目的:优化山茶花瓣、叶和花蕊三部位的黄酮提取量,比较抗氧化活性并分析其化学成分。方法:以总黄酮提取量为评价指标,通过单因素实验和正交试验优化超声波辅助酶解法黄酮提取工艺。采用DPPH、ABTS+自由基清除实验和还原力实验评价优化...目的:优化山茶花瓣、叶和花蕊三部位的黄酮提取量,比较抗氧化活性并分析其化学成分。方法:以总黄酮提取量为评价指标,通过单因素实验和正交试验优化超声波辅助酶解法黄酮提取工艺。采用DPPH、ABTS+自由基清除实验和还原力实验评价优化后总黄酮提取物的抗氧化能力。通过超高效液相色谱-四极杆飞行时间质谱联用技术(UPLC-QTOF-MS/MS)分析各部位的化学成分。结果:在最优提取工艺下,山茶各部位总黄酮提取量依次为叶(63.14 RE mg/g)>花蕊(58.77 RE mg/g)>花瓣(20.26 RE mg/g)。抗氧化活性测定结果表明,花瓣、叶、花蕊中黄酮清除DPPH自由基的IC_(50)值分别为7.28、2.51、1.15 mg/mL,清除ABTS+自由基的IC_(50)值分别为2.85、0.95、0.59 mg/mL,还原力分别为63.25、214.11、475.90 Trolox mg/g,其抗氧化强弱顺序为:花蕊>叶>花瓣。通过UPLC-QTOF-MS/MS从花瓣、叶、花蕊三部位共鉴定出29种化合物,且B型原花青素二聚体、表儿茶素、槲皮素-3-O-半乳糖苷等黄酮单体化合物在三个部位中的分布规律与总黄酮提取量一致。结论:超声波辅助酶法能够有效提高黄酮类化合物的提取量,且山茶的叶和花蕊部位具有较高黄酮含量及抗氧化活性。展开更多
目的阐释古代经典名方“保元汤”的药效物质基础并探讨其分子水平的作用机制,发掘保元汤更多潜在的临床应用优势。方法采用UPLC-QTOF-MS/MS技术,选用Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.7μm)色谱柱,以0.1%甲酸水(A)-乙腈...目的阐释古代经典名方“保元汤”的药效物质基础并探讨其分子水平的作用机制,发掘保元汤更多潜在的临床应用优势。方法采用UPLC-QTOF-MS/MS技术,选用Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.7μm)色谱柱,以0.1%甲酸水(A)-乙腈(B)为流动相系统进行梯度洗脱,高分辨飞行时间质谱电喷雾电离源(ESI),正、负离子扫描模式,检测鉴定保元汤的化学成分组成,并分析保元汤大鼠多次多天ig给药后含药血浆中的原型化学成分及代谢产物,进一步运用网络药理学方法,利用TCMSP数据库和药物信息数据库等分析保元汤入血原型成分的靶向作用,并揭示其治疗潜能。结果根据精确相对分子质量数据和多级质谱碎片离子,结合对照品、数据库以及文献报道,共鉴定出保元汤中133个化学成分,并且确定大鼠ig保元汤后,35个成分能够经口吸收入血达到血浆稳态的药物浓度。保元汤入血原型成分“成分-靶点”网络图、核心靶基因基因本体(gene ontology,GO)功能富集分析、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析、疾病预测分析表明,保元汤可通过人参皂苷Rb1、丹皮酚、刺芒柄花素、刺甘草查耳酮、大豆异黄酮、甘草查耳酮A、6-姜辣素、水杨酸等8个关键成分作用于丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、肿瘤蛋白p53(tumor protein p53,TP53)、信号传导及转录激活蛋白(signal transducer and activator of transcription)等核心靶点,通过糖基化终末产物(advanced glycation end products,AGE)-晚期糖基化终末产物受体(receptor for advanced glycation end-products,RAGE)、表皮生长因子受体(epidermal growth factor receptor,EGFR)、非受体酪氨酸激酶(janus kinase,JAK)-信号转导子和转录激活子(signal transducer and activator of transcription,STAT)等信号通路等途径发挥药效作用,提示保元汤除治疗慢性心力衰竭外,还展开更多
Objective: To identify phytochemical constituents present in the extract of flowers of Xanthoceras sorbifolia and evaluate their anti-oxidant and anti-hyperglycemic capacities.Methods: The AlCl3colorimetric method and...Objective: To identify phytochemical constituents present in the extract of flowers of Xanthoceras sorbifolia and evaluate their anti-oxidant and anti-hyperglycemic capacities.Methods: The AlCl3colorimetric method and Prussian Blue assay were used to determine the contents of total flavonoids and total phenolic acids in extraction layers, and the bioactive layers was screened through anti-oxidative activity in vitro. The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC BEH C18column(2.0 mm × 150 mm, 5 μm) were used to identify the ingredients. And anti-oxidative