4-Hydroxyphenylpyruvic acid （4-HPPA）, a kind of α-keto acid, is an intermediate in the metabolism of tyrosine and has a wide range of application in food, pharmaceutical and chemical industry. Using amino acids as ...4-Hydroxyphenylpyruvic acid （4-HPPA）, a kind of α-keto acid, is an intermediate in the metabolism of tyrosine and has a wide range of application in food, pharmaceutical and chemical industry. Using amino acids as raw material to prod uce the corresponding α-keto acid is thought to be both economic and efficient. Among the enzymes that convert amino acid to α-keto acid, membrane bound L-amino acid deaminase （mL-AAD）, which is anchored to the outer side of the cytomembrane, becomes an ideal enzyme to prepare α-keto acid since there is no cofactors needed and H2O2 production during the reaction. In this study, the mL-AAD from Proteus vulgaris was used to prepare whole-cell catalysts to produce 4-HPPA from L-tyrosine. The secretory efficiency of mL-AAD conducted by its own twin-arginine signal peptide （twin-arginine translocation pathway, Tat） and integrated pelB （the general secretory pathway, Sec）-Tat signal peptide was determined and compared firstly, using two pET systems （pET28a and pET20b）. It was found that the Tat pathway （pET28a-mlaad） resulted in higher cell-associated mL-AAD activity and cell biomass, and was more beneficial to prepare biocatalyst. In addition, expression hosts BI21 （DE3） and 0.05 mmol. L- 1 IPTG were found to be suitable for mL-AAD expression. The reaction conditions for mL-AAD were optimized and 72.72 mmol,L 1 4-HPPA was obtained from 100 mmol.L 1 tyrosine in 10 h under the optimized conditions. This bioprocess, which is more eco-friendly and economical than the traditional chemical synthesis ways, has great potential for industrial application.展开更多
Background:Recurrent miscarriage(RM)affects an estimated 1-3%of couples attempting to conceive,and its molecular components stay ineffectively caught on.This study aims to explore potential therapeutic targets for RM ...Background:Recurrent miscarriage(RM)affects an estimated 1-3%of couples attempting to conceive,and its molecular components stay ineffectively caught on.This study aims to explore potential therapeutic targets for RM by examining gene expression patterns and biological pathways in both mouse and human RM models.Meanwhile,explore relevant traditional Chinese medicine(TCM)components targeting potential therapeutic targets.Methods:We utilized the GSE211251 mouse and the GSE26787 human datasets,employing gene set enrichment analysis and gene metaphysics analysis to examine differentially expressed genes and enriched pathways.Single-cell RNA analysis uncovered cellular heterogeneity and arranged pharmacology-mapped potential drug-target intelligence.We employed molecular docking strategies to assess the affinity of TCM components for key proteins.Results:In the mouse model,genes such as Ly6f1 and Gpr26 were upregulated,while Stc5a and Galca exhibited downregulation.Gene set enrichment analysis identified key pathways,including the tumor necrosis factor-mediated signaling pathway.In human samples,Gene Ontology analysis highlighted processes such as apoptosis and cell adhesion.Single-cell RNA analysis revealed distinct cellular populations between normal and RM samples.Systems pharmacology identified C-X-C motif chemokine receptor 4(CXCR4)and endothelin 1(EDN1)as potential key targets,and molecular docking confirmed that stearic acid from TCM appears to regulate these proteins.Conclusion:This study presents a comprehensive analysis of the genetic and cellular underpinnings of RM,identifying CXCR4 and EDN1 as promising therapeutic targets.Stearic acid from TCM could provide targeted treatment by modulating these key proteins,paving the way for new RM treatment strategies.展开更多
