This study was designed to determine the impact of chrysoeriol on proliferation and cell cycle progression in the human multiple myeloma cell lines RPMI 8226 and KM3,and its related molecular mechanisms.Chryseoriol wa...This study was designed to determine the impact of chrysoeriol on proliferation and cell cycle progression in the human multiple myeloma cell lines RPMI 8226 and KM3,and its related molecular mechanisms.Chryseoriol was identified by using the phosphorylated AKT-specific cytoblot high throughput assay.CCK-8 assay was employed to examine the growth inhibition rate and IC 50 (48 h) in peripheral blood mononuclear cells (PBMNCs),RPMI 8226 and KM3 cells treated with chrysoeriol at various concentrations.Cells were labeled with 5-6-carboxyfluorescein diacetate succinimidyl ester (CFSE),and the proliferation dynamics was detected by flow cytometry and analyzed with ModFit software.The cell cycles of RPMI 8226 and KM3 cells were measured by flow cytometry when the IC 50 concentration of chrysoeriol was adopted.The alterations in cell-cycle related proteins (Cyclin B1,Cyclin D1,p21) and proteins in PI3K-AKT-mTOR pathway were determined by Western blot analysis.The results showed the proliferation of multiple myeloma cells was significantly inhibited by chrysoeriol,resulting in cell cycle arrest in G 2 /M phase.Chrysoeriol could significantly reduce the expression of p-AKT (s473) and p-4eBP1 (t37/46) protein,meanwhile enhanced Cyclin B1 and p21 protein expression.Similar effects were not observed in PBMNCs from normal donors.It was concluded that chrysoeriol was a selective PI3K-AKT-mTOR pathway inhibitor.It restrained the proliferation of human multiple myeloma cells,but didn’t affect proliferation of PBMNCs from normal donors.It might exhibit the cell cycle regulatory effect via the inhibition of PI3K-AKT-mTOR signal pathway.展开更多
The present study aimed to evaluate the effects of chrysoeriol from Cardiospermum halicacabum in streptozotocin induced Wistar rats.Thirty rats were categorized as control,diabetic control supplemented with 0,20 mg/kg...The present study aimed to evaluate the effects of chrysoeriol from Cardiospermum halicacabum in streptozotocin induced Wistar rats.Thirty rats were categorized as control,diabetic control supplemented with 0,20 mg/kg chrysoeriol and 600μg/kg BW of glibenclamide for 45-day trial period.Our results indicated that the inclusion of chrysoeriol(20 mg/kg)showed a significant reduction in plasma glucose,hemoglobin and glycosylated hemoglobin level with a rising of plasma insulin sensitivity.Further,downregulated enzymes including glucose 6-phosphatase,fructose 1,6-bisphosphatase,and glycogen phosphorylase as well upregulated enzymes such as hexokinase,glucose-6-phosphate dehydrogenase,pyruvate kinase,and hepatic glycogen content.There was a diminish action found in liver glycogen synthase of tested rat with a rise in gamma-glutamyl transpeptidase,towards normal levels upon treatment with chrysoeriol.The histopathological study confirmed that renewal of the beta cells of pancreatic of chrysoeriol and glibenclamide treated rats.In addition,the molecular docking of chrysoeriol against glycolytic enzymes including hexokinase,glucose-6-phosphate dehydrogenase,pyruvate kinase,using Argus software shows chrysoeriol had greatest ligand binding energy as equivalent to glibenclamide,as a standard drug.Thus,chrysoeriol found to be non-toxic with potential regulation on glycemic control and upregulation of the carbohydrate metabolic enzymes.展开更多
