Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purp...Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purpose of this study was to investigate the functions of CIRP in the testes. We employed RNAi technique to knock down the expression of CIRP in the testes, and performed haematoxylin and eosin staining to evaluate morphological changes following knockdown. Germ cell apoptosis was examined by terminal deoxynucleotidal transferase-mediated dUTP nick end labelling （TUNEL） assay, and mitogen-activated protein kinase （MAPK） signalling pathways were investigated by Western blotting to determine the possible mechanism of apoptosis. We found that using siRNA is a feasible and reliable method for knocking down gene expression in the testes. Compared to controls, the mean seminiferous tubule diameter （MSTD） and the thickness of the germ cell layers decreased following siRNA treatment, whereas the percentage of apoptotic seminiferous tubules increased. The p44/p42, p38 and SAPK/JNK MAPK pathways were activated after downregulation of CIRP. In conclusion, we discovered that downregulation of CIRP resulted in increased germ cell apoptosis, possibly viathe activation of the p44/p42, p38 and SAPK/JNK MAPK pathways.展开更多
To reveal the relationship between mitogen-acti-vated protein kinase (MAPK)and tau phosphorylation, we used different concentration of PD98059, an inhibitor of MEK (MAPKkinase), to treat mice neuroblastma (N2a) cell l...To reveal the relationship between mitogen-acti-vated protein kinase (MAPK)and tau phosphorylation, we used different concentration of PD98059, an inhibitor of MEK (MAPKkinase), to treat mice neuroblastma (N2a) cell line for 6 h. It showed that theactivity of MAPKdecreased in a dose-dependent manner. But Western blot and immunofluo-rescence revealed that justwhen the cells were treated with 16 mu mol/L PD98059, tau was hyperphosphorylated at Ser396/ 404 andSerl99/202 sites. We obtained the conclusion that overinhibited MAPK induced tauhyperphosphorylation at Ser396/404 and Serl99/202 sites.展开更多
Objective: To explore the effects of γ-irradiation on mitogen-activated protein kinases (MAPKs) and role of intracellular calcium in this event in intestinal epithelial cell line 6 (IEC-6 cells). Methods: After cultu...Objective: To explore the effects of γ-irradiation on mitogen-activated protein kinases (MAPKs) and role of intracellular calcium in this event in intestinal epithelial cell line 6 (IEC-6 cells). Methods: After cultured rat IIEC-6 cells with or without the pretreatment of intracellular Ca2+ chelator were exposed to Y-ir-radiation of 6 Gy, the total and phosphorylated MAPKs in the cells were determined with Western blotting and apoptosis was examined with flow cytometry. Activities of Extracellular signal-regulated protein kinase (ERK) and p38 MAPK were determined by using immuoprecipitation followed by Western blotting. Results: In response to γ-irradiation, phosphorylation of ERK was not significantly observed, while the levels of phosphorylated c-Jun NH2-terminal kinase (JNK) and p38 MAPK were increased in 30 min and reached the peak 2 h after exposure to 6 Gy γ-irradiation, though the cell viability was significantly lowered 12 h. On the other hand, no obvious changes were seen in the total protein levels of ERK, JNK and p38 MAPK. Chelation of intracellular Ca2+ almost completely suppressed the JNK and p38 MAPK phosphorylation induced by γ-irradia-tion, but removal of external Ca2+ had no such effect. Activation of p38 MAPK, but not of ERK, was seen to have a correlation with γ-irradiation induced apoptosis. Conclusion: The results suggest that γ-irradiation is a potent activator for JNK and p38 MAPK, and Ca2+ mobilized from intracellular stores plays an important role in the activation of MAPKs and the induction of apoptosis in IEC-6 cells.展开更多
