Curcuminoid difluoroboron has attractive performance as a promising near-infrared（NIR） fluorescent dye. In this contribution, we designed and synthesized a mitochondria-targeted NIR fluorescent probe DFB1 based on d...Curcuminoid difluoroboron has attractive performance as a promising near-infrared（NIR） fluorescent dye. In this contribution, we designed and synthesized a mitochondria-targeted NIR fluorescent probe DFB1 based on difluoroboron curcuminoid scaffold for the detection of Cys（cysteine）. DFB1 employs a curcumin analog as the NIR fluorophore, an acrylate group containing a, b-unsaturated ketone as a functional trigger moiety for Cys, and a triphenylphosphonium（TPP） cation moiety for specifically targeting mitochondria. The remarkable shift of DFB1 with Cys was observed from 470 nm to 590 nm in absorption spectra and from 560 nm to 680 nm in emission spectra. Notably, DFB1 manifests significantly dual-channel and turn-on NIR fluorescent signals simultaneously in response to Cys concentration,which make it favorable for monitoring endogenous Cys activity in vivo. This probe has high sensitivity and selectivity for the detection of Cys over homocysteine（Hcy） and glutathione（GSH）. This specific response for Cys was based on differences kinetics of intramolecular adduct/cyclizations. More importantly, biological experiments indicated that this probe could be utilized for the detection of endogenous mitochondrial Cys in living cells.展开更多
Objective:The present study aimed to evaluate the effect of a high water-soluble curcuminoids-rich extract(CRE)in a solid dispersion form(CRE-SD)using polyvinylpyrrolidone K30 on osteogenic induction of MC3T3-E1 cells...Objective:The present study aimed to evaluate the effect of a high water-soluble curcuminoids-rich extract(CRE)in a solid dispersion form(CRE-SD)using polyvinylpyrrolidone K30 on osteogenic induction of MC3T3-E1 cells.Methods:CRE was pre-purified using a microwave assisted extraction couple with a Diaion;HP-20 column chromatography.The osteoblastic cell proliferation and differentiation potentials of CRE-SD in MC3T3-E1 cells were tested by cell viability,alkaline phosphatase(ALP)activity,and Alizarin red S activity assays.The m RNA expressions of osteoblast-specific genes and underline mechanisms were assessed by a real time PCR and western blot analysis.Results:CRE-SD 50μg/m L increased alkaline phosphatase(ALP)activity,an early differentiation marker of osteoblasts in both MC3T3-E1 cells and non-osteogenic mouse pluripotent cell line,C3H10T1/2,indicating the action of CRE-SD was not cell-type specific.Alizarin red S activity showed a significant amount of calcium deposition in cells treated with CRE-SD.CRE-SD also upregulated the m RNA expression levels of transcription factors that favor osteoblast differentiation including Bmp-2,Runx2 and Collagen 1 a,in a dose dependent manner.Western blot analysis revealed that noggin attenuated CRE-SD-promoted expressions of Bmp-2 and Runx2 proteins.si RNA mediated blocking of Wnt/β-catenin signaling pathway also annulled the influence of CRE-SD,indicating Wnt/β-catenin dependent activity.Inhibition of the different signaling pathways abolished the influence of CRE-SD on ALP activity,confirming that CRE-SD induced MC3T3-E1 cells into osteoblasts through Wnt/β-catenin and BMP signaling pathway.Conclusion:These results collectively demonstrate that CRE-SD may be a potential therapeutic agent for the treatment of osteoporosis.展开更多
Leaf and tuber extracts of Curcuma wenyujin contain a mixture of curcuminoids.However,the curcuminoid constituents and their molecular mechanisms are poorly understood,and the relevant curcumin synthases remain unclea...Leaf and tuber extracts of Curcuma wenyujin contain a mixture of curcuminoids.However,the curcuminoid constituents and their molecular mechanisms are poorly understood,and the relevant curcumin synthases remain unclear.In this study,we comprehensively compared the metabolite profiles of the leaf and tuber tissues of C.wenyujin.A total of 11 curcuminoid metabolites were identified and exhibited differentially changed contents in the leaf and tuber tissues.An integrated analysis of metabolomic and transcriptomic data revealed the proposed biosynthesis pathway of curcuminoid.Two candidate type III polyketide synthases(PKSs)were identified in the metabolically engineering yeasts,indicating that CwPKS1 and CwPKS2 maintained substrate and product specificities.Especially,CwPKS1 is the first type III PKS identified to synthesize hydrogenated derivatives of curcuminoid,dihydrocurcumin and tetrehydrocurcumin.Interestingly,the substitution of the glycine at position 219 with aspartic acid(G219D mutant)resulted in the complete inactivation of CwPKS1.Our results provide the first comparative metabolome analysis of C.wenyujin and functionally identified type III PKSs,giving valuable information for curcuminoids biosynthesis.展开更多
