RESUMEN
Phytoestrogens are a class of potential natural medicines for treating postmenopausal osteoporosis (PMOP). Segetalin B (SB) is a cyclic peptide compound showing estrogenic activity. This study reports the effect of SB on bone formation among ovariectomized (OVX) rats. The bone marrow mesenchymal stem cells (BMSCs) from OVX rats were cultured in vitro. Alizarin Red staining was utilized to observe the effect of SB on the mineralization of BMSCs. The levels of alkaline phosphatase (ALP), osteocalcin, bone morphogenetic protein (BMP-2), and Sirtuin 1 (SIRT1) activities were detected. The OVX rats were treated with SB in vivo. Micro-CT was utilized for imaging analysis. Urine calcium and phosphorus, and ALP activity in bone marrow were assayed. Western blot analysis and immunofluorescence were incorporated to detect protein expressions in vitro and in vivo. The results showed that SB dose-dependently promoted mineralization of OVX rat-derived BMSCs in vitro increased the level of Osteocalcin, BMP-2, ALP, and SIRT1 activity. Moreover, it upregulated expressions of Runx2, Osterix, and SIRT1, downregulated expressions of Notch intracellular domain (NICD), acetyl-NICD, and hairy and enhancer of split 1 (Hes1). In addition, SB treatment significantly decreased bone loss, inhibited calcium and phosphorus loss, elevated ALP activity, upregulated Runx2, Osterix, and SIRT1, and downregulated NICD and Hes1 in OVX rats in vivo. However, EX527, a SIRT1-selective inhibitor, could reverse the above effects of SB in vitro or in vivo. These results indicate that SB is a potential natural medicine to improve PMOP. Thus, its mechanism of promoting bone formation involves the SIRT1/Notch1 signaling axis.
Asunto(s)
Osteogénesis , Péptidos Cíclicos , Receptor Notch1 , Sirtuina 1 , Animales , Ratas , Calcio/metabolismo , Diferenciación Celular , Células Cultivadas , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Osteocalcina/metabolismo , Péptidos Cíclicos/farmacología , Fósforo/metabolismo , Receptor Notch1/metabolismo , Sirtuina 1/metabolismoRESUMEN
The hot water extract of Rabdosia rubescens was traditionally used as an antithrombotic medicine. To explore its antithrombotic utility and mechanism, we carried out a series of in vitro and in vivo assays in this study. In vitro platelet aggregation assay showed that the half maximal inhibitory concentration values of aqueous extract of R. rubescens leaves (AERL) inhibiting platelet aggregation induced by thrombin, arachidonic acid, adenosine diphosphate, and platelet-activating factor ranged from 0.12 mg/mL to 1.43 mg/mL. The minimal effective oral dose of AERL inhibiting the rats from forming thrombus was 25 mg/kg. Both in vitro and in vivo actions were correlated with AERL concentration-dependently inhibiting sP-selectin release. In water, AERL formed nanoparticles, and their size depended on the concentration. Docking the five nucleotides, 21 phenolic acids, and four diterpenoids identified by high-performance liquid chromatography-photodiode array detector/(-)electrospray ionization-tandem mass spectrometry analysis into the active site of P-selectin, rosmarinic acid was predicted to be the antithrombotic ingredient of AERL. In flow cytometry analysis, 1 µM of rosmarinic acid effectively inhibited sP-selectin release in arachidonic acid-activated platelets. In a rat model, 5 mg/kg of oral rosmarinic acid effectively inhibited thrombosis.
Asunto(s)
Isodon/química , Selectina-P/metabolismo , Hojas de la Planta/química , Agregación Plaquetaria/efectos de los fármacos , Trombosis/tratamiento farmacológico , Adenosina Difosfato/química , Animales , Ácido Araquidónico/química , Coagulación Sanguínea/efectos de los fármacos , Plaquetas/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Cinamatos/química , Depsidos/química , Citometría de Flujo , Hidroxibenzoatos/química , Espectroscopía de Resonancia Magnética , Masculino , Nanopartículas/química , Extractos Vegetales/química , Factor de Activación Plaquetaria/metabolismo , Inhibidores de Agregación Plaquetaria/química , Ratas , Ratas Wistar , Espectrometría de Masa por Ionización de Electrospray , Trombina/metabolismo , Ácido RosmarínicoRESUMEN
OBJECTIVE: To research the effects of Alligator Zhikegao on relieving cough, dispelling phlegm and anti-inflammation. METHOD: The coughing tests in mice, the phenol red secreting tests in mice, ear edema tests in mice,and paw edema tests and subcutaneous cotton ball granuloma in rats were adopted for observing the related pharmacological effects of Alligator Zhikegao. RESULT: Alligator Zhikegao could obviously prolong the latent period and decrease the times of mouse coughing, and remarkably inhibit the mouse ear edema (P < 0.001), the rat paw edema and the hyperplasia of subcutaneous cotton ball granuloma in rats. Alligator Zhikegao 11.70 g x kg(-1) could significant improve the carbonic clearances of macrophages (P <0.05) and the hemolysin level in serum (P <0.01). CONCLUSION: Alligator Zhikegao has significant effects on relieving cough, dispelling phlegm, anti-inflammation and immunological regulation.