This study aimed to evaluate the effects of Bifi dobacterium breve CCFM683 on psoriasis and to investigate the underlying mechanisms.B.breve CCFM683 significantly ameliorated psoriasis in mice as well as elevated the ...This study aimed to evaluate the effects of Bifi dobacterium breve CCFM683 on psoriasis and to investigate the underlying mechanisms.B.breve CCFM683 significantly ameliorated psoriasis in mice as well as elevated the deoxycholic acid(DCA)and lithocholic acid(LCA)in the colon compared with those of the imiquimod(IMQ)-treated mice.Meanwhile,B.breve CCFM683 increased the relative abundance of DCA-producing Lachnoclostridium and diminished the harmful Desulfovibrio and Prevotellaceae UCG001.Additionally,the farnesoid X receptor(FXR)in the skin was activated and the expression of the Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NF-κB)pathway was inhibited,and the downstream interleukin(IL)-17 and tumor necrosis factor(TNF)-αwere downregulated whereas IL-10 was up-regulated.Moreover,the subsequent hyperproliferation of keratinocytes and the dysfunction of the epidermal barrier were improved.In conclusion,CCFM683 administration ameliorated IMQ-induced psoriasis via modulating gut microbiota,promoting the DCA production,regulating the FXR-TLR4/NF-κB pathway,diminishing proinflammatory cytokines,and regulating keratinocytes and epidermal barrier.These findings may be conducive to elucidating the mechanism for probiotics to ameliorate psoriasis and to promote its clinical trials in skin disease.展开更多
[Objectives]To explore the effects of chlorogenic acid from honeysuckle on the secretion enzymes,lipoxygenase A4(LXA4),and blood biochemical indicators in mice with aluminum induced Alzheimer's disease(AD).[Method...[Objectives]To explore the effects of chlorogenic acid from honeysuckle on the secretion enzymes,lipoxygenase A4(LXA4),and blood biochemical indicators in mice with aluminum induced Alzheimer's disease(AD).[Methods]Chlorogenic acid was extracted from hon-eysuckle by ultrasound assisted alcohol extraction method.Seventy mice were randomly divided into normal group,model group,and low,me-dium and high dose groups of honeysuckle chlorogenic acid.All the mice in each group except for the normal group were given maltol aluminum by intraperitoneal injection to establish models of aluminum induced AD,continuously injected for 5 d and stopped for 2 d,totally poisoned for 8 weeks.Starting from the 5^(th) week of poisoning,the low,medium and high dose groups of honeysuckle chlorogenic acid were given honeysuck-le chlorogenc acid solution 40,80 and 160 mg/kg by gavage,respectively,while the normal group and the model group were fed with an equal volume of distilled water,all once daily,continuously gavaged until the end of the 8^(th) week.At the end of the experiment,the learning memory ability of the mice was tested by Y-type waler maze,and the number of tests required to reach the learning standard,the number of memory er-rors in 20 tests and the error rate of the mice were recorded.The brains of mice were taken to determine the contents of β-secretase,α-secre-tase,γ-secretase,LXA4 and acetylcholinesterase(AchE)in the homogenates of brain tissues by ELISA,and their blood was taken to deter-mine the biochemical indexes.[Results]Compared with the normal group,the number of learning tests,number of memory errors,error rate and the contents of β-secretase,γ-secretase and AchE in brain tissue of the mice in the model group were all significantly increased(all P<0.05),the contents of LXA4 in brain tissue were significantly decreased(all P<0.05),and the contents of α-secretase did not change significantly(all P>0.05);compared with the model group,the number of learning tests,the number of memory errors,the error rate and the content of β-secretase,γ-secretase and AchE in brain tissue were all significantly reduced(all P<0.05),the content of LXA4 in brain tissue of the high dose group of honeysuckle chlorogenic acid was significantly increased(P<0.05),and there was no significant change in the content of α-secretase in brain tissue of all groups of honeysuckle chlorogenic acid(all P>0.05).Compared with the normal group,the levels of blood glucose,TC,TG,ALT,BUN,Cr and UA in the model group and the levels of TC,TG and BUN in the low-and medium-dose groups of honeysuckle chlorogenic acid were significantly increased(all P<0.05),and the level of HDL-C in the model group and the levels of UA in the medium-and high-dose groups of honeysuckle chlorogenic acid were significantly decreased(all P<0.05);compared with the model group,the levels of blood glucose,ALT,BUN,UA in each group of honeysuckle chlorogenic acid,the levels of TC and Cr in medium and high dose groups of honeysuckle chlorogenic acid,and the level of TG in the high dose group of honeysuckle chlorogenic acid were all signifi-cantly lower(all P<0.05),while the level of HDL-C in the medium and high dose groups of honeysuckle chlorogenic acid and the level of to-tal protein in the high dose group of honeysuckle chlorogenic acid were all significantly higher(all P<0.05).