基金supported by the National Natural Science Foundation of China for Excellent Young Scholars (No. 21622602)Distinguished Young Scholars (No. 21325625)+2 种基金 Oriental Scholarship, Fok Ying Tong Education Foundation (No. 142014)the Fundamental Research Funds for the Central Universities (Nos. WJ1616008, WK1013002)the State Key Laboratory of Fine Chemicals (No. KF1509)
文摘Curcuminoid difluoroboron has attractive performance as a promising near-infrared（NIR） fluorescent dye. In this contribution, we designed and synthesized a mitochondria-targeted NIR fluorescent probe DFB1 based on difluoroboron curcuminoid scaffold for the detection of Cys（cysteine）. DFB1 employs a curcumin analog as the NIR fluorophore, an acrylate group containing a, b-unsaturated ketone as a functional trigger moiety for Cys, and a triphenylphosphonium（TPP） cation moiety for specifically targeting mitochondria. The remarkable shift of DFB1 with Cys was observed from 470 nm to 590 nm in absorption spectra and from 560 nm to 680 nm in emission spectra. Notably, DFB1 manifests significantly dual-channel and turn-on NIR fluorescent signals simultaneously in response to Cys concentration,which make it favorable for monitoring endogenous Cys activity in vivo. This probe has high sensitivity and selectivity for the detection of Cys over homocysteine（Hcy） and glutathione（GSH）. This specific response for Cys was based on differences kinetics of intramolecular adduct/cyclizations. More importantly, biological experiments indicated that this probe could be utilized for the detection of endogenous mitochondrial Cys in living cells.
基金supported by the Thailand Research Fund (grant number RDG6150075)Prince of Songkla University (grant numbers MET611036S and MET6202025S)
文摘Objective:The present study aimed to evaluate the effect of a high water-soluble curcuminoids-rich extract(CRE)in a solid dispersion form(CRE-SD)using polyvinylpyrrolidone K30 on osteogenic induction of MC3T3-E1 cells.Methods:CRE was pre-purified using a microwave assisted extraction couple with a Diaion;HP-20 column chromatography.The osteoblastic cell proliferation and differentiation potentials of CRE-SD in MC3T3-E1 cells were tested by cell viability,alkaline phosphatase(ALP)activity,and Alizarin red S activity assays.The m RNA expressions of osteoblast-specific genes and underline mechanisms were assessed by a real time PCR and western blot analysis.Results:CRE-SD 50μg/m L increased alkaline phosphatase(ALP)activity,an early differentiation marker of osteoblasts in both MC3T3-E1 cells and non-osteogenic mouse pluripotent cell line,C3H10T1/2,indicating the action of CRE-SD was not cell-type specific.Alizarin red S activity showed a significant amount of calcium deposition in cells treated with CRE-SD.CRE-SD also upregulated the m RNA expression levels of transcription factors that favor osteoblast differentiation including Bmp-2,Runx2 and Collagen 1 a,in a dose dependent manner.Western blot analysis revealed that noggin attenuated CRE-SD-promoted expressions of Bmp-2 and Runx2 proteins.si RNA mediated blocking of Wnt/β-catenin signaling pathway also annulled the influence of CRE-SD,indicating Wnt/β-catenin dependent activity.Inhibition of the different signaling pathways abolished the influence of CRE-SD on ALP activity,confirming that CRE-SD induced MC3T3-E1 cells into osteoblasts through Wnt/β-catenin and BMP signaling pathway.Conclusion:These results collectively demonstrate that CRE-SD may be a potential therapeutic agent for the treatment of osteoporosis.
基金supported by the National Natural Science Foundation of China(82173919,82104320)the Natural Science Foundation of Zhejiang Province(LY21C050004,LQ22H280013).
文摘Leaf and tuber extracts of Curcuma wenyujin contain a mixture of curcuminoids.However,the curcuminoid constituents and their molecular mechanisms are poorly understood,and the relevant curcumin synthases remain unclear.In this study,we comprehensively compared the metabolite profiles of the leaf and tuber tissues of C.wenyujin.A total of 11 curcuminoid metabolites were identified and exhibited differentially changed contents in the leaf and tuber tissues.An integrated analysis of metabolomic and transcriptomic data revealed the proposed biosynthesis pathway of curcuminoid.Two candidate type III polyketide synthases(PKSs)were identified in the metabolically engineering yeasts,indicating that CwPKS1 and CwPKS2 maintained substrate and product specificities.Especially,CwPKS1 is the first type III PKS identified to synthesize hydrogenated derivatives of curcuminoid,dihydrocurcumin and tetrehydrocurcumin.Interestingly,the substitution of the glycine at position 219 with aspartic acid(G219D mutant)resulted in the complete inactivation of CwPKS1.Our results provide the first comparative metabolome analysis of C.wenyujin and functionally identified type III PKSs,giving valuable information for curcuminoids biosynthesis.