An old drug with a new use can significantly reduce the cost and time for new drug research and development. MAPK （Mitogen-activated protein kinase） plays a very important key role in signal transduction pathways of...An old drug with a new use can significantly reduce the cost and time for new drug research and development. MAPK （Mitogen-activated protein kinase） plays a very important key role in signal transduction pathways of cell proliferation and differentiation. According to the statistics, there are about 30% persons who suffered from cancers related to the MAPK signal transduction pathways. Therefore, many researchers are focused on blocking these pathways in cancers therapies. Ras/Raf/MEK/ERK, however, is one of very important pathways among MAPK message transduction pathways. More and more information about MEK protein inhibitors are unveiled in several recent years. In the present study, the authors utilized MEK inhibitors which were already published and their activities were available to construct 2D-QSAR model by using CADD （multiple linear regression）. Then, the authors searched certified FDA drugs （Drugs@FDA 6184 drugs） making preliminary screening. The secondary screening on 3D structures were followed by using Docking, Scoring and Pharmacophore analysis to find out most suitable MEK inhibitors to become a fundamental database in drug discovery. The results are shown the ALogP, number of aromatic rings, number of hydrogen bond acceptors and number of hydrogen bond donors are all in positive correlation. According to the equation from 2D-QSAR model, the results conform to the previous description.展开更多
Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in ...Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in teleost.Identification,characterization,classification,and expression profiling of totally 15 mapk genes in black rockfish(Sebastes schlegelii)were conducted.Phylogenetic relationships show that these mapk genes could be divided into extracellular signal-regulated kinase(ERK),c-Jun N-terminal kinase(JNK),and p38 sub-families.In addition,gene structures,syntenic analysis,and selective pressure analysis are performed to confirm their annotations.Results of selective pressure analysis indicate that mapk1,mapk3,mapk7,mapk10,mapk11,and mapk12 underwent significantly-positive selections,while the others genes such as mapk4,mapk6,mapk15,mapk8a,mapk8b,mapk9,mapk13,mapk14a,and mapk14b were under purifying selections.Moreover,results of qRT-PCR indicate that mapk genes in 8 healthy tissues displayed different expression patterns.The expression patterns of several mapk genes including mapk12,mapk13,mapk14a,mapk14b,and mapk15 were significantly changed in mucosal tissues after Edwardsiella piscicida infection.This study demonstrates that mapk genes in black rockfish play vital prevention roles against bacterial infection,which not only helps us understand the structure and function of mapk genes in black rockfish,but also provides a reference to understand the role of mapk genes in teleost immune responses.展开更多
为研究异丙肾上腺素(isoproterenol,ISO)诱导心肌肥厚或心肌重塑的分子机制,本工作以成年雄性Balb/c小鼠为研究对象,通过腹腔注射ISO,采用蛋白免疫印迹杂交方法观察ISO对小鼠心肌丝裂素活化蛋白激酶(mitogen-activted protein kinase,MA...为研究异丙肾上腺素(isoproterenol,ISO)诱导心肌肥厚或心肌重塑的分子机制,本工作以成年雄性Balb/c小鼠为研究对象,通过腹腔注射ISO,采用蛋白免疫印迹杂交方法观察ISO对小鼠心肌丝裂素活化蛋白激酶(mitogen-activted protein kinase,MAPK)、核因子-κB(NFKB)和Janus激酶/信号转导因子和转录激活因子(JAK/STAT)途径的激活效应。结果发现,ISO腹腔注射后可早期(5 min)激活心肌MAPK(ERK1/2和p38);ISO对心肌NFκB的激活效应表现为双相性,激活高峰分别为5和120 min;ISO腹腔注射60 min后可显著促进STAT3的酪氨酸磷酸化,6h时基本恢复到基础水平。上述结果提示,ISO对多种细胞内信号转导途径均具有激活效应,但表现出明显的时相差异。探明这些信号转导途径的时空整合规律,将有助于深化对心肌重塑发生机制的认识。展开更多
