ABSTRACT
Osteoarthritis is a prevalent global joint disease, which is characterized by inflammatory reaction and cartilage degradation. Cyasterone, a sterone derived from the roots of Cyathula officinalis Kuan, exerts protective effect against several inflammation-related diseases. However, its effect on osteoarthritis remains unclear. The current study was designed to investigate the potential anti-osteoarthritis activity of cyasterone. Primary chondrocytes isolated from rats induced by interleukin (IL)-1β and a rat model stimulated by monosodium iodoacetate (MIA) were used for in vitro and in vivo experiments, respectively. The results of in vitro experiments showed that cyasterone apparently counteracted chondrocyte apoptosis, increased the expression of collagen II and aggrecan, and restrained the production of the inflammatory factors inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5), metalloproteinase-3 (MMP-3), and metalloproteinase-13 (MMP-13) induced by IL-1β in chondrocytes. Furthermore, cyasterone ameliorated the inflammation and degenerative progression of osteoarthritis potentially by regulating the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. For in vivo experiments, cyasterone significantly alleviated the inflammatory response and cartilage destruction of rats induced by monosodium iodoacetate, where dexamethasone was used as the positive control. Overall, this study laid a theoretical foundation for developing cyasterone as an effective agent for the alleviation of osteoarthritis.
Subject(s)
Animals , Rats , Chondrocytes , NF-kappa B , Iodoacetic Acid , Inflammation , MAP Kinase Signaling System , ApoptosisABSTRACT
Objective The extraction process of Shenxi Oral Liquid by exploring the optional extraction process parameters based on determining the index weight. Methods Using the contents of cyasterone, ginsenosides Rg1, Re, Rb1, panaxadiol, ratio of dry extraction, total polysaccharides, and total peak area as comprehensive evaluation indexes, the weighting coefficient in eight synthetic evaluation indexes was determined by analytic hierarchy process (AHP), criteria importance through intercriteria correlation (CRITIC) method, and mixed weighted AHP-CRITIC. Compared mixed weighting method and single weighting method, the optimal parameters of compound extraction process were optimized by orthogonal test. Results Mixed weighted AHP-CRITIC was more scientific, reasonable, and comprehensive than the single weighting method. According to the weighting coefficient determined by comprehensive evaluation, the optimal process parameters were as follow: compound herbs plus nine times of water, extracting three times, each for 80 min. The mean of three batches of comprehensive evaluation was 99.06 and RSD value was 0.18%. Conclusion AHP combined with CRITIC could be used to establish weighting coefficient of the compound formula extraction process and the optimized process of extraction had been verified to be reasonable, stable, and reproducible, which could be used for industrial production.
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Objective To explore the influence of cyasterone on the osteoclast and osteoblast differentiation and then to investigate its effect on the bone quality in the osteoporosis mice. Methods CCK8 assay was firstly used to detect the toxic effect of cyasterone on the mouse bone marrow derived mononuclear macrophages (BMMs) and anterior osteoblast lines MC3T3E1. Cell apoptosis was measured by flow cytometry. Then TRAP staining and ALP staining were employed to detect osteoclast differentiation and osteoblast differentiation, respectively. Realtime PCR was carried out to test the expression of osteoclast special gene TRAP and osteogenesis crucial gene ALP. In vivo, 15 mice were divided into three groups: sham-operated group, OVX group and OVX+cyasterone treatment group. In treatment group, cyasterone was used as 5mg/kg every day. Sham-operated group and OVX group were treat with saline solution. After 4 weeks, the tibia was collected for Micro-CT detection to observe the bone quality and microstructure changes. Results Cyasterone with the concentration of less than 10 mg/L had no significant cytotoxicity nor influence on the apoptosis (P＞0.05) . Cyasterone could significantly inhibit the osteoclast differentiation of BMMs (P＜0.05), simultaneously, it also had the effect to promote the osteoblast differetiation of MC3T3E1. Real-time PCR indicated that cyasterone could block the expression of TRAP and increase the expression of ALP (P＜0.05) . In vivo, cyasterone was able to obviously improve the osteoporosis status caused by estrogen deficiency without general toxicity. Conclusion cyasterone could provide a good treatment for osteoporosis through the bidirectional effect of inhibiting osteoclast differetiation and promoting osteoblast differentiation.
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Objective To investigate the chemical constituents of Ajuga ovalifolia var. calantha. Methods The 95% ethanol extract from the whole herb of A. ovalifolia var. calantha was separated and purified by column chromatography over silica gel, Sephadex LH-20, ODS, and RP-HPLC. Their chemical structures were identified on the basis of physicochemical characteristics and spectral analyses. Results Thirteen compounds were isolated and identified as (16S)-12,16-epoxy-11,14-dihydroxy-17 (15→16)-abeo-abieta- 8,11,13-trien-7-one (1), ajuforrestin B (2), ajudecumin A (3), 14,15-dihydroajugapitin (4), cyasterone (5), β-sitosterol (6), acacetin (7), apigenin (8), luteolin (9), scopoletin (10), isoscopoletin (11), 4-hydroxy-3-methoxy-benzaldehyde (12), and acetovanillon (13). Conclusion Compounds 1 and 10-13 are reported from the genus Ajuga Linn., and compounds 2-5 and 7 are isolated from this plant for the first time. Single crystal X-ray diffraction experiment is carried out for compound 1, and the crystal structure data of single crystal X-ray diffraction of compound 1 is obtained for the first time.
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Objective: To reveal the variation regularity of chemical components in crude Cyathula officinalis and its processed products (stir-fried with yellow rice wine and salt), and promote the quality control of the herb. Methods: Agilent Zorbax Eclipse XDB-C18 column (250 mm×4.6 mm, 5 μm) was used with acetonitrile and 0.1% phosphoric acid solution in gradient elution mode. The detective wavelength was 266 nm and the flow rate was 0.5 mL/min. The fingerprints for 24 herbal samples were set up and 25 peaks were recorded with different retention time and peak areas. Three peaks were successfully identified as puerarin, cyasterone, and daidzein by comparing with the retention time of reference substances. The vectorial angle method was used to evaluate the similarity between fingerprints. The cluster analysis and principal component analysis were applied to studying the HPLC fingerprint and chemical pattern recognition. Results: The two processing techniques, stir-frying with wine and salt, both had significant effect on herbal chemical profiles. The contents of three known active components, puerarin, cyasterone, and daidzein, were not observed except a little increasing of the content of puerarin after the crude herbs were processed with salt. Conclusion: More precise active components of the processed products of C. officinalis need to future study to improve the current quality standard of C. officinalis in Chinese Pharmacopoeia. The method provided by this study is a powerful tool to identify and quality control between crude and processed C. officinalis because of its quantificational and visual evaluation system.
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Objective: To establish a method for the determination of cyasterone in Shenshixiao capsules. Methods: Using an HPLC method, the column was an Agilent C18 (250 mm × 4. 6 mm,5 μm) column, the mobile phase was methanol-water with gradient elution. The detection wavelength was 243nm, the flow rate was 1. 0 ml·min-1 ,and the temperature of column was room temperature. Results:The linear relationship of cyasterone was good within the concentration range of 0. 025-0. 247 μg(r=0. 999 6), the average recovery was 99. 4%, and RSD was 0. 76%. Conclusion:The method is rapid, simple and accurate, and can be used in the quality control of the preparation.