Background The use of doxorubicin （DOX） is limited by its dose-dependent cardiotoxicity. Reactive oxygen species （ROSs） play an important role in the pathological process of DOX-induced cardiotoxicity. The aim of ...Background The use of doxorubicin （DOX） is limited by its dose-dependent cardiotoxicity. Reactive oxygen species （ROSs） play an important role in the pathological process of DOX-induced cardiotoxicity. The aim of this study was to evaluate the protective effect of chrysoeriol, a flavone compound, against DOX-induced apoptosis and death in H9c2 cells and to find out its preliminary mechanism. Methods We used 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl-2H-tetrazolium bromide （MTT） assay, Hoechst33258 staining and measurement of lactate dehydrogenase （LDH） release to evaluate the protective effect of chrysoeriol against DOX-induced apoptosis and death in H9c2 cells. To find out the mechanism of this protective effect, we observed the immunofluorescence of intracellular ROS and measured the activities of malondialdehyde （MDA）, superoxide dismutase （SOD） and glutathione peroxidase （GPx）. Furthermore, we evaluated the effect of chrysoeriol on the antitumor activity of DOX in HeLa cells with MTT assay. Results The results of MTT assay, Hoechst 33258 staining and measurement of LDH release showed that chrysoeriol significantly reduced doxorubicin-induced apoptosis and cell death. Chrysoeriol at a dose of 20 μg/ml notably reduced intracellular ROS, decreased the concentration of MDA in the supernatant of DOX-treated H9c2 cells and increased SOD and GPx activities to their normal levels. Further study showed that the addition of chrysoeriol did not affect the antitumor activity of DOX. Conclusion Chrysoeriol could potentially serve as a novel cardioprotective agent against DOX-induced cardiotoxicity without affecting the antitumor activity of DOX.展开更多
目的:采用网络药理学、分子对接技术及细胞实验等方法,系统探究花生衣中金圣草黄素通过磷脂酰肌醇3-激酶-蛋白激酶B-哺乳动物雷帕霉素靶蛋白(phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin,PI3K-AKT...目的:采用网络药理学、分子对接技术及细胞实验等方法,系统探究花生衣中金圣草黄素通过磷脂酰肌醇3-激酶-蛋白激酶B-哺乳动物雷帕霉素靶蛋白(phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin,PI3K-AKT-mTOR)通路抑制肝癌的作用机制。方法:利用传统中药系统药理学(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库筛选出花生衣的活性成分,预测肝癌相关靶点,通过基因本体(gene ontology,GO)功能与京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析评估关键靶点涉及的生物过程与信号通路。采用分子对接解析金圣草黄素与PI3K、AKT、mTOR、固醇调节元件结合蛋白1(sterol regulatory element-binding protein 1,SREBP-1)、脂肪酸合酶(fatty acid synthase,FASN)和硬脂酰辅酶A去饱和酶1(stearoyl-CoA desaturase 1,SCD1)之间的相互作用。通过细胞计数试剂盒-8(Cell Counting Kit-8,CCK-8)进行细胞增殖实验、克隆形成实验及蛋白质印迹(Western blot)分析,验证金圣草黄素对肝细胞癌细胞系G2细胞(hepatocellular carcinoma G2,HepG2)增殖及脂肪酸合成的抑制作用。结果:网络药理学分析筛选出了花生衣的关键靶点,并揭示了PI3K-AKT-mTOR信号通路在金圣草黄素抗肝癌作用中的核心调控地位。GO功能富集分析显示,金圣草黄素主要涉及细胞凋亡、外源性刺激应答及类固醇激素反应等生物过程。KEGG通路富集分析进一步证实PI3K-AKT通路在其抗肝癌中的关键作用。分子对接结果表明,金圣草黄素可与丝氨酸/苏氨酸激酶1、mTOR、SREBP-1、FASN和SCD1形成稳定结合,其结合自由能均<-5 kcal/mol,揭示其具有较高的结合亲和力。细胞实验结果显示,金圣草黄素能够显著抑制HepG2细胞的增殖,并通过下调磷酸化AKT(p-AKT)、mTOR、SREBP-1、FASN和SCD1的蛋白表达,抑制肝癌细胞的脂肪酸合成,进一步验证了其在PI3K-AKT-mTOR信号通路中的调控作用。结论:金圣草黄素通过调控PI3K-AKT-mTOR信号通路,抑制肝癌细胞的增殖与脂质代谢,揭示了其作为花生衣中潜在抗肝癌成分的药理机制。展开更多
