Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to ident...Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to identifying potential therapeutic interventions. Cardamonin(CDN), the major active ingredient of the traditional Chinese medicinal herb Alpinia katsumadai, has exerted an excellent anti-inflammatory activity, but the mechanism underlying this role is not fully understood. Here, we show that CDN blocks canonical and noncanonical NLRP3 inflammasome activation triggered by multiple stimuli. Moreover, the suppression of CDN on inflammasome activation is specific to NLRP3, not to NLRC4 or AIM2 inflammasome. Besides, the inhibitory effect is not dependent on the expression of NF-κB-mediated inflammasome precursor proteins. We also demonstrate that CDN suppresses the NLRP3 inflammasome through blocking ASC oligomerization and speckle formation in a dose-dependent manner.Importantly, CDN improves the survival of mice suffering from lethal septic shock and attenuates IL-1βproduction induced by LPS in vivo, which is shown to be NLRP3 dependent. In conclusion, our results identify CDN as a broad-spectrum and specific inhibitor of NLRP3 inflammasome and a candidate therapeutic drug for treating NLRP3 inflammasome-driven diseases.展开更多
A rapid and accurate ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry(UPLC-MS/MS)method was established and validated for the measurement of two forms of cardamonin,i.e.,fr...A rapid and accurate ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry(UPLC-MS/MS)method was established and validated for the measurement of two forms of cardamonin,i.e.,free and glucuronidated,in the plasma and bile of rats.Cardamonin and an internal standard(1,8-dihydroxyanthraquinone)were extracted from plasma and bile with ethyl ether via liquid-liquid extraction.The analytes in the extracts were separated by an Agilent Zorbax Stable Bond-C18 column(2.1 mm×50 mm,1.8μm)under isocratic elution conditions[acetonitrile(A)and 0.1%ammonium formate in water(B),40:60(volume ratio)]with a flow rate of 0.4 mL/min,and mass spectrometry in negative ion MRM mode was implemented for analysis.Good linearity over the wide ranges of 0.01–5μg/mL for plasma and 0.025–10μg/mL for bile samples was acquired.Method validation was performed according to the FDA guidelines for bioanalytical methods.展开更多
目的探讨小豆蔻明(Cardamonin,CAR)抑制Raptor(Regulatory associated protein of mTOR)蛋白表达而调控HepG2细胞增殖和凋亡的作用。方法生物信息学分析TCGA(The Cancer Genome Atlas)数据库中肝癌患者RPTOR基因表达的差异及其与患者总...目的探讨小豆蔻明(Cardamonin,CAR)抑制Raptor(Regulatory associated protein of mTOR)蛋白表达而调控HepG2细胞增殖和凋亡的作用。方法生物信息学分析TCGA(The Cancer Genome Atlas)数据库中肝癌患者RPTOR基因表达的差异及其与患者总体生存率、无进展生存期等预后指标关系;CCK-8检测细胞活力;EdU核染色检测细胞增殖;流式细胞术检测细胞凋亡;Western Blot检测凋亡蛋白cleaved caspase 3、mTORC1信号通路关键蛋白mTOR、p-mTOR、S6K1、p-S6K1及mTORC1特异性结合蛋白Raptor的表达。结果RPTOR基因在肝癌组织中的表达水平高于正常组织(P<0.05),且高表达患者的总体生存率和无进展生存期显著低于低表达的患者(P<0.05);CAR抑制HepG2细胞增殖,诱导HepG2细胞凋亡,增加凋亡相关蛋白cleaved caspase3的表达;CAR降低肝癌HepG2细胞Raptor蛋白的表达,抑制mTOR、S6K1蛋白的磷酸化,但对其总蛋白的表达无明显影响。结论RPTOR基因在肝癌组织中高表达,与总体生存率和无进展生存期呈负相关;CAR可抑制肝癌HepG2细胞的增殖,促进其凋亡,机制与CAR降低Raptor的表达抑制mTORC1通路活性有关。展开更多
