目的在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行...目的在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行时间质谱(UPLC-Q/TOF-MS)联用技术对中药传统水蜜丸制剂二至丸进行多成分定性定量研究的基础上,进行二至丸长期稳定性和加速稳定性研究,并考察了制剂影响因素对制剂稳定性的影响。方法采用Acquity UPLC BEH C18色谱柱(100 mm×2.1mm,1.7μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量0.2 m L/min,采用电喷雾电离源(ESI),正、负离子模式扫描。在遵循ICH制剂稳定性指导原则的指导下,进行了18个月的长期稳定性试验[温度(25±2)℃,相对湿度(RH)(60±5)%],加速稳定性试验[温度(40±2)℃,RH(75±5)%]和制剂影响因素试验(包装材料类型和密封条件)。结果采用以上方法共鉴定出二至丸中20个化学成分(红景天苷、蟛蜞菊内酯、10-羟基木犀榄苷二甲酯、木犀榄苷-11甲酯、马钱苷酸、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷/异毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、特女贞苷异构体、阿富汗丁香苷F、异女贞苷、女贞酸、橄榄苦苷、女贞苷-G13、青藤碱、3-O-顺式-对-香豆酰委陵菜酸),并对其进行定量及归一化半定量研究。在长期稳定性研究中,20个成分均保持稳定;然而在加速稳定性试验条件下,11种成分(10-羟基木犀榄苷二甲酯、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、女贞酸、女贞苷-G13、青藤碱)的量明显下降。制剂影响因素试验结果显示采用钠钙玻璃包装材料替代现有市售聚酯包装,在密封或敞口条件下,制剂中各成分稳定性均显著提高。在采用相同包材时密封条件对制剂稳定性也存在显著性影响。结论通过较为系统的中药传统展开更多
目的 探究蟛蜞菊内酯对对乙酰氨基酚引起的小鼠急性肝损伤的保护作用。方法采用小鼠腹腔注射对乙酰氨基酚模拟肝损伤模型,观察蟛蜞菊内酯对肝的保护作用。将BALB/c小鼠随机分为对照组、模型组、蟛蜞菊内酯低剂量组和高剂量组(n=8)。通...目的 探究蟛蜞菊内酯对对乙酰氨基酚引起的小鼠急性肝损伤的保护作用。方法采用小鼠腹腔注射对乙酰氨基酚模拟肝损伤模型,观察蟛蜞菊内酯对肝的保护作用。将BALB/c小鼠随机分为对照组、模型组、蟛蜞菊内酯低剂量组和高剂量组(n=8)。通过检测小鼠血清中的丙氨酸转移酶(ALT)、天冬氨酸转移酶(AST);肝组织匀浆中的丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-PX)和超氧化物歧化酶(SOD)指标,血清ELIA检测TNF-α、IL-6,结合肝的组织病理学检查,观察蟛蜞菊内酯对对乙酰氨基酚所致肝损伤的保护作用。结果 模型组小鼠血清中的AST、ALT、GSH、MDA、IL-6以及TNF-α水平水平明显高于对照组(P<0.01),有统计学意义;蟛蜞菊内酯低剂量组和高剂量组的AST、ALT以及肝组织匀浆液中SOD和GSH-PX水平明显要高于模型组,且肝组织匀浆液中MDA、GSH水平以及小鼠血清中TNF-α、IL-6水平明显低于模型组(P<0.01),有统计学意义;模型组的GSH-PX和SOD水平明显低于对照组(P<0.01),有统计学意义。组织病理学苏木素-伊红(hematoxylin and eosin,HE)和细胞凋亡染色(TdT-mediated DUTP nick end labeling,TUNEL)显示蟛蜞菊内酯可明显减轻肝组织的坏死和凋亡。结论 蟛蜞菊内酯对对乙酰氨基酚引起的急性肝损伤有明显的保护作用。展开更多
Eclipta prostrata L.has been used in traditional medicine and known for its liver-protective properties for centuries.Wedelolactone(WEL)and demethylwedelolactone(DWEL)are the major coumarins found in E.prostrata L.How...Eclipta prostrata L.has been used in traditional medicine and known for its liver-protective properties for centuries.Wedelolactone(WEL)and demethylwedelolactone(DWEL)are the major coumarins found in E.prostrata L.However,the comprehensive characterization of these two compounds on non-alcoholic fatty liver disease(NAFLD)still remains to be explored.Utilizing a well-established zebrafish model of thioacetamide(TAA)-induced liver injury,the present study sought to investigate the impacts and mechanisms of WEL and DWEL on NAFLD through integrative spatial metabolomics with liver-specific transcriptomics analysis.Our results showed that WEL and DWEL significantly improved liver function and reduced the accumulation of fat in the liver.The biodistributions and metabolism of these two compounds in whole-body zebrafish were successfully mapped,and the discriminatory endogenous metabolites reversely regulated by WEL and DWEL treatments were also characterized.Based on spatial metabolomics and transcriptomics,we identified that steroid biosynthesis and fatty acid metabolism are mainly involved in the hepatoprotective effects of WEL instead of DWEL.Our study unveils the distinct mechanism of WEL and DWEL in ameliorating NAFLD,and presents a“multi-omics”platform of spatial metabolomics and liver-specific transcriptomics to develop highly effective compounds for further improved therapy.展开更多