目的研究柯伊利素对痛风性关节炎小鼠的治疗作用及其免疫机制。方法18只C57BL/6小鼠随机分为假手术(sham)组、模型(model)组及柯伊利素治疗组,每组6只,采用微晶尿酸钠(MSU)注射小鼠后肢踝关节造模成痛风性关节炎小鼠模型;每日给予柯伊利...目的研究柯伊利素对痛风性关节炎小鼠的治疗作用及其免疫机制。方法18只C57BL/6小鼠随机分为假手术(sham)组、模型(model)组及柯伊利素治疗组,每组6只,采用微晶尿酸钠(MSU)注射小鼠后肢踝关节造模成痛风性关节炎小鼠模型;每日给予柯伊利素200 mg/kg注射治疗7 d。观察关节肿胀程度,记录后肢功能障碍指数;HE染色观察关节滑膜组织病理变化;ELISA检测血清中尿素(uric acid,UA)含量及关节组织液中IL-1β、IL-6、IL-10、IL-18和TNF-α等炎症因子的水平;qRTPCR和Western blotting分别检测NF-κB、NLR家族热蛋白结构域包含蛋白3(NLR family pyrin domain containing 3,NLPR3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)和Caspase-1的mRNA和蛋白水平;流式细胞术检测组织液中Th17和Treg细胞数量。结果与模型组相比,柯伊利素有效降低了小鼠关节肿胀程度和后肢关节障碍指数,改善了关节滑膜液组织病理恶化;血清中UA含量下降,关节液中的炎症因子如IL-1β、IL-6、IL-18和TNF-α等含量明显减少,IL-10含量增加;另外,qRT-PCR结果显示,NF-κB、NLPR3、ASC和Caspase-1的mRNA表达水平下降,且NLPR3、ASC和Caspase-1的蛋白表达趋势与mRNA表达趋势一致;流式细胞术检测发现,Th17细胞频率和Th17/Treg与模型组比较均下降,而Treg细胞频率升高。结论柯伊利素能有效缓解小鼠痛风性关节炎的症状,可能是通过影响NF-κB-NLRP3信号通路,调控炎性相关细胞子的表达,并通过改善Th17/Treg平衡减轻组织局部的炎症。展开更多
Genus Codonopsis comprises more than 40 species in China. The roots of these plants have been used as traditional Chinese medicines or folk love herbs. Codonopsis xundianensis Wang ZT et Xu GJ grows in Yunnan Province...Genus Codonopsis comprises more than 40 species in China. The roots of these plants have been used as traditional Chinese medicines or folk love herbs. Codonopsis xundianensis Wang ZT et Xu GJ grows in Yunnan Province and its roots are often used as a tonic. In this paper, we report the isolation and structural elucidation of chrysoeriol (1), tricin (2), wogonin (3) and luteolin (4) from the roots of Codonopsis xundianensis. Compounds 1-3 were found in Codonopsis for the first time.展开更多
基金supported by grants from the National Natural Sciences Foundation of China(No.30770914No.30901587)China State Key Basic Research Program(No.2002CB513100)
文摘This study was designed to determine the impact of chrysoeriol on proliferation and cell cycle progression in the human multiple myeloma cell lines RPMI 8226 and KM3,and its related molecular mechanisms.Chryseoriol was identified by using the phosphorylated AKT-specific cytoblot high throughput assay.CCK-8 assay was employed to examine the growth inhibition rate and IC 50 (48 h) in peripheral blood mononuclear cells (PBMNCs),RPMI 8226 and KM3 cells treated with chrysoeriol at various concentrations.Cells were labeled with 5-6-carboxyfluorescein diacetate succinimidyl ester (CFSE),and the proliferation dynamics was detected by flow cytometry and analyzed with ModFit software.The cell cycles of RPMI 8226 and KM3 cells were measured by flow cytometry when the IC 50 concentration of chrysoeriol was adopted.The alterations in cell-cycle related proteins (Cyclin B1,Cyclin D1,p21) and proteins in PI3K-AKT-mTOR pathway were determined by Western blot analysis.The results showed the proliferation of multiple myeloma cells was significantly inhibited by chrysoeriol,resulting in cell cycle arrest in G 2 /M phase.Chrysoeriol could significantly reduce the expression of p-AKT (s473) and p-4eBP1 (t37/46) protein,meanwhile enhanced Cyclin B1 and p21 protein expression.Similar effects were not observed in PBMNCs from normal donors.It was concluded that chrysoeriol was a selective PI3K-AKT-mTOR pathway inhibitor.It restrained the proliferation of human multiple myeloma cells,but didn’t affect proliferation of PBMNCs from normal donors.It might exhibit the cell cycle regulatory effect via the inhibition of PI3K-AKT-mTOR signal pathway.
