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OBJECTIVES: This study aims to investigate the effects of Traditional Chinese medicine, Periplaneta americana extract (PAE), on osteoblast differentiation of human alveolar bone marrow-derived mesenchymal stem cells (hABMMSCs). MATERIALS AND METHODS: Human alveolar bone marrow-derived mesenchymal stem cells were treated with different concentrations of PAE. Cell Counting Kit-8 (CCK-8) assay and transwell migration assay were conducted to evaluate cell proliferation and migration, respectively. Alkaline phosphatase (ALP) staining, ALP activity assay, and Alizarin red S staining were performed to detect osteogenesis in hABMMSCs. In addition, real-time quantitative polymerase chain reaction (RT-qPCR) and western blot (WB) assay were performed to evaluate expression levels of osteogenic markers. Finally, RNA sequencing analysis and WB were carried out to elucidate the underlying mechanism. RESULTS: A total of 0.1 mg/ml PAE promoted cell proliferation and migration. PAE also increased ALP activity and mineralized nodule formation of hABMMSCs. In addition, PAE upregulated the expression of osteogenesis-related genes (RUNX2, COL1A1, and BGLAP). RNA-sequencing analysis revealed that PAE activated the focal adhesion signaling pathway. Treatment with Defactinib, an inhibitor of FAK, attenuated the effects induced by PAE. CONCLUSIONS: PAE could enhance osteoblast differentiation of hABMMSCs through focal adhesion signaling pathway, suggesting a therapeutic potential for the alveolar bone defect.
Assuntos
Células-Tronco Mesenquimais , Periplaneta , Animais , Humanos , Osteogênese , Diferenciação Celular , Osteoblastos , Células CultivadasRESUMO
HFD (high-fat diet) induces obesity and metabolic disorders, which is associated with the alteration in gut microbiota profiles. However, the underlying molecular mechanisms of the processes are poorly understood. In this study, we used the simple model organism honey bee to explore how different amounts and types of dietary fats affect the host metabolism and the gut microbiota. Excess dietary fat, especially palm oil, elicited higher weight gain, lower survival rates, hyperglycemic, and fat accumulation in honey bees. However, microbiota-free honey bees reared on high-fat diets did not significantly change their phenotypes. Different fatty acid compositions in palm and soybean oil altered the lipid profiles of the honey bee body. Remarkably, dietary fats regulated lipid metabolism and immune-related gene expression at the transcriptional level. Gene set enrichment analysis showed that biological processes, including transcription factors, insulin secretion, and Toll and Imd signaling pathways, were significantly different in the gut of bees on different dietary fats. Moreover, a high-fat diet increased the relative abundance of Gilliamella, while the level of Bartonella was significantly decreased in palm oil groups. This study establishes a novel honey bee model of studying the crosstalk between dietary fat, gut microbiota, and host metabolism.
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Abelhas/fisiologia , Dieta Hiperlipídica , Ácidos Graxos/administração & dosagem , Microbioma Gastrointestinal , Animais , Abelhas/microbiologia , Gorduras na Dieta/administração & dosagem , Regulação da Expressão Gênica , Glucose/química , Insulina/metabolismo , Metabolismo dos Lipídeos , Síndrome Metabólica/metabolismo , Óleo de Palmeira/química , Fenótipo , RNA Ribossômico 16S/metabolismo , Transdução de Sinais , Óleo de Soja/química , Trealose/químicaRESUMO
Chlorogenic acid (CGA), a bioactive compound commonly found in plants, has been demonstrated possessing nutraceutical potential in recent years. However, the more critical issue concerning how to improve production efficacy of CGA is still limited. It is a challenge to harvest a large amount of CGA without prolonging extraction time. In this study, the feasibility of using ultrasound for CGA extraction from Lonicera japonica was investigated. A central composite design (CCD) was employed to evaluate the effects of the operation parameters, including temperature, ethanol concentration, liquid to solid ratio, and ultrasound power on CGA yields. Meanwhile, the process of ultrasound-assisted extraction was optimized through modeling response surface methodology (RSM) and artificial neural network (ANN). The data indicated that CGA was efficiently extracted from the flower of Lonicera japonica by ultrasound assistance. The optimal conditions for the maximum extraction of CGA were as follows: The temperature at 33.56 °C, ethanol concentration at 65.88%, L/S ratio at 46:1 mL/g and ultrasound power at 150 W. ANN possessed greater optimization capacity than RSM for fitting experimental data and predicting the extraction process to obtain a maximum CGA yield. In conclusion, the process of ultrasound-assisted extraction can be well established by a methodological approach using either RSM or ANN, but it is worth mentioning that the ANN model used here showed the superiority over RSM for predicting and optimizing.
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Ácido Clorogênico/isolamento & purificação , Lonicera/química , Extratos Vegetais/química , Fracionamento Químico/métodos , Ácido Clorogênico/química , Etanol/química , Flores/química , Redes Neurais de Computação , UltrassomRESUMO
@#Bone morphogenetic protein-2 (BMP-2) and basic fibroblast growth factor (bFGF) are two important signaling molecules for natural bone repair that are widely applied in the field of bone tissue engineering. BMP-2 accelerates the maturation and mineralization of osteoblasts, and bFGF exhibits significant effects on the promotion of cell division and angiogenesis. The osteogenesis mechanism of BMP-2 differs from that of bFGF, leading to a potentially complementary role between the two proteins. The use of a suitable dose of bFGF in combination with BMP-2 in bone tissue engineering synergistically promotes the formation of new bone and exhibits a better repair effect than either single growth factor. However, the suitable dose range of BMP-2 and bFGF in combination should be further clarified. The mechanism of the synergism and antagonism between BMP-2 and bFGF must be further delineated, and a drug delivery system that better simulates the growth factor release pattern during natural bone repair remains to be designed.
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Several novel oxazolidinone antibiotics with a spiropiperazinyl substituent at the 4'-position of the phenyl ring were synthesized through nitroso Diels-Alder chemistry and the in vitro antibacterial activities were evaluated against various Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis), Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) and mycobacteria (Mycobacterium vaccae, Mycobacterium tuberculosis). Analogs (8a and 12) were active against selected drug resistant microbes, like methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) and had no mammalian toxicity in a Hep-2 cellular assay (CC(50) >100 µM).