expressions of PPARγ, C/EBPα, and SREBP-1c in 3T3-L1 adipocytes. They also inhibited fatty acid synthase (FAS), acyl-Co A synthase (ACS1), and perilipin. Moreover, HE extract and icariin markedly increased the phosphorylation of AMPK. These results indicated that HE extract and icariin can inhibit the adipocyte differentiation through downregulation of the adipogenic transcription factors, suggesting that HE containing icariin may be used as a potential therapeutic agent in the treatment and prevention of obesity.
3T3-L1 Cells ; Adipocytes ; cytology ; drug effects ; metabolism ; Adipogenesis ; drug effects ; Animals ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; metabolism ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Lipid Metabolism ; drug effects ; Mice ; PPAR gamma ; genetics ; metabolism ; Plant Extracts ; pharmacology ; Sterol Regulatory Element Binding Protein 1 ; genetics ; metabolism

OBJECTIVETo investigate the inhibitory effects of tetramethoxystilbene, a selective CYP1B1 inhibitor, on adipogenic differentiation of C3H10T1/2 multi-potent mesenchymal cells.

METHODSIn vitro cultured C3H10T1/2 cells at full confluence were induced by adipogenic agents (10 µg/ml insulin, 2 µmol/L dexamethasone and 0.5 mmol/L 3-isobutyl-1-methylxanthine) and exposed simultaneously to TMS at the final concentrations of 1.0, 2.0 or 4.0 µg/ml. Oil Red-O staining was used to observe the cell differentiation. The expression of peroxisome proliferator-activated receptor gamma (PPARγ) and its target genes cluster of differentiation 36 (CD36) and fatty acid binding protein 4 (FABP4) were quantified by real-time RT-PCR and Western blotting.

RESULTSOil Red-O staining and TG contents revealed that TMS suppressed induced differentiation of C3H10T1/2 cells. TMS exposure of the cells dose-dependently decreased both mRNA and protein expressions of PPARγ, a key nuclear transcription factor during adipogenesis, and also lowered the mRNA expressions of PPARγ target genes CD36 and FABP4.

CONCLUSIONTMS can suppress adipogenic differentiation of C3H10T1/2 cells by inhibiting PPARγ

Adipogenesis ; drug effects ; Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Cytochrome P-450 CYP1B1 ; Cytochrome P-450 Enzyme Inhibitors ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Mice, Inbred C3H ; PPAR gamma ; metabolism ; Pluripotent Stem Cells ; cytology ; drug effects ; RNA, Messenger ; Stilbenes ; pharmacology

OBJECTIVETo evaluate the influence of adipose tissue extract on inducing angiogenesis and adipogenesis in adipose tissue engineering chamber in vivo.

METHODS6 months' healthy New Zealand rabbits (n = 64) were picked. The inguinal fat pads were cultured, centrifuged, filtered, and the liquid was called adipose tissue extract (ATE). Two adipose tissue engineering chamber were built in the rabbit's back. A week later, 0.2 ml normal saline (control group, left) and 0. 2 ml ATE (experimental group, right) was respectively injected into the chamber. The contents were evaluated morphometrically, histologically and immunohistochemically 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks and 7 weeks after injection. 8 rabbits were observed each time. The data regarding the number of the volume of fat flap and blood capillary at each time point were analyzed by paired t test.

RESULTSAfter injection, new tissue volume was significantly increased in the experimental group [(5.12 ± 0.22) ml], compared with that in control group [(4.90 ± 0.15) ml]. Early angiogenesis was also increased after ATE injection and the total number of capillaries reached peak 1 week after injection, which was (72.80 ± 9.67) in experimental group and (51.40 ± 6.09) in control group. In the mid-term of experimental period, earlier adipogenesis appeared in experimental group. In the later period, the outer capsule of the new construction was thinner in experimental group which reduced the suppression of the adipogenesis.

CONCLUSIONSATE can promote the angiogenesis and adipogenesis in the chamber, and reduce the capsule contracturing, so as to induce the large volume of adipose tissue regeneration

Adipogenesis ; drug effects ; physiology ; Adipose Tissue ; chemistry ; physiology ; Animals ; Neovascularization, Physiologic ; drug effects ; Rabbits ; Regeneration ; Tissue Engineering ; instrumentation ; Tissue Extracts ; pharmacology

