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
Objective:To investigate the potential anti-tumor mechanisms of naphthoquinone compound shikonin(SKN)extracted from the root of Chinese herbal medicine plant lithospermum(Lithospermum erythrorhizon Sieb.&Zucc.).Me...Objective:To investigate the potential anti-tumor mechanisms of naphthoquinone compound shikonin(SKN)extracted from the root of Chinese herbal medicine plant lithospermum(Lithospermum erythrorhizon Sieb.&Zucc.).Methods:We first observed that SKN treatment led to swelling and bubbles in HeLa cells that were similar to the phenotype of cell pyroptosis.Subsequently,the HeLa cells experienced a pyroptotic process with SKN,and this was then assessed using lactate dehydrogenase(LDH)release and propidium iodide(PI)/Hoechst double staining experiments.Pyroptosis is defined as gasdermin-mediated programmed necroptosis.To identify the potential pyroptosis machinery,two strategies were utilized that included a genome-wide clustered regularly interspaced short palindromic repeats(CRISPR)-associated protein 9 screening experiment and a pyroptosis reconstitution assay executed by each of the five known gasdermins(GSDMA-E).Moreover,endogenous cleavage was also detected in a panel of tumor cell lines.Results:Compared with the control,both the LDH release and PI/Hoechst double-staining experiments suggested that SKN induced perforation and enhancement of the permeability of the cell membranes that resulted in pyroptosis in HeLa cells(P=.028 and P=.032,respectively).In addition,the reconstitution assays in human embryonic kidney 293T(HEK-293T)cells and endogenous cleavage assays in HeLa cells indicated that the pyroptosis was controlled by GSDME.In addition,we also found SKN could trigger pyroptosis in a panel of tumor cell lines in which the cellular morphologies were proportional to the GSDME expression levels.Additionally,the cleavage of GSDME was also detected,and this was indicative of a similar GSDME-mediated mechanism.Conclusion:Our study not only explained the molecular mechanism of cytotoxicity of SKN to various tumor cells,but also provided additional information for the potential clinical application of natural naphthoquinone compounds against cancer.展开更多
Pogostemon cablin essential oil(PEO),extracted from P.cablin,has anti-oxidant,anti-inflammatory,and anti-stress properties,as well as the ability to improve gastrointestinal digestion.This study aims to evaluate the e...Pogostemon cablin essential oil(PEO),extracted from P.cablin,has anti-oxidant,anti-inflammatory,and anti-stress properties,as well as the ability to improve gastrointestinal digestion.This study aims to evaluate the effects of PEO on the performance,rumen epithelial morphology,and barrier function in heat-stressed beef cattle.Thirty-six male Jingjiang cattle at 18 months old were randomly assigned into four groups and fed a diet containing PEO at 0(control),50,100,or 150 mg/kg in the feed concentrate(n=9).All experimental cattle were fed under high temperature and humidity in summer for 60 days.The results indicated that 50 mg/kg of PEO treatment enhanced the average daily gain of beef cattle compared with the control group(P=0.032).All PEO treatments reduced the diamine oxidase activity(P=0.004)and malondialdehyde content(P=0.008)in serum.In addition,the content of 70 kDa heat shock protein in the 100 mg/kg group was increased,and the activity of glutathione peroxidase and total antioxidant capacity in both 100 mg/kg and 150 mg/kg groups were enhanced compared to the control group(P<0.05).More importantly,PEO treatment with 50 mg/kg enhanced the mRNA relative expressions of occludin in ruminal epithelia but decreased the mRNA relative expressions of c-Jun N-terminal kinase,P38 mitogen-activated protein kinases,caspase-3,Beclin1(P<0.05),and extremely significant declined the mRNA relative expressions of extracellular regulated protein kinases and ubiquitin-binding protein in contrast to the control group(P<0.01).These findings indicated that dietary PEO supplementation might be favorable to improve growth performance and repairing damaged rumen epithelium of heat-stressed cattle by down-regulating the mitogen-activated protein kinase signaling pathway.展开更多
Via an insufficient coat protein complex I(COPI)retrieval signal,the majority of SARSCo V-2 spike(S)is resident in host early secretory organelles and a tiny amount is leaked out in cell surface.Only surface-exposed S...Via an insufficient coat protein complex I(COPI)retrieval signal,the majority of SARSCo V-2 spike(S)is resident in host early secretory organelles and a tiny amount is leaked out in cell surface.Only surface-exposed S can be recognized by B cell receptor(BCR)or anti-S therapeutic monoclonal antibodies(m Abs)that is the trigger step for B cell activation after S m RNA vaccination or infected cell clearance by S m Abs.Now,a drug strategy to promote S host surface exposure is absent.Here,we first combined structural and biochemical analysis to characterize S COPI sorting signals.A potent S COPI sorting inhibitor was then invented,evidently capable of promoting S surface exposure and facilitating infected cell clearance by S antibody-dependent cellular cytotoxicity(ADCC).Importantly,with the inhibitor as a probe,we revealed Omicron BA.1 S is less cell surface exposed than prototypes because of a constellation of S folding mutations,possibly corresponding to its ER chaperone association.Our findings not only suggest COPI is a druggable target against COVID-19,but also highlight SARS-Co V-2 evolution mechanism driven by S folding and trafficking mutations.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This research was supported by the Program for the Research Startup Fund Program at the Beijing University of Chinese Medicine(90011451310011)the Key Research Fund for Drug Discovery in Chinese Medicine at the Beijing University of Chinese Medicine(1000061223740)+1 种基金the Experimental Technology Standardization Research Project at the Beijing University of Chinese Medicine(2021-SYJS-009)the Fundamental Research Funds for the Central Universities of the Beijing University of Chinese Medicine(2020-JYB-ZDGG-057).