ingredients were screened by off-line UPLC-QTOF-MS/MS-free radical scavenging. The ameliorative role of it was further evaluated in a high-fat, streptozotocin-induced type 2 diabetic rat model and the study was carried out on NADPH oxidase(PDB ID: 2CDU) by molecular docking.Results: Combined with the results of activity screening in vitro, the anti-oxidative part was identified as the ethyl acetate layer. A total of 24 chemical constituents were identified by liquid chromatographymass spectrometry in the ethyl acetate layer and 13 main anti-oxidative active constituents were preliminarily screened out through off-line UPLC-QTOF-MS/MS-free radical scavenging. In vivo experiments showed that flowers of X. sorbifolia could significantly reduce the blood glucose level of diabetic mice and alleviate liver cell damage. Based on the results of docking analysis related to the identified phytocompounds and oxidase which involved in type 2 diabetes, quercetin 3-O-rutinoside, kaempferol-3-O-rhamnoside, isorhamnetin-3-O-glucoside, and isoquercitrin showed a better inhibitory profile.Conclusion: The ethyl acetate layer was rich in flavonoids and phenolic acids and had significant anti-oxidant activity, which could prevent hyperglycemia. This observed activity profile suggested X. sorbifolia flowers as a promising new source of tea to develop alternative natural anti-diabetic products with a high safety margin.展开更多
文摘目的:优化山茶花瓣、叶和花蕊三部位的黄酮提取量,比较抗氧化活性并分析其化学成分。方法:以总黄酮提取量为评价指标,通过单因素实验和正交试验优化超声波辅助酶解法黄酮提取工艺。采用DPPH、ABTS+自由基清除实验和还原力实验评价优化后总黄酮提取物的抗氧化能力。通过超高效液相色谱-四极杆飞行时间质谱联用技术(UPLC-QTOF-MS/MS)分析各部位的化学成分。结果:在最优提取工艺下,山茶各部位总黄酮提取量依次为叶(63.14 RE mg/g)>花蕊(58.77 RE mg/g)>花瓣(20.26 RE mg/g)。抗氧化活性测定结果表明,花瓣、叶、花蕊中黄酮清除DPPH自由基的IC_(50)值分别为7.28、2.51、1.15 mg/mL,清除ABTS+自由基的IC_(50)值分别为2.85、0.95、0.59 mg/mL,还原力分别为63.25、214.11、475.90 Trolox mg/g,其抗氧化强弱顺序为:花蕊>叶>花瓣。通过UPLC-QTOF-MS/MS从花瓣、叶、花蕊三部位共鉴定出29种化合物,且B型原花青素二聚体、表儿茶素、槲皮素-3-O-半乳糖苷等黄酮单体化合物在三个部位中的分布规律与总黄酮提取量一致。结论:超声波辅助酶法能够有效提高黄酮类化合物的提取量,且山茶的叶和花蕊部位具有较高黄酮含量及抗氧化活性。
文摘目的阐释古代经典名方“保元汤”的药效物质基础并探讨其分子水平的作用机制,发掘保元汤更多潜在的临床应用优势。方法采用UPLC-QTOF-MS/MS技术,选用Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.7μm)色谱柱,以0.1%甲酸水(A)-乙腈(B)为流动相系统进行梯度洗脱,高分辨飞行时间质谱电喷雾电离源(ESI),正、负离子扫描模式,检测鉴定保元汤的化学成分组成,并分析保元汤大鼠多次多天ig给药后含药血浆中的原型化学成分及代谢产物,进一步运用网络药理学方法,利用TCMSP数据库和药物信息数据库等分析保元汤入血原型成分的靶向作用,并揭示其治疗潜能。结果根据精确相对分子质量数据和多级质谱碎片离子,结合对照品、数据库以及文献报道,共鉴定出保元汤中133个化学成分,并且确定大鼠ig保元汤后,35个成分能够经口吸收入血达到血浆稳态的药物浓度。保元汤入血原型成分“成分-靶点”网络图、核心靶基因基因本体(gene ontology,GO)功能富集分析、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析、疾病预测分析表明,保元汤可通过人参皂苷Rb1、丹皮酚、刺芒柄花素、刺甘草查耳酮、大豆异黄酮、甘草查耳酮A、6-姜辣素、水杨酸等8个关键成分作用于丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、肿瘤蛋白p53(tumor protein p53,TP53)、信号传导及转录激活蛋白(signal transducer and activator of transcription)等核心靶点,通过糖基化终末产物(advanced glycation end products,AGE)-晚期糖基化终末产物受体(receptor for advanced glycation end-products,RAGE)、表皮生长因子受体(epidermal growth factor receptor,EGFR)、非受体酪氨酸激酶(janus kinase,JAK)-信号转导子和转录激活子(signal transducer and activator of transcription,STAT)等信号通路等途径发挥药效作用,提示保元汤除治疗慢性心力衰竭外,还