PURPOSE: In epidemiologic and animal studies, a high fat diet (HFD) has been shown to be associated with lower bone mineral density (BMD) and a higher risk of osteoporotic fractures. Meanwhile, consuming a HFD containing diacylglycerol (DAG) instead of triacylglycerol (TAG) is known to offer metabolically beneficial effects of reductions in body weight and abdominal fat. The purpose of this study was to investigate the effects of a HFD containing DAG (HFD-DAG) on bone in mice. MATERIALS AND METHODS: Four-week-old male C57BL/6J mice (n=39) were divided into three weight-matched groups based on diet type: a chow diet group, a HFD containing TAG (HFD-TAG) group, and a HFD-DAG group. After 20 weeks, body composition and bone microstructure were analyzed using dual energy X-ray absorptiometry and micro-computed tomography. Reverse transcription-polymerase chain reaction (PCR) and real-time PCR of bone marrow cells were performed to investigate the expressions of transcription factors for osteogenesis or adipogenesis. RESULTS: The HFD-DAG group exhibited lower body weight, higher BMD, and superior microstructural bone parameters, compared to the HFD-TAG group. The HFD-DAG group showed increased expression of Runx2 and decreased expression of PPARgamma in bone marrow cells, compared to the HFD-TAG group. The HFD-DAG group also had lower levels of plasma glucose, insulin, total cholesterol, and triglyceride than the HFD-TAG group. CONCLUSION: Compared to HFD-TAG, HFD-DAG showed beneficial effects on bone and bone metabolism in C57BL/6J mice.
Absorptiometry, Photon ; Adipogenesis ; Animals ; Body Composition ; Body Weight ; Bone Density/*drug effects ; Bone Marrow Cells/metabolism ; Diet, High-Fat/*adverse effects ; Dietary Fats/*pharmacology ; Diglycerides/administration & dosage/*adverse effects ; Male ; Mice ; Mice, Inbred C57BL ; Osteogenesis/*drug effects ; Real-Time Polymerase Chain Reaction ; Triglycerides ; X-Ray Microtomography

OBJECTIVE: To determine the time course and potential mechanism of fibroblast growth factor-1 (FGF-1) in the regulation of adipogenesis.
 METHODS: We cultured human Simpson-Golabi-Behmel syndrome (SGBS) pre-adipocytes with recombinant FGF-1 and harvested cells at various stages prior to and during differentiation; at cell proliferation (D-3), confluence (D0), early (D3), middle (D7) and mature (D14) stages of differentiation. We determined lipid accumulation in mature adipocytes by morphological observation and quantitative measurement of oil red O staining. We also examined the expression of adipogenic genes and related markers involved in the Wnt/β-catenin pathway using quantitative Real-time PCR and Western blot.
 RESULTS: Compared to control SGBS cells, treatment with FGF-1 increased lipid accumulation; induced a sustained increase in the mRNA for peroxisome proliferater-activated receptor γ (PPARγ), glyceraldehyde-3-phosphate dehydrogenase (G3PDH), adiponectin and glucose transporter type 4 (GLUT4); and promoted a sustained decrease in expression of markers of the Wnt/β-catenin pathway, β-catenin and transcription factor 4 (TCF4).
 CONCLUSION The adipogenic effects of FGF-1 are apparent throughout the whole priming and differentiation period in human SGBS pre-adipocytes. Furthermore, our results suggest that FGF-1 
promotes adipogenesis, at least in part, via a sustained decrease in activity of the Wnt/β-catenin pathway.
Adipocytes ; drug effects ; metabolism ; Adipogenesis ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; metabolism ; Cell Differentiation ; Cells, Cultured ; Fibroblast Growth Factor 1 ; pharmacology ; Humans ; Recombinant Proteins ; pharmacology ; Transcription Factor 4 ; Transcription Factors ; metabolism ; Wnt Signaling Pathway ; beta Catenin ; metabolism

BACKGROUNDSteroids 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 PPARγ expression in BMSCs.

METHODSCells 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.

RESULTSDexamethasone-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 dexamethasone-treated 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.

CONCLUSIONSThe siRNA adenovirus vector targeting PPARγ can efficiently inhibit steroid-induced adipogenic differentiation in rabbit BMSCs and retain their osteogenic differentiation potential.

Adenoviridae ; genetics ; Adipogenesis ; drug effects ; genetics ; Animals ; Cell Differentiation ; drug effects ; genetics ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; PPAR gamma ; genetics ; metabolism ; pharmacology ; RNA, Small Interfering ; Rabbits ; Steroids

OBJECTIVETo explore the new mechanism of propranolol for treatment of hemangioma and the effects of propranolol on proliferation of hemangioma-derived mesenchymal stem cells ( Hem- MSCs).

METHODSWe isolated Hem-MSCs from hemangioma in the proliferating phase by their selective adhesion to plastic culture dishes. Immunofluorescence staining was used to examine the expression of marker antigens in Hem-MSCs. Human umbilical vein endothelial cells(HUVECs) were used as control. Indiuction of multi-lineage differentiation including osteogenesis and adipogeneis was performed with appropriate medium to identify the multi-lineage differentiation potential. MTT cell counting was used to observe the effects of different concentrations of propranolol on proliferation of Hem-MSCs.

RESULTSHem- MSCs were fibroblast-like morphology. All of them expressed vimentin, most expressed α-SMA,CD133, some expressed Glutl, and none of them expressed VEGF. Osteogenic, adipogenic differentiations of Hem- MSCs were induced successfully. Effects of low concentration of propranolol on proliferation of Hem-MSCs were not obvious, while high concentration of propranolol can inhibit the proliferation of Hem-MSCs.

CONCLUSIONSThe cells we isolated from hemangioma are Hem-MSCs. High concentration of propranolol can inhibit the proliferation of Hem-MSCs.

Adipogenesis ; Antigens ; metabolism ; Cell Differentiation ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelium, Vascular ; cytology ; Fibroblasts ; cytology ; Hemangioma ; pathology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; Propranolol ; pharmacology ; Umbilical Veins ; Vimentin ; metabolism