[Conclusions]Chlorogenic acid from honeysuckle may improve AD induced by aluminum exposure via regulating related secretory enzymes,LXA4,and various biochemi-cal indicators.展开更多
Branched-chain fatty acids(BCFAs)are new bioactive fatty acids with anti-inflammatory properties.However,the role of BCFAs in alleviating ulcerative colitis has not been clarified.Herein,we evaluated the protective ef...Branched-chain fatty acids(BCFAs)are new bioactive fatty acids with anti-inflammatory properties.However,the role of BCFAs in alleviating ulcerative colitis has not been clarified.Herein,we evaluated the protective effect of BCFAs from goat milk in mice with colitis induced using dextran sodium sulfate(DSS)and explored the corresponding mechanism.These results show that BCFAs extracted from goat milk can significantly alleviate weight loss in mice,and reduce the disease activity index and the activity of myeloperoxidase while increasing the content of antioxidant enzymes in colon tissue and reducing the oxidation stress response.These data also show that BCFAs can down-regulate the gene and protein expression of the toll-like receptor 4(TLR4)/nuclear factorκB p65(NF-κB p65)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway,and at the same time significantly reduce the expression of pro-inflammatory factors tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),and IL-18 in colon tissue,and significantly increase the expression of the anti-inflammatory factor IL-10.In conclusion,these results demonstrated that BCFAs in goat milk exerted effects on colitis-related inflammatory cytokines and inhibited inflammation by inducing the TLR4/NF-κB/NLRP3 pathway to alleviate DSS-induced ulcerative colitis.This study provides evidence for the potential of BCFAs as bioactive fatty acids in food products and to ameliorate ulcerative colitis development in mice.展开更多
基金Supported by the National Natural Science Foundation of China(31470793,31670804)China Postdoctoral Science Foundation(2016M592003)+1 种基金the Natural Science Foundation of Zhejiang Province(LZ13B060002)the General Scientific Research Project of Zhejiang Provincial Education Department(Y201432760)
文摘4-Hydroxyphenylpyruvic acid （4-HPPA）, a kind of α-keto acid, is an intermediate in the metabolism of tyrosine and has a wide range of application in food, pharmaceutical and chemical industry. Using amino acids as raw material to prod uce the corresponding α-keto acid is thought to be both economic and efficient. Among the enzymes that convert amino acid to α-keto acid, membrane bound L-amino acid deaminase （mL-AAD）, which is anchored to the outer side of the cytomembrane, becomes an ideal enzyme to prepare α-keto acid since there is no cofactors needed and H2O2 production during the reaction. In this study, the mL-AAD from Proteus vulgaris was used to prepare whole-cell catalysts to produce 4-HPPA from L-tyrosine. The secretory efficiency of mL-AAD conducted by its own twin-arginine signal peptide （twin-arginine translocation pathway, Tat） and integrated pelB （the general secretory pathway, Sec）-Tat signal peptide was determined and compared firstly, using two pET systems （pET28a and pET20b）. It was found that the Tat pathway （pET28a-mlaad） resulted in higher cell-associated mL-AAD activity and cell biomass, and was more beneficial to prepare biocatalyst. In addition, expression hosts BI21 （DE3） and 0.05 mmol. L- 1 IPTG were found to be suitable for mL-AAD expression. The reaction conditions for mL-AAD were optimized and 72.72 mmol,L 1 4-HPPA was obtained from 100 mmol.L 1 tyrosine in 10 h under the optimized conditions. This bioprocess, which is more eco-friendly and economical than the traditional chemical synthesis ways, has great potential for industrial application.
