Objective: In traditional Chinese medicine, the herbal pair, Radix Achyranthis Bidentatae(RAB) and Eucommiae Cortex(EC), is widely used to treat osteoporosis. Herein, we determined whether this herbal pair can be used...Objective: In traditional Chinese medicine, the herbal pair, Radix Achyranthis Bidentatae(RAB) and Eucommiae Cortex(EC), is widely used to treat osteoporosis. Herein, we determined whether this herbal pair can be used to ameliorate glucocorticoid(GC)-induced osteoporosis(GIOP) and find its optimal dosage in zebrafish.Methods: The characteristics of the aqueous extract of RAB and EC were separately characterized using high-performance liquid chromatography. Osteoporosis was induced in 5-day post-fertilization zebrafish larvae by exposing them to 10 lmol/L dexamethasone(Dex) for 96 h. Seven combinations of different ratios of RAB and EC were co-administered. Treatment efficacy was determined by calculating zebrafish vertebral area and sum brightness, via alizarin red staining, and by detecting alkaline phosphatase(ALP)activity. Multiple regression analysis was conducted to test the optimal dosage ratio.Results: According to the Chinese Pharmacopoeia(2015), b-ecdysone(b-Ecd) is a major bioactive marker in RAB extract, while pinoresinol diglucoside(PDG) is the major marker in EC extract. Both of b-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards. Treatment with 10 lmol/L Dex reduced zebrafish vertebral area, sum brightness, and ALP activity, but RAB and EC attenuated these effects. Combining 50 mg/m L RAB and 50 mg/m L EC was optimal for preventing GIOP in zebrafish.Reverse transcription-quantitative polymerase chain reaction was used to evaluate the m RNA expression of osteogenesis-related genes. A treatment of 10 lmol/L Dex decreased runt-related transcription factor 2(Runx2), osteogenic protein-1(OP-1), bone c-carboxyglutamic acid-containing protein(BGLAP), and bcatenin levels. This effect was counteracted by RAB and EC co-treatment(P < 0.05). Additionally, the effect of using the two herbal extracts together was better than single-herb treatments separately. These results demonstrated that RAB and EC preserve osteoblast function in the presence of GC. The best mass ratio was 1:1.Conclusion: RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish, with 1:1 as the optimal dosage ratio.展开更多
目的支气管哮喘是严重影响全球公众健康的重大慢病之一,通过使用GEO数据集和孟德尔随机化方法确定哮喘新的遗传靶点,为临床治疗和机制研究提供依据。方法通过基因表达综合数据库(GEO)获得相关数据集,获得数据后进行差异基因的表达数量...目的支气管哮喘是严重影响全球公众健康的重大慢病之一,通过使用GEO数据集和孟德尔随机化方法确定哮喘新的遗传靶点,为临床治疗和机制研究提供依据。方法通过基因表达综合数据库(GEO)获得相关数据集,获得数据后进行差异基因的表达数量性状位点(expression quantitative trait locus,eQTL)分析和孟德尔随机化(mendelian randomization,MR)分析,确定潜在靶点;再通过基因集合富集分析(gene set enrichment analysis,GSEA)和基因本体论(gene ontology,GO)/京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析来探索这些基因的功能和富集通路;通过免疫浸润方法探索靶点与相关免疫细胞的关联;利用医学本体信息检索平台Coremine Medical数据库,筛选核心基因相关治疗中药并进行归纳分析,最后设立外部验证集进行验证。结果共鉴定出280个高表达基因和1127个低表达基因;MR分析确定了12个与哮喘显著相关的核心基因靶点:PGAP3、FAM177A1、UGDH、AASDH、CREB1、ZNF429、CCNG2、SKAP2、ANKRD10、DR1、ISOC1以及LPAR6;预测出人参、五味子、麻黄、杜仲、北沙参等67味靶向中药,主要涉及补虚药、活血化瘀药;MR分析结果与外部验证集的结果一致,强调了本研究的可靠性。结论筛选并验证了12个哮喘潜在靶点,并对相关干预中药进行了预测,为进一步深入探究哮喘的发病机制、早期筛查诊断、早期预防、靶向治疗以及中医药临床诊疗提供了新的线索。展开更多
