A large amount of evidence has supported a clinical link between diabetes and inflammatory diseases,e.g.,cancer,dementia,and hypertension.In addition,it is also suggested that dysregulations related to Ca^(2+)signalin...A large amount of evidence has supported a clinical link between diabetes and inflammatory diseases,e.g.,cancer,dementia,and hypertension.In addition,it is also suggested that dysregulations related to Ca^(2+)signaling could link these diseases,in addition to 3'-5'-cyclic adenosine monophosphate(cAMP)signaling pathways.Thus,revealing this interplay between diabetes and inflammatory diseases may provide novel insights into the pathogenesis of these diseases.Publications involving signaling pathways related to Ca^(2+)and cAMP,inflammation,diabetes,dementia,cancer,and hypertension(alone or combined)were collected by searching PubMed and EMBASE.Both signaling pathways,Ca^(2+)and cAMP signaling,control the release of neurotransmitters and hormones,in addition to neurodegeneration,and tumor growth.Furthermore,there is a clear relationship between Ca^(2+)signaling,e.g.,increased Ca^(2+)signals,and inflammatory responses.cAMP also regulates pro-and anti-inflammatory responses.Due to the experience of our group in this field,this article discusses the role of Ca^(2+)and cAMP signaling in the correlation between diabetes and inflammatory diseases,including its pharmacological implications.As a novelty,this article also includes:(1)A timeline of the major events in Ca^(2+)/cAMP signaling;and(2)As coronavirus disease 2019(COVID-19)is an emerging and rapidly evolving situation,this article also discusses recent reports on the role of Ca^(2+)channel blockers for preventing Ca^(2+)signaling disruption due to COVID-19,including the correlation between COVID-19 and diabetes.展开更多
In this study,we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate(c AMP)from adenosine triphosphate(ATP).First,adenylate cyclase from Escherichia coli MG1655(E...In this study,we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate(c AMP)from adenosine triphosphate(ATP).First,adenylate cyclase from Escherichia coli MG1655(EAC)and Bordetella Pertussis(BAC)were expressed in E.coli BL21(DE3)and comparatively analyzed for their activities.As a result,EAC from E.coli MG1655 exhibited a higher activity.However,amount of EAC were obtained in an insoluble form.Therefore,we expressed the first 446 amino acids of EAC(EAC446)to avoid the inclusion body.The effects of induction temperature,incubation time,and incubation p H were further evaluated to improve the expression of EAC446.Subsequently,the reaction process for the production of c AMP with ATP as a starting material was investigated.As none of c AMP was detected in the whole-cell based biocatalytic process,the reaction catalyzed by the crude enzyme was determined for c AMP production.What's more,the reaction temperature,reaction p H,metal ion additives and substrate concentration was optimized,and the maximum c AMP production of 18.45 g·L^-1was achieved with a yield of 95.4%after bioconversion of 6 h.展开更多
The unicellular green alga,Chlamydomonas reinhardtii is a model organism for studying various biological processes,such as photosynthesis,flagellar motility,and lipid metabolism.To find some novel genes regulating the...The unicellular green alga,Chlamydomonas reinhardtii is a model organism for studying various biological processes,such as photosynthesis,flagellar motility,and lipid metabolism.To find some novel genes regulating the lipid metabolism under various stress conditions,the paromomycin resistance gene aphVIII was transferred into the genome of C.reinhardtii to establish a mutant library.Two genes mutated in two of the TAG-reduced mutants(Cre06.g278111 in M2 mutant,Cre06.g278110 in M6 mutants)were neighboring in the genome,and their expression levels were down-regulated in their corresponding mutants in parallel with their reduced TAG levels following N deprivation.The proteins encoded by these two genes(KCN11 by Cre06.g278111,ACYC3 by Cre06.g278110)contained a conversed cyclic mononucleotide phosphate(cNMP)binding protein and an adenylate domain,respectively.Since cNMP binding protein and adenylate domain have been known as important components of cyclic adenosine monophosphate(cAMP)signaling pathway,suggesting that these two genes might af fect cellular TAG biosynthesis through cAMP signal pathway.展开更多