基金support from the Ningxia Hui Autonomous Region Key Research and Development Program(Project No.2021BEG03041).
文摘Background:Recurrent miscarriage(RM)affects an estimated 1-3%of couples attempting to conceive,and its molecular components stay ineffectively caught on.This study aims to explore potential therapeutic targets for RM by examining gene expression patterns and biological pathways in both mouse and human RM models.Meanwhile,explore relevant traditional Chinese medicine(TCM)components targeting potential therapeutic targets.Methods:We utilized the GSE211251 mouse and the GSE26787 human datasets,employing gene set enrichment analysis and gene metaphysics analysis to examine differentially expressed genes and enriched pathways.Single-cell RNA analysis uncovered cellular heterogeneity and arranged pharmacology-mapped potential drug-target intelligence.We employed molecular docking strategies to assess the affinity of TCM components for key proteins.Results:In the mouse model,genes such as Ly6f1 and Gpr26 were upregulated,while Stc5a and Galca exhibited downregulation.Gene set enrichment analysis identified key pathways,including the tumor necrosis factor-mediated signaling pathway.In human samples,Gene Ontology analysis highlighted processes such as apoptosis and cell adhesion.Single-cell RNA analysis revealed distinct cellular populations between normal and RM samples.Systems pharmacology identified C-X-C motif chemokine receptor 4(CXCR4)and endothelin 1(EDN1)as potential key targets,and molecular docking confirmed that stearic acid from TCM appears to regulate these proteins.Conclusion:This study presents a comprehensive analysis of the genetic and cellular underpinnings of RM,identifying CXCR4 and EDN1 as promising therapeutic targets.Stearic acid from TCM could provide targeted treatment by modulating these key proteins,paving the way for new RM treatment strategies.
基金supported by the National Natural Science Foundation of China(32072227,32021005)111 Project(BP0719028)the Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province.
文摘This study aimed to evaluate the effects of Bifi dobacterium breve CCFM683 on psoriasis and to investigate the underlying mechanisms.B.breve CCFM683 significantly ameliorated psoriasis in mice as well as elevated the deoxycholic acid(DCA)and lithocholic acid(LCA)in the colon compared with those of the imiquimod(IMQ)-treated mice.Meanwhile,B.breve CCFM683 increased the relative abundance of DCA-producing Lachnoclostridium and diminished the harmful Desulfovibrio and Prevotellaceae UCG001.Additionally,the farnesoid X receptor(FXR)in the skin was activated and the expression of the Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NF-κB)pathway was inhibited,and the downstream interleukin(IL)-17 and tumor necrosis factor(TNF)-αwere downregulated whereas IL-10 was up-regulated.Moreover,the subsequent hyperproliferation of keratinocytes and the dysfunction of the epidermal barrier were improved.In conclusion,CCFM683 administration ameliorated IMQ-induced psoriasis via modulating gut microbiota,promoting the DCA production,regulating the FXR-TLR4/NF-κB pathway,diminishing proinflammatory cytokines,and regulating keratinocytes and epidermal barrier.These findings may be conducive to elucidating the mechanism for probiotics to ameliorate psoriasis and to promote its clinical trials in skin disease.
基金Supported by Baise Science Research and Technology Development Plan Project(20232022)Cuangxi College Students’Innovation and Entrepreneurship Training Program(Recommend National Level2022210599040S).