基金the Fourth National Investigation of Chinese materia medica resources in Liaoning Province (LN2018017, LN2019019)Career Development Support Plan for Young and Middle-aged Teachers in Shenyang Pharmaceutical University (No. ZQN2021014)Key Laboratory of Marine Biogenetic Resources, Ministry of Natural Resources (No. HY202105)
文摘Objective: To identify phytochemical constituents present in the extract of flowers of Xanthoceras sorbifolia and evaluate their anti-oxidant and anti-hyperglycemic capacities.Methods: The AlCl3colorimetric method and Prussian Blue assay were used to determine the contents of total flavonoids and total phenolic acids in extraction layers, and the bioactive layers was screened through anti-oxidative activity in vitro. The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC BEH C18column(2.0 mm × 150 mm, 5 μm) were used to identify the ingredients. And anti-oxidative ingredients were screened by off-line UPLC-QTOF-MS/MS-free radical scavenging. The ameliorative role of it was further evaluated in a high-fat, streptozotocin-induced type 2 diabetic rat model and the study was carried out on NADPH oxidase(PDB ID: 2CDU) by molecular docking.Results: Combined with the results of activity screening in vitro, the anti-oxidative part was identified as the ethyl acetate layer. A total of 24 chemical constituents were identified by liquid chromatographymass spectrometry in the ethyl acetate layer and 13 main anti-oxidative active constituents were preliminarily screened out through off-line UPLC-QTOF-MS/MS-free radical scavenging. In vivo experiments showed that flowers of X. sorbifolia could significantly reduce the blood glucose level of diabetic mice and alleviate liver cell damage. Based on the results of docking analysis related to the identified phytocompounds and oxidase which involved in type 2 diabetes, quercetin 3-O-rutinoside, kaempferol-3-O-rhamnoside, isorhamnetin-3-O-glucoside, and isoquercitrin showed a better inhibitory profile.Conclusion: The ethyl acetate layer was rich in flavonoids and phenolic acids and had significant anti-oxidant activity, which could prevent hyperglycemia. This observed activity profile suggested X. sorbifolia flowers as a promising new source of tea to develop alternative natural anti-diabetic products with a high safety margin.