文摘The present study aimed to evaluate the effects of chrysoeriol from Cardiospermum halicacabum in streptozotocin induced Wistar rats.Thirty rats were categorized as control,diabetic control supplemented with 0,20 mg/kg chrysoeriol and 600μg/kg BW of glibenclamide for 45-day trial period.Our results indicated that the inclusion of chrysoeriol(20 mg/kg)showed a significant reduction in plasma glucose,hemoglobin and glycosylated hemoglobin level with a rising of plasma insulin sensitivity.Further,downregulated enzymes including glucose 6-phosphatase,fructose 1,6-bisphosphatase,and glycogen phosphorylase as well upregulated enzymes such as hexokinase,glucose-6-phosphate dehydrogenase,pyruvate kinase,and hepatic glycogen content.There was a diminish action found in liver glycogen synthase of tested rat with a rise in gamma-glutamyl transpeptidase,towards normal levels upon treatment with chrysoeriol.The histopathological study confirmed that renewal of the beta cells of pancreatic of chrysoeriol and glibenclamide treated rats.In addition,the molecular docking of chrysoeriol against glycolytic enzymes including hexokinase,glucose-6-phosphate dehydrogenase,pyruvate kinase,using Argus software shows chrysoeriol had greatest ligand binding energy as equivalent to glibenclamide,as a standard drug.Thus,chrysoeriol found to be non-toxic with potential regulation on glycemic control and upregulation of the carbohydrate metabolic enzymes.
基金This work was supported by grants from the Ministry of Science and Technology, China （No. 2006DFA31500） and Natural Science Foundation of China （No. 30640080）.
文摘Background The use of doxorubicin （DOX） is limited by its dose-dependent cardiotoxicity. Reactive oxygen species （ROSs） play an important role in the pathological process of DOX-induced cardiotoxicity. The aim of this study was to evaluate the protective effect of chrysoeriol, a flavone compound, against DOX-induced apoptosis and death in H9c2 cells and to find out its preliminary mechanism. Methods We used 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl-2H-tetrazolium bromide （MTT） assay, Hoechst33258 staining and measurement of lactate dehydrogenase （LDH） release to evaluate the protective effect of chrysoeriol against DOX-induced apoptosis and death in H9c2 cells. To find out the mechanism of this protective effect, we observed the immunofluorescence of intracellular ROS and measured the activities of malondialdehyde （MDA）, superoxide dismutase （SOD） and glutathione peroxidase （GPx）. Furthermore, we evaluated the effect of chrysoeriol on the antitumor activity of DOX in HeLa cells with MTT assay. Results The results of MTT assay, Hoechst 33258 staining and measurement of LDH release showed that chrysoeriol significantly reduced doxorubicin-induced apoptosis and cell death. Chrysoeriol at a dose of 20 μg/ml notably reduced intracellular ROS, decreased the concentration of MDA in the supernatant of DOX-treated H9c2 cells and increased SOD and GPx activities to their normal levels. Further study showed that the addition of chrysoeriol did not affect the antitumor activity of DOX. Conclusion Chrysoeriol could potentially serve as a novel cardioprotective agent against DOX-induced cardiotoxicity without affecting the antitumor activity of DOX.