文摘[Objectives]To explore the effects of chlorogenic acid from honeysuckle on the secretion enzymes,lipoxygenase A4(LXA4),and blood biochemical indicators in mice with aluminum induced Alzheimer's disease(AD).[Methods]Chlorogenic acid was extracted from hon-eysuckle by ultrasound assisted alcohol extraction method.Seventy mice were randomly divided into normal group,model group,and low,me-dium and high dose groups of honeysuckle chlorogenic acid.All the mice in each group except for the normal group were given maltol aluminum by intraperitoneal injection to establish models of aluminum induced AD,continuously injected for 5 d and stopped for 2 d,totally poisoned for 8 weeks.Starting from the 5^(th) week of poisoning,the low,medium and high dose groups of honeysuckle chlorogenic acid were given honeysuck-le chlorogenc acid solution 40,80 and 160 mg/kg by gavage,respectively,while the normal group and the model group were fed with an equal volume of distilled water,all once daily,continuously gavaged until the end of the 8^(th) week.At the end of the experiment,the learning memory ability of the mice was tested by Y-type waler maze,and the number of tests required to reach the learning standard,the number of memory er-rors in 20 tests and the error rate of the mice were recorded.The brains of mice were taken to determine the contents of β-secretase,α-secre-tase,γ-secretase,LXA4 and acetylcholinesterase(AchE)in the homogenates of brain tissues by ELISA,and their blood was taken to deter-mine the biochemical indexes.[Results]Compared with the normal group,the number of learning tests,number of memory errors,error rate and the contents of β-secretase,γ-secretase and AchE in brain tissue of the mice in the model group were all significantly increased(all P<0.05),the contents of LXA4 in brain tissue were significantly decreased(all P<0.05),and the contents of α-secretase did not change significantly(all P>0.05);compared with the model group,the number of learning tests,the number of memory errors,the error rate and the content of β-secretase,γ-secretase and AchE in brain tissue were all significantly reduced(all P<0.05),the content of LXA4 in brain tissue of the high dose group of honeysuckle chlorogenic acid was significantly increased(P<0.05),and there was no significant change in the content of α-secretase in brain tissue of all groups of honeysuckle chlorogenic acid(all P>0.05).Compared with the normal group,the levels of blood glucose,TC,TG,ALT,BUN,Cr and UA in the model group and the levels of TC,TG and BUN in the low-and medium-dose groups of honeysuckle chlorogenic acid were significantly increased(all P<0.05),and the level of HDL-C in the model group and the levels of UA in the medium-and high-dose groups of honeysuckle chlorogenic acid were significantly decreased(all P<0.05);compared with the model group,the levels of blood glucose,ALT,BUN,UA in each group of honeysuckle chlorogenic acid,the levels of TC and Cr in medium and high dose groups of honeysuckle chlorogenic acid,and the level of TG in the high dose group of honeysuckle chlorogenic acid were all signifi-cantly lower(all P<0.05),while the level of HDL-C in the medium and high dose groups of honeysuckle chlorogenic acid and the level of to-tal protein in the high dose group of honeysuckle chlorogenic acid were all significantly higher(all P<0.05).[Conclusions]Chlorogenic acid from honeysuckle may improve AD induced by aluminum exposure via regulating related secretory enzymes,LXA4,and various biochemi-cal indicators.
基金financially supported by the 2021 Kabrita Nutrition Grant.
文摘Branched-chain fatty acids(BCFAs)are new bioactive fatty acids with anti-inflammatory properties.However,the role of BCFAs in alleviating ulcerative colitis has not been clarified.Herein,we evaluated the protective effect of BCFAs from goat milk in mice with colitis induced using dextran sodium sulfate(DSS)and explored the corresponding mechanism.These results show that BCFAs extracted from goat milk can significantly alleviate weight loss in mice,and reduce the disease activity index and the activity of myeloperoxidase while increasing the content of antioxidant enzymes in colon tissue and reducing the oxidation stress response.These data also show that BCFAs can down-regulate the gene and protein expression of the toll-like receptor 4(TLR4)/nuclear factorκB p65(NF-κB p65)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway,and at the same time significantly reduce the expression of pro-inflammatory factors tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),and IL-18 in colon tissue,and significantly increase the expression of the anti-inflammatory factor IL-10.In conclusion,these results demonstrated that BCFAs in goat milk exerted effects on colitis-related inflammatory cytokines and inhibited inflammation by inducing the TLR4/NF-κB/NLRP3 pathway to alleviate DSS-induced ulcerative colitis.This study provides evidence for the potential of BCFAs as bioactive fatty acids in food products and to ameliorate ulcerative colitis development in mice.