Objective:To evaluate the effects of wedelolactone,a major flavonoid from Vietnamese Eclipta prostrata(L)L.,on inflammation and insulin resistance.Methods:Wedelolactone was extracted from the leaves of Vietnamese Ecli...Objective:To evaluate the effects of wedelolactone,a major flavonoid from Vietnamese Eclipta prostrata(L)L.,on inflammation and insulin resistance.Methods:Wedelolactone was extracted from the leaves of Vietnamese Eclipta prostrata(L.)L.with methanol by Soxhlet.The effects of wedelolactone on lipopolysaccharide(LPS)-induced cytokine production,reactive oxygen species(ROS)generation,and nicotinamide adenine dinucleotide phosphate(NADPH)oxidase activities in Raw 264.7 cells were measured by enzyme-linked immunosorbent assay(ELISA),specific immunofluorescent dyes and luminometric analysis,respectively.In addition,its effects on glucose uptake and the protein expression of insulin receptor substrate 1(IRS1)and glucose transporter 4(GLUT4)were examined in 3T3-L1 cells by immunofluorescent dyes and Western blot.Results:Wedelolactone at 30μg/mL significantly inhibited LPS-induced production of tumor necrosis factor-α,interleukin(IL)-6,and IL-8(P<0.01)with no noticeable effects on IL-10 level.It also reduced ROS generation and NADPH oxidase activities in LPS-stimulated Raw 264.7 cells(P<0.01).Furthermore,wedelolactone showed anti-insulin resistance activity,as evidenced by improved glucose uptake and the upregulated expression of IRS1 and GLUT4 in 3T3-L1 cells(P<0.01).Conclusions:Wedelolactone exhibits anti-inflammation and anti-insulin resistance effects,which may be used for the treatment of diabetes and inflammation-associated diseases.展开更多
文摘目的在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行时间质谱(UPLC-Q/TOF-MS)联用技术对中药传统水蜜丸制剂二至丸进行多成分定性定量研究的基础上,进行二至丸长期稳定性和加速稳定性研究,并考察了制剂影响因素对制剂稳定性的影响。方法采用Acquity UPLC BEH C18色谱柱(100 mm×2.1mm,1.7μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量0.2 m L/min,采用电喷雾电离源(ESI),正、负离子模式扫描。在遵循ICH制剂稳定性指导原则的指导下,进行了18个月的长期稳定性试验[温度(25±2)℃,相对湿度(RH)(60±5)%],加速稳定性试验[温度(40±2)℃,RH(75±5)%]和制剂影响因素试验(包装材料类型和密封条件)。结果采用以上方法共鉴定出二至丸中20个化学成分(红景天苷、蟛蜞菊内酯、10-羟基木犀榄苷二甲酯、木犀榄苷-11甲酯、马钱苷酸、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷/异毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、特女贞苷异构体、阿富汗丁香苷F、异女贞苷、女贞酸、橄榄苦苷、女贞苷-G13、青藤碱、3-O-顺式-对-香豆酰委陵菜酸),并对其进行定量及归一化半定量研究。在长期稳定性研究中,20个成分均保持稳定;然而在加速稳定性试验条件下,11种成分(10-羟基木犀榄苷二甲酯、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、女贞酸、女贞苷-G13、青藤碱)的量明显下降。制剂影响因素试验结果显示采用钠钙玻璃包装材料替代现有市售聚酯包装,在密封或敞口条件下,制剂中各成分稳定性均显著提高。在采用相同包材时密封条件对制剂稳定性也存在显著性影响。结论通过较为系统的中药传统
文摘目的 探究蟛蜞菊内酯对对乙酰氨基酚引起的小鼠急性肝损伤的保护作用。方法采用小鼠腹腔注射对乙酰氨基酚模拟肝损伤模型,观察蟛蜞菊内酯对肝的保护作用。将BALB/c小鼠随机分为对照组、模型组、蟛蜞菊内酯低剂量组和高剂量组(n=8)。通过检测小鼠血清中的丙氨酸转移酶(ALT)、天冬氨酸转移酶(AST);肝组织匀浆中的丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-PX)和超氧化物歧化酶(SOD)指标,血清ELIA检测TNF-α、IL-6,结合肝的组织病理学检查,观察蟛蜞菊内酯对对乙酰氨基酚所致肝损伤的保护作用。结果 模型组小鼠血清中的AST、ALT、GSH、MDA、IL-6以及TNF-α水平水平明显高于对照组(P<0.01),有统计学意义;蟛蜞菊内酯低剂量组和高剂量组的AST、ALT以及肝组织匀浆液中SOD和GSH-PX水平明显要高于模型组,且肝组织匀浆液中MDA、GSH水平以及小鼠血清中TNF-α、IL-6水平明显低于模型组(P<0.01),有统计学意义;模型组的GSH-PX和SOD水平明显低于对照组(P<0.01),有统计学意义。组织病理学苏木素-伊红(hematoxylin and eosin,HE)和细胞凋亡染色(TdT-mediated DUTP nick end labeling,TUNEL)显示蟛蜞菊内酯可明显减轻肝组织的坏死和凋亡。结论 蟛蜞菊内酯对对乙酰氨基酚引起的急性肝损伤有明显的保护作用。
