Background:Amarogentin(AMA)is a secoiridoid glycoside extracted from Swertia and Gentiana roots and exhibits many biological effects such as antioxidative,antiinflammatory,and antitumor activities.Atopic dermatitis(AD...Background:Amarogentin(AMA)is a secoiridoid glycoside extracted from Swertia and Gentiana roots and exhibits many biological effects such as antioxidative,antiinflammatory,and antitumor activities.Atopic dermatitis(AD)is a chronic inflammatory skin disease caused by disorders in the regulation of multiple inflammatory cytokines.No effective cure has been found for AD now.Methods:We constructed the HaCat and splenocyte model and tested the inhibitory effect of AMA on IL-4,IL-6,and IL-13 secretions using enzyme-linked immunosorbent assay(ELISA).The AD mouse model was constructed and treated with AMA,the severity of skin lesions was observed,epidermal tissue was collected,and epidermal thickness and mast cell infiltration were observed using hematoxylin and eosin and toluidine blue staining,respectively.The expression of kallikrein-related peptidase 7(KLK7)and filaggrin(FLG)was detected using immunostaining and Western blot analysis.The mRNA expression of KLK7 and FLG was detected using quantitative polymerase chain reaction(qPCR).Blood immunoglobulin E(IgE)secretion was detected.Results:AMA inhibited IL-6 secreted by tumor necrosis factor(TNF)-α-induced HaCaT cells and reduced IL-4 and IL-13 secreted by phytohemagglutinin(PHA)-induced primary cells in the mice spleen.It was found that the treatment of AMA with 2,4-din itrochlorobenzene-induced AD-like mice could promote the recovery of dermatitis,reduce the score of dermatitis severity and the scratching frequency,treat the skin lesions,reduce the epidermal thickness,decrease the infiltration of mast cells,reduce the IgE level in serum,decrease the expression levels of AD-related cytokines,increase protein and mRNA expression of FLG,and reduce the protein and mRNA expression of KLK7 in the skin tissues of AD-like mice.Conclusion:In conclusion,AMA inhibits inflammatory response at the cellular level,and AMA reduces the validation response of specific dermatitis mice,relieves pruritus,and repairs the damaged skin barrier.展开更多
目的:建立抱茎獐牙菜指纹图谱及多指标成分含量测定方法,为其质量控制及评价提供参考。方法:采用Waters Sun Fir ODS C18色谱柱,甲醇-0.05%磷酸溶液为流动相梯度洗脱,体积流量为1.0 m L·min^(-1),检测波长为254 nm,柱温为30℃;建...目的:建立抱茎獐牙菜指纹图谱及多指标成分含量测定方法,为其质量控制及评价提供参考。方法:采用Waters Sun Fir ODS C18色谱柱,甲醇-0.05%磷酸溶液为流动相梯度洗脱,体积流量为1.0 m L·min^(-1),检测波长为254 nm,柱温为30℃;建立高效液相色谱(HPLC)图谱,采用Chempattern化学计量学软件进行相似度分析。进一步通过HPLC法,测定样品中10个指标成分的含量,以寻找样品间的质量差异成分。结果:建立了抱茎獐牙菜HPLC指纹图谱,相似度均>0.904,并确认29个共有峰,指认出马钱苷酸、獐牙菜苦苷、龙胆苦苷、当药苷、芒果苷、异荭草素、苦龙胆酯苷、当药醇苷、1,8-二羟基-3,7-二甲氧基[口山]酮、雏菊叶龙胆酮10个色谱峰。含量测定显示,10个成分线性关系良好(r≥0.9993),加样回收率均在96.8%~99.3%,RSD<2.1%,平均含量分别为0.16%、3.63%、0.23%、0.38%、0.24%、0.26%、0.07%、1.13%、0.08%、0.25%。测定结果表明獐牙菜苦苷、当药醇苷等成分含量较高。结论:HPLC指纹图谱结合多指标成分含量测定方法适用于抱茎獐牙菜的质量控制与评价,该方法快捷简便、稳定可靠,可为建立藏药抱茎獐牙菜质量标准提供依据。展开更多
Amarogentin is an efficacious Chinese herbal medicine and a component of the bitter apricot kernel.It is commonly used as an expectorant and supplementary anti-cancer drug.β-Glucosidase is an enzyme that hydrolyzes t...Amarogentin is an efficacious Chinese herbal medicine and a component of the bitter apricot kernel.It is commonly used as an expectorant and supplementary anti-cancer drug.β-Glucosidase is an enzyme that hydrolyzes the glycosidic bond between aryl and saccharide groups to release glucose.Upon their interaction,β-glucosidase catalyzes amarogentin to produce considerable amounts of hydrocyanic acid,which inhibits cytochrome C oxidase,the terminal enzyme in the mitochondrial respiration chain,and suspends adenosine triphosphate synthesis,resulting in cell death.Hydrocyanic acid is a cell-cycle-stage-nonspecific agent that kills cancer cells.Thus,β-glucosidase can be coupled with a tumor-specific monoclonal antibody.β-Glucosidase can combine with cancer-cell-surface antigens and specifically convert amarogentin to an active drug that acts on cancer cells and the surrounding antibodies to achieve a kil ing effect.β-Glucosidase is injected intravenously and recognizes cancer-cel-surface antigens with the help of an antibody.The prodrug amarogentin is infused after β-glucosidase has reached the target position.Coupling of cell membrane peptides with β-glucosidase allows the enzyme to penetrate capillary endothelial cells and clear extracellular deep solid tumors to kill the cells therein.The Chinese medicine amarogentin and β-glucosidase will become an important treatment for various tumors when an appropriate monoclonal antibody is developed.展开更多
基金The present study was supported by the National Natural Science Foundation of China(grant numbers:81902067 and 82078189)the Medical Scientific Research Foundation of Guangdong Province(grant number:A2019502)+2 种基金the Shenzhen Science and Technology Innovation Committee(grant numbers:JCY20180305124849781 and 20200812211704001)the SZU Top Ranking Project(grant number:86000000210)the Hospital Project of Huazhong University of Science and Technology Union Shenzhen Hospital(grant number:2021042).