文摘目的观察豆蔻明(Cardamonin,CDN)对肺纤维化小鼠的作用,并探究其对转化生长因子-β1(transforming growth factor-β1,TGF-β1)/smad信号通路的影响。方法将小鼠随机分为生理盐水(Sham)组、博莱霉素(Bleomycin,BLM)组、豆蔻明低剂量(low dose of Cardamonin,CDN-L)组、豆蔻明中剂量(medium dose of Cardamonin,CDN-M)组、豆蔻明高剂量(high dose of Cardamonin,CDN-H)组、地塞米松(dexamethasone,DXM)组;单次气管注射BLM诱导肺纤维化,计算肺指数;酶联免疫吸附法测血清肿瘤坏死因子(tumor necrosis factor-α,TNF-α)水平;试剂盒测肺组织羟脯氨酸(hydroxyproline,HYP)含量,苏木精-伊红、马松染色观察肺炎症程度及纤维化情况,RT-PCR法检测肺组织TGF-β1/smad信号通路相关基因表达情况。结果与Sham组相比,BLM组小鼠肺指数、Szapiel评分、Ashcroft评分明显升高(P<0.05),肺部炎症程度和纤维化程度严重。血清中TNF-α含量升高(P<0.05)、肺组织HYP水平升高(P<0.05),TGF-β1[(1.02±0.21)vs.(3.25±0.14)]、smad2[(1.00±0.05)vs.(1.63±0.09)]、smad3[(1.00±0.06)vs.(1.82±0.07)]、α-平滑肌肌动蛋白(α-smoothmuscleactin,α-SMA)[(1.00±0.10)vs.(2.15±0.10)]mRNA表达升高(P<0.05),上皮-钙黏蛋白(E-Cadherin)[(1.01±0.16)vs.(0.57±0.09)]mRNA水平降低(P<0.05)。CDN-M、CDN-H干预可减低小鼠肺指数,减轻炎性和肺纤维化,降低TNF-α、HYP水平(P<0.05),同时下调肺纤维小鼠肺组织中TGF-β1、smad2、smad3、α-SMA表达,上调E-Cadherin表达。CDN-L无明显治疗效果,CDN-H治疗作用与DXM相似。结论CDN可能通过TGF-β1/Smad通路介导上皮-间充质转化而发挥抗纤维化作用。
基金supported by the National Key Technology R&D Program(No.2017ZX09301022)the National Key Technology R&D Program(No.2015ZX09501-004-001-008)+1 种基金the BeijingNova program(No.Z181100006218001)the National Natural Science Foundation of China(No.81630100)
文摘Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to identifying potential therapeutic interventions. Cardamonin(CDN), the major active ingredient of the traditional Chinese medicinal herb Alpinia katsumadai, has exerted an excellent anti-inflammatory activity, but the mechanism underlying this role is not fully understood. Here, we show that CDN blocks canonical and noncanonical NLRP3 inflammasome activation triggered by multiple stimuli. Moreover, the suppression of CDN on inflammasome activation is specific to NLRP3, not to NLRC4 or AIM2 inflammasome. Besides, the inhibitory effect is not dependent on the expression of NF-κB-mediated inflammasome precursor proteins. We also demonstrate that CDN suppresses the NLRP3 inflammasome through blocking ASC oligomerization and speckle formation in a dose-dependent manner.Importantly, CDN improves the survival of mice suffering from lethal septic shock and attenuates IL-1βproduction induced by LPS in vivo, which is shown to be NLRP3 dependent. In conclusion, our results identify CDN as a broad-spectrum and specific inhibitor of NLRP3 inflammasome and a candidate therapeutic drug for treating NLRP3 inflammasome-driven diseases.
文摘A rapid and accurate ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry(UPLC-MS/MS)method was established and validated for the measurement of two forms of cardamonin,i.e.,free and glucuronidated,in the plasma and bile of rats.Cardamonin and an internal standard(1,8-dihydroxyanthraquinone)were extracted from plasma and bile with ethyl ether via liquid-liquid extraction.The analytes in the extracts were separated by an Agilent Zorbax Stable Bond-C18 column(2.1 mm×50 mm,1.8μm)under isocratic elution conditions[acetonitrile(A)and 0.1%ammonium formate in water(B),40:60(volume ratio)]with a flow rate of 0.4 mL/min,and mass spectrometry in negative ion MRM mode was implemented for analysis.Good linearity over the wide ranges of 0.01–5μg/mL for plasma and 0.025–10μg/mL for bile samples was acquired.Method validation was performed according to the FDA guidelines for bioanalytical methods.
文摘目的探讨小豆蔻明(Cardamonin,CAR)抑制Raptor(Regulatory associated protein of mTOR)蛋白表达而调控HepG2细胞增殖和凋亡的作用。方法生物信息学分析TCGA(The Cancer Genome Atlas)数据库中肝癌患者RPTOR基因表达的差异及其与患者总体生存率、无进展生存期等预后指标关系;CCK-8检测细胞活力;EdU核染色检测细胞增殖;流式细胞术检测细胞凋亡;Western Blot检测凋亡蛋白cleaved caspase 3、mTORC1信号通路关键蛋白mTOR、p-mTOR、S6K1、p-S6K1及mTORC1特异性结合蛋白Raptor的表达。结果RPTOR基因在肝癌组织中的表达水平高于正常组织(P<0.05),且高表达患者的总体生存率和无进展生存期显著低于低表达的患者(P<0.05);CAR抑制HepG2细胞增殖,诱导HepG2细胞凋亡,增加凋亡相关蛋白cleaved caspase3的表达;CAR降低肝癌HepG2细胞Raptor蛋白的表达,抑制mTOR、S6K1蛋白的磷酸化,但对其总蛋白的表达无明显影响。结论RPTOR基因在肝癌组织中高表达,与总体生存率和无进展生存期呈负相关;CAR可抑制肝癌HepG2细胞的增殖,促进其凋亡,机制与CAR降低Raptor的表达抑制mTORC1通路活性有关。