MAPK(促分裂原活化蛋白激酶,Mitogen-Activated Protein Kinase)信号转导途径在植物病原真菌的生长发育、有性生殖和致病性调节等方面占有重要作用,是一种普遍存在的细胞外信号转导途径。在真核生物MAPK蛋白的同源性和2种炭疽菌(禾谷炭...MAPK(促分裂原活化蛋白激酶,Mitogen-Activated Protein Kinase)信号转导途径在植物病原真菌的生长发育、有性生殖和致病性调节等方面占有重要作用,是一种普遍存在的细胞外信号转导途径。在真核生物MAPK蛋白的同源性和2种炭疽菌(禾谷炭疽、希金斯炭疽菌)全基因组数据库的基础上,结合Blastp、蛋白质结构域分析和聚类分析3种方法,从禾谷炭疽菌和希金斯炭疽菌基因组数据库中找出18个与酿酒酵母菌同源的3类MAPK级联信号途径基因,并绘制出这2种炭疽菌MAPK级联信号途径简图,为深入研究炭疽菌MAPK级联信号途径基因生物学功能及其相关信号网络特点奠定基础。展开更多
文摘Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purpose of this study was to investigate the functions of CIRP in the testes. We employed RNAi technique to knock down the expression of CIRP in the testes, and performed haematoxylin and eosin staining to evaluate morphological changes following knockdown. Germ cell apoptosis was examined by terminal deoxynucleotidal transferase-mediated dUTP nick end labelling （TUNEL） assay, and mitogen-activated protein kinase （MAPK） signalling pathways were investigated by Western blotting to determine the possible mechanism of apoptosis. We found that using siRNA is a feasible and reliable method for knocking down gene expression in the testes. Compared to controls, the mean seminiferous tubule diameter （MSTD） and the thickness of the germ cell layers decreased following siRNA treatment, whereas the percentage of apoptotic seminiferous tubules increased. The p44/p42, p38 and SAPK/JNK MAPK pathways were activated after downregulation of CIRP. In conclusion, we discovered that downregulation of CIRP resulted in increased germ cell apoptosis, possibly viathe activation of the p44/p42, p38 and SAPK/JNK MAPK pathways.
文摘To reveal the relationship between mitogen-acti-vated protein kinase (MAPK)and tau phosphorylation, we used different concentration of PD98059, an inhibitor of MEK (MAPKkinase), to treat mice neuroblastma (N2a) cell line for 6 h. It showed that theactivity of MAPKdecreased in a dose-dependent manner. But Western blot and immunofluo-rescence revealed that justwhen the cells were treated with 16 mu mol/L PD98059, tau was hyperphosphorylated at Ser396/ 404 andSerl99/202 sites. We obtained the conclusion that overinhibited MAPK induced tauhyperphosphorylation at Ser396/404 and Serl99/202 sites.
基金in part by Natural Sciences Foundation of China (No. 39870239)by the Sasagawa Fellowship,Japan.
文摘Objective: To explore the effects of γ-irradiation on mitogen-activated protein kinases (MAPKs) and role of intracellular calcium in this event in intestinal epithelial cell line 6 (IEC-6 cells). Methods: After cultured rat IIEC-6 cells with or without the pretreatment of intracellular Ca2+ chelator were exposed to Y-ir-radiation of 6 Gy, the total and phosphorylated MAPKs in the cells were determined with Western blotting and apoptosis was examined with flow cytometry. Activities of Extracellular signal-regulated protein kinase (ERK) and p38 MAPK were determined by using immuoprecipitation followed by Western blotting. Results: In response to γ-irradiation, phosphorylation of ERK was not significantly observed, while the levels of phosphorylated c-Jun NH2-terminal kinase (JNK) and p38 MAPK were increased in 30 min and reached the peak 2 h after exposure to 6 Gy γ-irradiation, though the cell viability was significantly lowered 12 h. On the other hand, no obvious changes were seen in the total protein levels of ERK, JNK and p38 MAPK. Chelation of intracellular Ca2+ almost completely suppressed the JNK and p38 MAPK phosphorylation induced by γ-irradia-tion, but removal of external Ca2+ had no such effect. Activation of p38 MAPK, but not of ERK, was seen to have a correlation with γ-irradiation induced apoptosis. Conclusion: The results suggest that γ-irradiation is a potent activator for JNK and p38 MAPK, and Ca2+ mobilized from intracellular stores plays an important role in the activation of MAPKs and the induction of apoptosis in IEC-6 cells.