文摘Background:Amarogentin(AMA)is a secoiridoid glycoside extracted from Swertia and Gentiana roots and exhibits many biological effects such as antioxidative,antiinflammatory,and antitumor activities.Atopic dermatitis(AD)is a chronic inflammatory skin disease caused by disorders in the regulation of multiple inflammatory cytokines.No effective cure has been found for AD now.Methods:We constructed the HaCat and splenocyte model and tested the inhibitory effect of AMA on IL-4,IL-6,and IL-13 secretions using enzyme-linked immunosorbent assay(ELISA).The AD mouse model was constructed and treated with AMA,the severity of skin lesions was observed,epidermal tissue was collected,and epidermal thickness and mast cell infiltration were observed using hematoxylin and eosin and toluidine blue staining,respectively.The expression of kallikrein-related peptidase 7(KLK7)and filaggrin(FLG)was detected using immunostaining and Western blot analysis.The mRNA expression of KLK7 and FLG was detected using quantitative polymerase chain reaction(qPCR).Blood immunoglobulin E(IgE)secretion was detected.Results:AMA inhibited IL-6 secreted by tumor necrosis factor(TNF)-α-induced HaCaT cells and reduced IL-4 and IL-13 secreted by phytohemagglutinin(PHA)-induced primary cells in the mice spleen.It was found that the treatment of AMA with 2,4-din itrochlorobenzene-induced AD-like mice could promote the recovery of dermatitis,reduce the score of dermatitis severity and the scratching frequency,treat the skin lesions,reduce the epidermal thickness,decrease the infiltration of mast cells,reduce the IgE level in serum,decrease the expression levels of AD-related cytokines,increase protein and mRNA expression of FLG,and reduce the protein and mRNA expression of KLK7 in the skin tissues of AD-like mice.Conclusion:In conclusion,AMA inhibits inflammatory response at the cellular level,and AMA reduces the validation response of specific dermatitis mice,relieves pruritus,and repairs the damaged skin barrier.
文摘目的:建立抱茎獐牙菜指纹图谱及多指标成分含量测定方法,为其质量控制及评价提供参考。方法:采用Waters Sun Fir ODS C18色谱柱,甲醇-0.05%磷酸溶液为流动相梯度洗脱,体积流量为1.0 m L·min^(-1),检测波长为254 nm,柱温为30℃;建立高效液相色谱(HPLC)图谱,采用Chempattern化学计量学软件进行相似度分析。进一步通过HPLC法,测定样品中10个指标成分的含量,以寻找样品间的质量差异成分。结果:建立了抱茎獐牙菜HPLC指纹图谱,相似度均>0.904,并确认29个共有峰,指认出马钱苷酸、獐牙菜苦苷、龙胆苦苷、当药苷、芒果苷、异荭草素、苦龙胆酯苷、当药醇苷、1,8-二羟基-3,7-二甲氧基[口山]酮、雏菊叶龙胆酮10个色谱峰。含量测定显示,10个成分线性关系良好(r≥0.9993),加样回收率均在96.8%~99.3%,RSD<2.1%,平均含量分别为0.16%、3.63%、0.23%、0.38%、0.24%、0.26%、0.07%、1.13%、0.08%、0.25%。测定结果表明獐牙菜苦苷、当药醇苷等成分含量较高。结论:HPLC指纹图谱结合多指标成分含量测定方法适用于抱茎獐牙菜的质量控制与评价,该方法快捷简便、稳定可靠,可为建立藏药抱茎獐牙菜质量标准提供依据。
文摘Amarogentin is an efficacious Chinese herbal medicine and a component of the bitter apricot kernel.It is commonly used as an expectorant and supplementary anti-cancer drug.β-Glucosidase is an enzyme that hydrolyzes the glycosidic bond between aryl and saccharide groups to release glucose.Upon their interaction,β-glucosidase catalyzes amarogentin to produce considerable amounts of hydrocyanic acid,which inhibits cytochrome C oxidase,the terminal enzyme in the mitochondrial respiration chain,and suspends adenosine triphosphate synthesis,resulting in cell death.Hydrocyanic acid is a cell-cycle-stage-nonspecific agent that kills cancer cells.Thus,β-glucosidase can be coupled with a tumor-specific monoclonal antibody.β-Glucosidase can combine with cancer-cell-surface antigens and specifically convert amarogentin to an active drug that acts on cancer cells and the surrounding antibodies to achieve a kil ing effect.β-Glucosidase is injected intravenously and recognizes cancer-cel-surface antigens with the help of an antibody.The prodrug amarogentin is infused after β-glucosidase has reached the target position.Coupling of cell membrane peptides with β-glucosidase allows the enzyme to penetrate capillary endothelial cells and clear extracellular deep solid tumors to kill the cells therein.The Chinese medicine amarogentin and β-glucosidase will become an important treatment for various tumors when an appropriate monoclonal antibody is developed.