基金funded by the Natural Science Foundation of China(No.81873318)the Program Foundation of Shanghai Health and Family Planning Commission,China(No.2018JP013)+1 种基金the Ph.D.Program Foundation of Shanghai Municipal Education Commission,China(SMEC,No.B201713)the Postgraduate Innovation Program of Shanghai University of TCM(2020JX357)。
文摘Objective: In traditional Chinese medicine, the herbal pair, Radix Achyranthis Bidentatae(RAB) and Eucommiae Cortex(EC), is widely used to treat osteoporosis. Herein, we determined whether this herbal pair can be used to ameliorate glucocorticoid(GC)-induced osteoporosis(GIOP) and find its optimal dosage in zebrafish.Methods: The characteristics of the aqueous extract of RAB and EC were separately characterized using high-performance liquid chromatography. Osteoporosis was induced in 5-day post-fertilization zebrafish larvae by exposing them to 10 lmol/L dexamethasone(Dex) for 96 h. Seven combinations of different ratios of RAB and EC were co-administered. Treatment efficacy was determined by calculating zebrafish vertebral area and sum brightness, via alizarin red staining, and by detecting alkaline phosphatase(ALP)activity. Multiple regression analysis was conducted to test the optimal dosage ratio.Results: According to the Chinese Pharmacopoeia(2015), b-ecdysone(b-Ecd) is a major bioactive marker in RAB extract, while pinoresinol diglucoside(PDG) is the major marker in EC extract. Both of b-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards. Treatment with 10 lmol/L Dex reduced zebrafish vertebral area, sum brightness, and ALP activity, but RAB and EC attenuated these effects. Combining 50 mg/m L RAB and 50 mg/m L EC was optimal for preventing GIOP in zebrafish.Reverse transcription-quantitative polymerase chain reaction was used to evaluate the m RNA expression of osteogenesis-related genes. A treatment of 10 lmol/L Dex decreased runt-related transcription factor 2(Runx2), osteogenic protein-1(OP-1), bone c-carboxyglutamic acid-containing protein(BGLAP), and bcatenin levels. This effect was counteracted by RAB and EC co-treatment(P < 0.05). Additionally, the effect of using the two herbal extracts together was better than single-herb treatments separately. These results demonstrated that RAB and EC preserve osteoblast function in the presence of GC. The best mass ratio was 1:1.Conclusion: RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish, with 1:1 as the optimal dosage ratio.
文摘目的支气管哮喘是严重影响全球公众健康的重大慢病之一,通过使用GEO数据集和孟德尔随机化方法确定哮喘新的遗传靶点,为临床治疗和机制研究提供依据。方法通过基因表达综合数据库(GEO)获得相关数据集,获得数据后进行差异基因的表达数量性状位点(expression quantitative trait locus,eQTL)分析和孟德尔随机化(mendelian randomization,MR)分析,确定潜在靶点;再通过基因集合富集分析(gene set enrichment analysis,GSEA)和基因本体论(gene ontology,GO)/京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析来探索这些基因的功能和富集通路;通过免疫浸润方法探索靶点与相关免疫细胞的关联;利用医学本体信息检索平台Coremine Medical数据库,筛选核心基因相关治疗中药并进行归纳分析,最后设立外部验证集进行验证。结果共鉴定出280个高表达基因和1127个低表达基因;MR分析确定了12个与哮喘显著相关的核心基因靶点:PGAP3、FAM177A1、UGDH、AASDH、CREB1、ZNF429、CCNG2、SKAP2、ANKRD10、DR1、ISOC1以及LPAR6;预测出人参、五味子、麻黄、杜仲、北沙参等67味靶向中药,主要涉及补虚药、活血化瘀药;MR分析结果与外部验证集的结果一致,强调了本研究的可靠性。结论筛选并验证了12个哮喘潜在靶点,并对相关干预中药进行了预测,为进一步深入探究哮喘的发病机制、早期筛查诊断、早期预防、靶向治疗以及中医药临床诊疗提供了新的线索。