文摘An old drug with a new use can significantly reduce the cost and time for new drug research and development. MAPK （Mitogen-activated protein kinase） plays a very important key role in signal transduction pathways of cell proliferation and differentiation. According to the statistics, there are about 30% persons who suffered from cancers related to the MAPK signal transduction pathways. Therefore, many researchers are focused on blocking these pathways in cancers therapies. Ras/Raf/MEK/ERK, however, is one of very important pathways among MAPK message transduction pathways. More and more information about MEK protein inhibitors are unveiled in several recent years. In the present study, the authors utilized MEK inhibitors which were already published and their activities were available to construct 2D-QSAR model by using CADD （multiple linear regression）. Then, the authors searched certified FDA drugs （Drugs@FDA 6184 drugs） making preliminary screening. The secondary screening on 3D structures were followed by using Docking, Scoring and Pharmacophore analysis to find out most suitable MEK inhibitors to become a fundamental database in drug discovery. The results are shown the ALogP, number of aromatic rings, number of hydrogen bond acceptors and number of hydrogen bond donors are all in positive correlation. According to the equation from 2D-QSAR model, the results conform to the previous description.
基金Supported by the National Key R&D Program of China(No.2018YFD0900101)the Young Experts of Taishan Scholars(No.tsqn201909130)+2 种基金the Science and Technology Support Plan for Youth Innovation of Colleges and Universities in Shandong Province(No.2019KJF003)the“First Class Fishery Discipline”Program in Shandong Provincethe Shandong Technical System of Fish Industry(No.SDAIT-12-03)。
文摘Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in teleost.Identification,characterization,classification,and expression profiling of totally 15 mapk genes in black rockfish(Sebastes schlegelii)were conducted.Phylogenetic relationships show that these mapk genes could be divided into extracellular signal-regulated kinase(ERK),c-Jun N-terminal kinase(JNK),and p38 sub-families.In addition,gene structures,syntenic analysis,and selective pressure analysis are performed to confirm their annotations.Results of selective pressure analysis indicate that mapk1,mapk3,mapk7,mapk10,mapk11,and mapk12 underwent significantly-positive selections,while the others genes such as mapk4,mapk6,mapk15,mapk8a,mapk8b,mapk9,mapk13,mapk14a,and mapk14b were under purifying selections.Moreover,results of qRT-PCR indicate that mapk genes in 8 healthy tissues displayed different expression patterns.The expression patterns of several mapk genes including mapk12,mapk13,mapk14a,mapk14b,and mapk15 were significantly changed in mucosal tissues after Edwardsiella piscicida infection.This study demonstrates that mapk genes in black rockfish play vital prevention roles against bacterial infection,which not only helps us understand the structure and function of mapk genes in black rockfish,but also provides a reference to understand the role of mapk genes in teleost immune responses.
基金This work was supported by the Major State Research Development Program of People's Republic of China (No. G2000056906)the National Science Foundation of China (No. 30070872).
文摘为研究异丙肾上腺素(isoproterenol,ISO)诱导心肌肥厚或心肌重塑的分子机制,本工作以成年雄性Balb/c小鼠为研究对象,通过腹腔注射ISO,采用蛋白免疫印迹杂交方法观察ISO对小鼠心肌丝裂素活化蛋白激酶(mitogen-activted protein kinase,MAPK)、核因子-κB(NFKB)和Janus激酶/信号转导因子和转录激活因子(JAK/STAT)途径的激活效应。结果发现,ISO腹腔注射后可早期(5 min)激活心肌MAPK(ERK1/2和p38);ISO对心肌NFκB的激活效应表现为双相性,激活高峰分别为5和120 min;ISO腹腔注射60 min后可显著促进STAT3的酪氨酸磷酸化,6h时基本恢复到基础水平。上述结果提示,ISO对多种细胞内信号转导途径均具有激活效应,但表现出明显的时相差异。探明这些信号转导途径的时空整合规律,将有助于深化对心肌重塑发生机制的认识。
文摘MAPK(促分裂原活化蛋白激酶,Mitogen-Activated Protein Kinase)信号转导途径在植物病原真菌的生长发育、有性生殖和致病性调节等方面占有重要作用,是一种普遍存在的细胞外信号转导途径。在真核生物MAPK蛋白的同源性和2种炭疽菌(禾谷炭疽、希金斯炭疽菌)全基因组数据库的基础上,结合Blastp、蛋白质结构域分析和聚类分析3种方法,从禾谷炭疽菌和希金斯炭疽菌基因组数据库中找出18个与酿酒酵母菌同源的3类MAPK级联信号途径基因,并绘制出这2种炭疽菌MAPK级联信号途径简图,为深入研究炭疽菌MAPK级联信号途径基因生物学功能及其相关信号网络特点奠定基础。