文摘目的:采用网络药理学、分子对接技术及细胞实验等方法,系统探究花生衣中金圣草黄素通过磷脂酰肌醇3-激酶-蛋白激酶B-哺乳动物雷帕霉素靶蛋白(phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin,PI3K-AKT-mTOR)通路抑制肝癌的作用机制。方法:利用传统中药系统药理学(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库筛选出花生衣的活性成分,预测肝癌相关靶点,通过基因本体(gene ontology,GO)功能与京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析评估关键靶点涉及的生物过程与信号通路。采用分子对接解析金圣草黄素与PI3K、AKT、mTOR、固醇调节元件结合蛋白1(sterol regulatory element-binding protein 1,SREBP-1)、脂肪酸合酶(fatty acid synthase,FASN)和硬脂酰辅酶A去饱和酶1(stearoyl-CoA desaturase 1,SCD1)之间的相互作用。通过细胞计数试剂盒-8(Cell Counting Kit-8,CCK-8)进行细胞增殖实验、克隆形成实验及蛋白质印迹(Western blot)分析,验证金圣草黄素对肝细胞癌细胞系G2细胞(hepatocellular carcinoma G2,HepG2)增殖及脂肪酸合成的抑制作用。结果:网络药理学分析筛选出了花生衣的关键靶点,并揭示了PI3K-AKT-mTOR信号通路在金圣草黄素抗肝癌作用中的核心调控地位。GO功能富集分析显示,金圣草黄素主要涉及细胞凋亡、外源性刺激应答及类固醇激素反应等生物过程。KEGG通路富集分析进一步证实PI3K-AKT通路在其抗肝癌中的关键作用。分子对接结果表明,金圣草黄素可与丝氨酸/苏氨酸激酶1、mTOR、SREBP-1、FASN和SCD1形成稳定结合,其结合自由能均<-5 kcal/mol,揭示其具有较高的结合亲和力。细胞实验结果显示,金圣草黄素能够显著抑制HepG2细胞的增殖,并通过下调磷酸化AKT(p-AKT)、mTOR、SREBP-1、FASN和SCD1的蛋白表达,抑制肝癌细胞的脂肪酸合成,进一步验证了其在PI3K-AKT-mTOR信号通路中的调控作用。结论:金圣草黄素通过调控PI3K-AKT-mTOR信号通路,抑制肝癌细胞的增殖与脂质代谢,揭示了其作为花生衣中潜在抗肝癌成分的药理机制。
文摘目的研究柯伊利素对痛风性关节炎小鼠的治疗作用及其免疫机制。方法18只C57BL/6小鼠随机分为假手术(sham)组、模型(model)组及柯伊利素治疗组,每组6只,采用微晶尿酸钠(MSU)注射小鼠后肢踝关节造模成痛风性关节炎小鼠模型;每日给予柯伊利素200 mg/kg注射治疗7 d。观察关节肿胀程度,记录后肢功能障碍指数;HE染色观察关节滑膜组织病理变化;ELISA检测血清中尿素(uric acid,UA)含量及关节组织液中IL-1β、IL-6、IL-10、IL-18和TNF-α等炎症因子的水平;qRTPCR和Western blotting分别检测NF-κB、NLR家族热蛋白结构域包含蛋白3(NLR family pyrin domain containing 3,NLPR3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)和Caspase-1的mRNA和蛋白水平;流式细胞术检测组织液中Th17和Treg细胞数量。结果与模型组相比,柯伊利素有效降低了小鼠关节肿胀程度和后肢关节障碍指数,改善了关节滑膜液组织病理恶化;血清中UA含量下降,关节液中的炎症因子如IL-1β、IL-6、IL-18和TNF-α等含量明显减少,IL-10含量增加;另外,qRT-PCR结果显示,NF-κB、NLPR3、ASC和Caspase-1的mRNA表达水平下降,且NLPR3、ASC和Caspase-1的蛋白表达趋势与mRNA表达趋势一致;流式细胞术检测发现,Th17细胞频率和Th17/Treg与模型组比较均下降,而Treg细胞频率升高。结论柯伊利素能有效缓解小鼠痛风性关节炎的症状,可能是通过影响NF-κB-NLRP3信号通路,调控炎性相关细胞子的表达,并通过改善Th17/Treg平衡减轻组织局部的炎症。
文摘Genus Codonopsis comprises more than 40 species in China. The roots of these plants have been used as traditional Chinese medicines or folk love herbs. Codonopsis xundianensis Wang ZT et Xu GJ grows in Yunnan Province and its roots are often used as a tonic. In this paper, we report the isolation and structural elucidation of chrysoeriol (1), tricin (2), wogonin (3) and luteolin (4) from the roots of Codonopsis xundianensis. Compounds 1-3 were found in Codonopsis for the first time.
