Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Metho...Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Methods:L.erythrorhizon was extracted by ultrasonication in 95%ethanol,and determined using high-performance liquid chromatography(HPLC).He La,A549,SW620,HEK-293 T,THP-1,K562,Raw264.7 and MDA-MB-231 cell lines were used to investigate the morphology and mechanism of pyroptosis induced by L.erythrorhizon.The lactate dehydrogenase(LDH)release,propidium iodide(PI)/Hoechst double-staining,and pyroptosis reconstitution experiments were performed to study L.erythrorhizon-induced cell pyroptosis.Results:Compared with the death inhibitor,PI/Hoechst and LDH release experiments,we found that L.erythrorhizon induced pyroptosis.Recombination and western blot experiments confired that L.erythrorhizon induced GSDME cleavage,which drives pyroptosis.This phenomenon is conserved in several cancer cell lines that might be triggered by caspase family proteases.The mechanism of L.erythrorhizon inducing pyroptosis is widely found in tumor cells.Conclusion:Our findings not only explain how L.erythrorhizon triggers cancer cell pyroptosis,but also provide mechanistic insights to guide its clinical application in the future.展开更多
In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized ...In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized to determine the nephrotoxicity of these fractions with the help of fluorescence image analysis.As a result,the primary fraction A05 and its secondary fractions C06 ＂C09 and C12 ＂C14 were found to have significant toxicity to LLC-PK1 cell line,as determined by the survive rate less than 20% after they were treated with these fractions.These potential nephrotoxic fractions were further analyzed by multistage and high resolution mass spectrometry.The main compounds in these fractions were tentatively identified to be acetylshikonin,isobutyrylshikonin,β,β＇-dimethyla-cryloylshikonin,and isovalerylshikonin,which may bring nephrotoxicity.展开更多
[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used t...[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used to evaluate the hair coloring effect and the absorbance of cleaning solution after hair was dyed. [Results] The optimum pet dog dyeing process of plant hair dye was dye concentration of50 mg/mL,dye pH of 7. 5 and dyeing time of 1 h. [Conclusions] The coloring and fixing effects of the dye and dyeing process are good,which can lay a foundation for the development and utilization of plant dyes.展开更多
We have developed an on-line detection method using acridine orange as the fluorescence probe and applied this method to rapidly identify active compounds in herbal medicines. This on-line method was equipped with a h...We have developed an on-line detection method using acridine orange as the fluorescence probe and applied this method to rapidly identify active compounds in herbal medicines. This on-line method was equipped with a high-performance liquid chromatography tandem diode array detector, electrospray ionization-ion-trap time-of-flight mass spectrometry and DNA- acridine orange fluorescence detection （HPLC-DAD-MSn-DNA-AO-FLD）. A large amount of information could be simultaneously obtained during one run, which included HPLC fingerprint, ultraviolet spectra, total ion chromatograms, MSn data of high-resolution mass spectrometry and activity profile of each compound binding with DNA. The method also provided information on structureactivity relationships and mechanism of interaction. We used this on-line method to identify five DNA-binding activity components from Lithospermum erythrorhizon sample for the first time. The result showed that the parent nucleus of shikonin derivatives could bind with DNA. The structure-activity relationship showed that the parent nucleus of shikonin derivatives plays a major role in DNA binding, not the carboxyl group on the side chain. This simple, rapid, high precision and good stability on-line method should be useful for compound separation, structural identification and screening of DNA-binding compounds in herbal medicines.展开更多
Crown gall tissue culture lines of Lithospermum erythrorhizon were established by transformation of sterile seedlings with Agrobacterium tumefaciens C58. Nopaline was inspected in the crown gall and showed that the tr...Crown gall tissue culture lines of Lithospermum erythrorhizon were established by transformation of sterile seedlings with Agrobacterium tumefaciens C58. Nopaline was inspected in the crown gall and showed that the transformation was successfull. L6 line, can secret red naphthoquinone pigments that are shikonin derivatives on phytohormone free 6,7-V medium, was selected from other lines on different basic phytohormone free media(MS, 6,7-V, WP, B5).The content of shikonin in the crown gall was 1 18%(dry tissue).The study showed that crown gall tissue culture of L.erythrorhizon was a new method to produce shikonin.展开更多
基金supported by Program for the research startup fund program at Beijing University of Chinese Medicine(90011451310011)the key research fund for drug discovery in Chinese medicine at Beijing University of Chinese Medicine(1000061223740)+1 种基金the experimental technology standardization research project at Beijing University of Chinese Medicine(2021-SYJS-009)the fundamental research funds for the central universities of Beijing University of Chinese Medicine(2020-JYBZDGG-057)。
文摘Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Methods:L.erythrorhizon was extracted by ultrasonication in 95%ethanol,and determined using high-performance liquid chromatography(HPLC).He La,A549,SW620,HEK-293 T,THP-1,K562,Raw264.7 and MDA-MB-231 cell lines were used to investigate the morphology and mechanism of pyroptosis induced by L.erythrorhizon.The lactate dehydrogenase(LDH)release,propidium iodide(PI)/Hoechst double-staining,and pyroptosis reconstitution experiments were performed to study L.erythrorhizon-induced cell pyroptosis.Results:Compared with the death inhibitor,PI/Hoechst and LDH release experiments,we found that L.erythrorhizon induced pyroptosis.Recombination and western blot experiments confired that L.erythrorhizon induced GSDME cleavage,which drives pyroptosis.This phenomenon is conserved in several cancer cell lines that might be triggered by caspase family proteases.The mechanism of L.erythrorhizon inducing pyroptosis is widely found in tumor cells.Conclusion:Our findings not only explain how L.erythrorhizon triggers cancer cell pyroptosis,but also provide mechanistic insights to guide its clinical application in the future.
