Objective To investigate the Wnt signaling pathway and miRNAs mechanism of extracts of Plastrum Testudinis(PT)in the treatment of osteoporosis(OP).Methods Thirty female Sprague Dawley rats were randomly divided into 5...Objective To investigate the Wnt signaling pathway and miRNAs mechanism of extracts of Plastrum Testudinis(PT)in the treatment of osteoporosis(OP).Methods Thirty female Sprague Dawley rats were randomly divided into 5 groups by random number table method,including sham group,ovariectomized group(OVX),ovariectomized groups treated with high-,medium-,and low-dose PT(160,80,40 mg/kg per day,respectively),with 6 rats in each group.Except for the sham group,the other rats underwent bilateral ovariectomy to simulate OP and received PT by oral gavage for 10 consecutive weeks.After treatment,bone mineral density was measured by dual-energy X-ray absorptiometry;bone microstructure was analyzed by micro-computed tomography and hematoxylin and eosin staining;and the expressions of osteogenic differentiation-related factors were detected by immunochemistry,Western blot,and quantitative polymerase chain reaction.In addition,Dickkopf-1(Dkk-1)was used to inhibit the Wnt signaling pathway in bone marrow mesenchymal stem cells(BMSCs)and miRNA overexpression was used to evaluate the effect of miR-214 on the osteogenic differentiation of BMSCs.Subsequently,PT extract was used to rescue the effects of Dkk-1 and miR-214,and its impacts on the osteogenic differentiation-related factors of BMSCs were evaluated.展开更多
OBJECTIVE:To investigate the effect of Guijiajiao(Colla Carapacis et Plastri, CCP), the glue of tortoise shell, on bone regeneration in zebrafish and male Wistar rats.METHODS: In this study, we applied in vitro and in...OBJECTIVE:To investigate the effect of Guijiajiao(Colla Carapacis et Plastri, CCP), the glue of tortoise shell, on bone regeneration in zebrafish and male Wistar rats.METHODS: In this study, we applied in vitro and in vivo models, tissue section analysis, and quantitative polymerase chain reaction to assess the effects of CCP on bone repair. MG-63 cells were used in alkaline phosphatase(ALP) activity and mineralization assays. Zebrafish and male Wistar rats were used to evaluate the effects of CCP on bone repair in vivo.RESULTS: A simple preparation of CCP promoted osteogenesis in vivo and in vitro, and promoted MG-63 cell proliferation as well as ALP activity and mineralization. In addition, CCP activated Akt and extracellular signaling-regulated kinase pathways and significantly increased the expression of ossification-related genes and proteins such as runt-related transcription factor-2, osteocalcin, and osteopontin. The in vivo results revealed promotion of osteogenesis by immersing zebrafish in CCP for 72 h.An oral dose of 1.25 g/kg CCP significantly improved skull defects in rats, which was accompanied by an increase in serum ALP levels.CONCLUSION: One of the ingredients of Guilu Erxianjiao(tortoise shell and deer horn formula) pro-vides a practical alternative therapy for bone regeneration.展开更多
基金Supported by the National Natural Science Foundation of China(No.U24A6013,No.82274232 and No.82405121)Science and Technology Program Project of Guangdong Province-Guangdong Provincial Key Laboratory of Traditional Chinese Informatization(No.2021B1212040007)+2 种基金Basic and Applied Basic Research Fund of Guangdong Province(No.2022B1515120022)Construction Project of Guangdong Famous Traditional Chinese Medicine Inheritance Studio of ZHANG Rong-hua[Guangdong Traditional Chinese Medicine Letter(2023)No.108]China Association for Science and Technology(CAST)Young Talent Support Project Doctoral Student Special Program(CAST Office Letter[2024]No.92)。
文摘Objective To investigate the Wnt signaling pathway and miRNAs mechanism of extracts of Plastrum Testudinis(PT)in the treatment of osteoporosis(OP).Methods Thirty female Sprague Dawley rats were randomly divided into 5 groups by random number table method,including sham group,ovariectomized group(OVX),ovariectomized groups treated with high-,medium-,and low-dose PT(160,80,40 mg/kg per day,respectively),with 6 rats in each group.Except for the sham group,the other rats underwent bilateral ovariectomy to simulate OP and received PT by oral gavage for 10 consecutive weeks.After treatment,bone mineral density was measured by dual-energy X-ray absorptiometry;bone microstructure was analyzed by micro-computed tomography and hematoxylin and eosin staining;and the expressions of osteogenic differentiation-related factors were detected by immunochemistry,Western blot,and quantitative polymerase chain reaction.In addition,Dickkopf-1(Dkk-1)was used to inhibit the Wnt signaling pathway in bone marrow mesenchymal stem cells(BMSCs)and miRNA overexpression was used to evaluate the effect of miR-214 on the osteogenic differentiation of BMSCs.Subsequently,PT extract was used to rescue the effects of Dkk-1 and miR-214,and its impacts on the osteogenic differentiation-related factors of BMSCs were evaluated.
文摘OBJECTIVE:To investigate the effect of Guijiajiao(Colla Carapacis et Plastri, CCP), the glue of tortoise shell, on bone regeneration in zebrafish and male Wistar rats.METHODS: In this study, we applied in vitro and in vivo models, tissue section analysis, and quantitative polymerase chain reaction to assess the effects of CCP on bone repair. MG-63 cells were used in alkaline phosphatase(ALP) activity and mineralization assays. Zebrafish and male Wistar rats were used to evaluate the effects of CCP on bone repair in vivo.RESULTS: A simple preparation of CCP promoted osteogenesis in vivo and in vitro, and promoted MG-63 cell proliferation as well as ALP activity and mineralization. In addition, CCP activated Akt and extracellular signaling-regulated kinase pathways and significantly increased the expression of ossification-related genes and proteins such as runt-related transcription factor-2, osteocalcin, and osteopontin. The in vivo results revealed promotion of osteogenesis by immersing zebrafish in CCP for 72 h.An oral dose of 1.25 g/kg CCP significantly improved skull defects in rats, which was accompanied by an increase in serum ALP levels.CONCLUSION: One of the ingredients of Guilu Erxianjiao(tortoise shell and deer horn formula) pro-vides a practical alternative therapy for bone regeneration.