To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role...To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role of hemin on this pathway, HTMCs of the third to fourth generation were cultured in vitro. Reverse transcripase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.展开更多
文摘A large amount of evidence has supported a clinical link between diabetes and inflammatory diseases,e.g.,cancer,dementia,and hypertension.In addition,it is also suggested that dysregulations related to Ca^(2+)signaling could link these diseases,in addition to 3'-5'-cyclic adenosine monophosphate(cAMP)signaling pathways.Thus,revealing this interplay between diabetes and inflammatory diseases may provide novel insights into the pathogenesis of these diseases.Publications involving signaling pathways related to Ca^(2+)and cAMP,inflammation,diabetes,dementia,cancer,and hypertension(alone or combined)were collected by searching PubMed and EMBASE.Both signaling pathways,Ca^(2+)and cAMP signaling,control the release of neurotransmitters and hormones,in addition to neurodegeneration,and tumor growth.Furthermore,there is a clear relationship between Ca^(2+)signaling,e.g.,increased Ca^(2+)signals,and inflammatory responses.cAMP also regulates pro-and anti-inflammatory responses.Due to the experience of our group in this field,this article discusses the role of Ca^(2+)and cAMP signaling in the correlation between diabetes and inflammatory diseases,including its pharmacological implications.As a novelty,this article also includes:(1)A timeline of the major events in Ca^(2+)/cAMP signaling;and(2)As coronavirus disease 2019(COVID-19)is an emerging and rapidly evolving situation,this article also discusses recent reports on the role of Ca^(2+)channel blockers for preventing Ca^(2+)signaling disruption due to COVID-19,including the correlation between COVID-19 and diabetes.
基金The National Natural Science Foundation of China(Grant No.21576134,Grant No.21606127,Grant No.21390200,Grant No.21706126)the National Key Research and Development Program of China(Grant No.2016YFA0204300)the Top-notch Academic Programs Project of Jiangsu Higher Education Institutions。
文摘In this study,we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate(c AMP)from adenosine triphosphate(ATP).First,adenylate cyclase from Escherichia coli MG1655(EAC)and Bordetella Pertussis(BAC)were expressed in E.coli BL21(DE3)and comparatively analyzed for their activities.As a result,EAC from E.coli MG1655 exhibited a higher activity.However,amount of EAC were obtained in an insoluble form.Therefore,we expressed the first 446 amino acids of EAC(EAC446)to avoid the inclusion body.The effects of induction temperature,incubation time,and incubation p H were further evaluated to improve the expression of EAC446.Subsequently,the reaction process for the production of c AMP with ATP as a starting material was investigated.As none of c AMP was detected in the whole-cell based biocatalytic process,the reaction catalyzed by the crude enzyme was determined for c AMP production.What's more,the reaction temperature,reaction p H,metal ion additives and substrate concentration was optimized,and the maximum c AMP production of 18.45 g·L^-1was achieved with a yield of 95.4%after bioconversion of 6 h.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2014AA022001)
文摘The unicellular green alga,Chlamydomonas reinhardtii is a model organism for studying various biological processes,such as photosynthesis,flagellar motility,and lipid metabolism.To find some novel genes regulating the lipid metabolism under various stress conditions,the paromomycin resistance gene aphVIII was transferred into the genome of C.reinhardtii to establish a mutant library.Two genes mutated in two of the TAG-reduced mutants(Cre06.g278111 in M2 mutant,Cre06.g278110 in M6 mutants)were neighboring in the genome,and their expression levels were down-regulated in their corresponding mutants in parallel with their reduced TAG levels following N deprivation.The proteins encoded by these two genes(KCN11 by Cre06.g278111,ACYC3 by Cre06.g278110)contained a conversed cyclic mononucleotide phosphate(cNMP)binding protein and an adenylate domain,respectively.Since cNMP binding protein and adenylate domain have been known as important components of cyclic adenosine monophosphate(cAMP)signaling pathway,suggesting that these two genes might af fect cellular TAG biosynthesis through cAMP signal pathway.
文摘To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role of hemin on this pathway, HTMCs of the third to fourth generation were cultured in vitro. Reverse transcripase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.