基金Supported by the National Key Scientific and Technological Project of China(No.2009ZX09502-012)the Research Fund for the Doctoral Program of Higher Education of China(No.20090101110126)the Zhejiang Province Science and Technology Plan Project,China(No.2008C23065)
文摘In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized to determine the nephrotoxicity of these fractions with the help of fluorescence image analysis.As a result,the primary fraction A05 and its secondary fractions C06 ＂C09 and C12 ＂C14 were found to have significant toxicity to LLC-PK1 cell line,as determined by the survive rate less than 20% after they were treated with these fractions.These potential nephrotoxic fractions were further analyzed by multistage and high resolution mass spectrometry.The main compounds in these fractions were tentatively identified to be acetylshikonin,isobutyrylshikonin,β,β＇-dimethyla-cryloylshikonin,and isovalerylshikonin,which may bring nephrotoxicity.
基金Supported by the National Science and Technology Foundation Project of Traditional Chinese Veterinary Medicine Resources Rescuing and Organizing(2013FY110600-03)
文摘[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used to evaluate the hair coloring effect and the absorbance of cleaning solution after hair was dyed. [Results] The optimum pet dog dyeing process of plant hair dye was dye concentration of50 mg/mL,dye pH of 7. 5 and dyeing time of 1 h. [Conclusions] The coloring and fixing effects of the dye and dyeing process are good,which can lay a foundation for the development and utilization of plant dyes.
基金Peking University Comprehensive Platform for Innovative Drug Research and Development(Grant No.2009ZX--09301--010)
文摘We have developed an on-line detection method using acridine orange as the fluorescence probe and applied this method to rapidly identify active compounds in herbal medicines. This on-line method was equipped with a high-performance liquid chromatography tandem diode array detector, electrospray ionization-ion-trap time-of-flight mass spectrometry and DNA- acridine orange fluorescence detection （HPLC-DAD-MSn-DNA-AO-FLD）. A large amount of information could be simultaneously obtained during one run, which included HPLC fingerprint, ultraviolet spectra, total ion chromatograms, MSn data of high-resolution mass spectrometry and activity profile of each compound binding with DNA. The method also provided information on structureactivity relationships and mechanism of interaction. We used this on-line method to identify five DNA-binding activity components from Lithospermum erythrorhizon sample for the first time. The result showed that the parent nucleus of shikonin derivatives could bind with DNA. The structure-activity relationship showed that the parent nucleus of shikonin derivatives plays a major role in DNA binding, not the carboxyl group on the side chain. This simple, rapid, high precision and good stability on-line method should be useful for compound separation, structural identification and screening of DNA-binding compounds in herbal medicines.
文摘目的:探讨紫草多糖是否通过诱导自噬而抑制H_(22)肝癌实体瘤细胞的增殖。方法:将40只KM小鼠随机分为模型组、环磷酰胺组(阳性药物,30 mg/kg,ip,每4 d给药1次)和紫草多糖低、高剂量组(100、300 mg/kg,ig,每天给药1次),每组10只。各组小鼠均腋下接种H_(22)细胞株复制H_(22)肝癌实体瘤模型,造模同时各给药组小鼠给予相应药物,模型组小鼠ig等体积生理盐水。末次给药后,记录并测定小鼠体质量、瘤质量系数和胸腺、脾脏指数;采用实时荧光定量聚合酶链式反应法测定小鼠瘤组织中自噬相关基因Atg5、Beclin1 m RNA表达水平;Western blot法测定瘤组织中自噬微管相关蛋白1轻链3A/B(LC3A/B)蛋白表达水平。结果:紫草多糖可显著降低H_(22)实体瘤小鼠的瘤质量系数,抑瘤率达34.7%;可显著上调瘤组织中自噬相关基因Atg5、Beclin1 m RNA的表达和LC3A/B蛋白的表达,较模型组差异均有统计学意义(P<0.05或P<0.01)。结论:紫草多糖通过促进肝癌细胞的自噬,从而抑制H_(22)肝癌实体瘤细胞的增殖、延缓肿瘤生长。
文摘Crown gall tissue culture lines of Lithospermum erythrorhizon were established by transformation of sterile seedlings with Agrobacterium tumefaciens C58. Nopaline was inspected in the crown gall and showed that the transformation was successfull. L6 line, can secret red naphthoquinone pigments that are shikonin derivatives on phytohormone free 6,7-V medium, was selected from other lines on different basic phytohormone free media(MS, 6,7-V, WP, B5).The content of shikonin in the crown gall was 1 18%(dry tissue).The study showed that crown gall tissue culture of L.erythrorhizon was a new method to produce shikonin.