文摘Objective:To investigate the potential anti-tumor mechanisms of naphthoquinone compound shikonin(SKN)extracted from the root of Chinese herbal medicine plant lithospermum(Lithospermum erythrorhizon Sieb.&Zucc.).Methods:We first observed that SKN treatment led to swelling and bubbles in HeLa cells that were similar to the phenotype of cell pyroptosis.Subsequently,the HeLa cells experienced a pyroptotic process with SKN,and this was then assessed using lactate dehydrogenase(LDH)release and propidium iodide(PI)/Hoechst double staining experiments.Pyroptosis is defined as gasdermin-mediated programmed necroptosis.To identify the potential pyroptosis machinery,two strategies were utilized that included a genome-wide clustered regularly interspaced short palindromic repeats(CRISPR)-associated protein 9 screening experiment and a pyroptosis reconstitution assay executed by each of the five known gasdermins(GSDMA-E).Moreover,endogenous cleavage was also detected in a panel of tumor cell lines.Results:Compared with the control,both the LDH release and PI/Hoechst double-staining experiments suggested that SKN induced perforation and enhancement of the permeability of the cell membranes that resulted in pyroptosis in HeLa cells(P=.028 and P=.032,respectively).In addition,the reconstitution assays in human embryonic kidney 293T(HEK-293T)cells and endogenous cleavage assays in HeLa cells indicated that the pyroptosis was controlled by GSDME.In addition,we also found SKN could trigger pyroptosis in a panel of tumor cell lines in which the cellular morphologies were proportional to the GSDME expression levels.Additionally,the cleavage of GSDME was also detected,and this was indicative of a similar GSDME-mediated mechanism.Conclusion:Our study not only explained the molecular mechanism of cytotoxicity of SKN to various tumor cells,but also provided additional information for the potential clinical application of natural naphthoquinone compounds against cancer.
基金The National Natural Science Foundation of China (32060768)Central guidance for local scientifc and technological development funding projects of China (20231zdf03083)the China Agriculture Research System of MOF and MARA (CARS-37)provided financial support for this study.
文摘Pogostemon cablin essential oil(PEO),extracted from P.cablin,has anti-oxidant,anti-inflammatory,and anti-stress properties,as well as the ability to improve gastrointestinal digestion.This study aims to evaluate the effects of PEO on the performance,rumen epithelial morphology,and barrier function in heat-stressed beef cattle.Thirty-six male Jingjiang cattle at 18 months old were randomly assigned into four groups and fed a diet containing PEO at 0(control),50,100,or 150 mg/kg in the feed concentrate(n=9).All experimental cattle were fed under high temperature and humidity in summer for 60 days.The results indicated that 50 mg/kg of PEO treatment enhanced the average daily gain of beef cattle compared with the control group(P=0.032).All PEO treatments reduced the diamine oxidase activity(P=0.004)and malondialdehyde content(P=0.008)in serum.In addition,the content of 70 kDa heat shock protein in the 100 mg/kg group was increased,and the activity of glutathione peroxidase and total antioxidant capacity in both 100 mg/kg and 150 mg/kg groups were enhanced compared to the control group(P<0.05).More importantly,PEO treatment with 50 mg/kg enhanced the mRNA relative expressions of occludin in ruminal epithelia but decreased the mRNA relative expressions of c-Jun N-terminal kinase,P38 mitogen-activated protein kinases,caspase-3,Beclin1(P<0.05),and extremely significant declined the mRNA relative expressions of extracellular regulated protein kinases and ubiquitin-binding protein in contrast to the control group(P<0.01).These findings indicated that dietary PEO supplementation might be favorable to improve growth performance and repairing damaged rumen epithelium of heat-stressed cattle by down-regulating the mitogen-activated protein kinase signaling pathway.
基金supported by Startup fund program at Beijing University of Chinese Medicine(BUCM)(90011451310011,China)to Wenfu Mathe emergency fund against COVID-19 program at BUCM(1000061223476,China)to Wenfu Mathe innovation team and talents cultivation program of national administration of traditional Chinese medicine(ZYYCXTD-C202006,China)to Wenfu Ma。
文摘Via an insufficient coat protein complex I(COPI)retrieval signal,the majority of SARSCo V-2 spike(S)is resident in host early secretory organelles and a tiny amount is leaked out in cell surface.Only surface-exposed S can be recognized by B cell receptor(BCR)or anti-S therapeutic monoclonal antibodies(m Abs)that is the trigger step for B cell activation after S m RNA vaccination or infected cell clearance by S m Abs.Now,a drug strategy to promote S host surface exposure is absent.Here,we first combined structural and biochemical analysis to characterize S COPI sorting signals.A potent S COPI sorting inhibitor was then invented,evidently capable of promoting S surface exposure and facilitating infected cell clearance by S antibody-dependent cellular cytotoxicity(ADCC).Importantly,with the inhibitor as a probe,we revealed Omicron BA.1 S is less cell surface exposed than prototypes because of a constellation of S folding mutations,possibly corresponding to its ER chaperone association.Our findings not only suggest COPI is a druggable target against COVID-19,but also highlight SARS-Co V-2 evolution mechanism driven by S folding and trafficking mutations.