基金supported by the National Natural Science Foundation of China(Grant No.:82273888)Natural Science Foundation of Shandong Province(Grant Nos.ZR2022QH257,ZR2020YQ60)+2 种基金Shandong Major Technological Innovation Project(Project No.:2021CXGC010508)Taishan Scholars Program of Shandong Province(Program Nos.:tsqn202103096,tsqn202211204)Shandong Province Science and Technology Small and Medium Enterprises Innovation Ability Enhancement Project(Project No.:2022TSGC2210).
文摘Eclipta prostrata L.has been used in traditional medicine and known for its liver-protective properties for centuries.Wedelolactone(WEL)and demethylwedelolactone(DWEL)are the major coumarins found in E.prostrata L.However,the comprehensive characterization of these two compounds on non-alcoholic fatty liver disease(NAFLD)still remains to be explored.Utilizing a well-established zebrafish model of thioacetamide(TAA)-induced liver injury,the present study sought to investigate the impacts and mechanisms of WEL and DWEL on NAFLD through integrative spatial metabolomics with liver-specific transcriptomics analysis.Our results showed that WEL and DWEL significantly improved liver function and reduced the accumulation of fat in the liver.The biodistributions and metabolism of these two compounds in whole-body zebrafish were successfully mapped,and the discriminatory endogenous metabolites reversely regulated by WEL and DWEL treatments were also characterized.Based on spatial metabolomics and transcriptomics,we identified that steroid biosynthesis and fatty acid metabolism are mainly involved in the hepatoprotective effects of WEL instead of DWEL.Our study unveils the distinct mechanism of WEL and DWEL in ameliorating NAFLD,and presents a“multi-omics”platform of spatial metabolomics and liver-specific transcriptomics to develop highly effective compounds for further improved therapy.
基金support from the Fund of The Key Laboratory of Enzyme and Protein Technology,VNU University of Science(KLEPT:22.02).
文摘Objective:To evaluate the effects of wedelolactone,a major flavonoid from Vietnamese Eclipta prostrata(L)L.,on inflammation and insulin resistance.Methods:Wedelolactone was extracted from the leaves of Vietnamese Eclipta prostrata(L.)L.with methanol by Soxhlet.The effects of wedelolactone on lipopolysaccharide(LPS)-induced cytokine production,reactive oxygen species(ROS)generation,and nicotinamide adenine dinucleotide phosphate(NADPH)oxidase activities in Raw 264.7 cells were measured by enzyme-linked immunosorbent assay(ELISA),specific immunofluorescent dyes and luminometric analysis,respectively.In addition,its effects on glucose uptake and the protein expression of insulin receptor substrate 1(IRS1)and glucose transporter 4(GLUT4)were examined in 3T3-L1 cells by immunofluorescent dyes and Western blot.Results:Wedelolactone at 30μg/mL significantly inhibited LPS-induced production of tumor necrosis factor-α,interleukin(IL)-6,and IL-8(P<0.01)with no noticeable effects on IL-10 level.It also reduced ROS generation and NADPH oxidase activities in LPS-stimulated Raw 264.7 cells(P<0.01).Furthermore,wedelolactone showed anti-insulin resistance activity,as evidenced by improved glucose uptake and the upregulated expression of IRS1 and GLUT4 in 3T3-L1 cells(P<0.01).Conclusions:Wedelolactone exhibits anti-inflammation and anti-insulin resistance effects,which may be used for the treatment of diabetes and inflammation-associated diseases.
