AIM:To examine the pathway related to the IL-1β-induced activation of mitogen-activated protein(MAP)kinases in cat esophageal smooth muscle cells.METHODS:Culture of the esophageal smooth musclecells from cat was prep...AIM:To examine the pathway related to the IL-1β-induced activation of mitogen-activated protein(MAP)kinases in cat esophageal smooth muscle cells.METHODS:Culture of the esophageal smooth musclecells from cat was prepared.Specific inhibitors weretreated before applying the IL-1β.Western blot analysiswas performed to detect the expressions of COX,iNOSand MAP kinases.RESULTS:In the primary cultured cells,although IL-1βfailed to upregulate the COX and iNOS levels,the levelsof the phosphorylated forms of p44/42 MAP kinase andp38 MAP kinase increased in both concentration-andtime-dependent manner,of which the level of activationreached a maximum within 3 and 18 h,respectively.The pertussis toxin reduced the level of p44/42 MAPkinase phosphorylation.Tyrphostin 51 and genistein alsoinhibited this activation.Neomycin decreased the densityof the p44/42 MAP kinase band to the basal level.Phosphokinase C(PKC)was found to play a mediatingrole in the IL-1β-induced p44/42 MAP kinase activity.In contrast,the activation of p38 MAP kinase wasinhibited only by a pretreatment with forskolin,and wasunaffected by the other compounds.CONCLUSION:Based on these results,IL-1β-inducedp44/42 MAP kinase activation is mediated by the Giprotein,tyrosine kinase,phospholipase C(PLC)andPKC.The pathway for p38 MAP kinase phosphorylationis different from that of p44/42 MAP kinase,suggestingthat it piays a different role in the cellular response to IL-1β.展开更多
Lung cancer has the highest mortality rate among all cancers,in part because it readily metastasizes.The tumor microenvironment,comprising blood vessels,fibroblasts,immune cells,and macrophages[including tumor-associa...Lung cancer has the highest mortality rate among all cancers,in part because it readily metastasizes.The tumor microenvironment,comprising blood vessels,fibroblasts,immune cells,and macrophages[including tumor-associated macrophages(TAMs)],is closely related to cancer cell growth,migration,and invasion.TAMs secrete several cytokines,including interleukin(IL)-1β,which participate in cancer migration and invasion.p21-activated kinase 1(PAK1),an important signaling molecule,induces cell migration and invasion in several carcinomas.Tonicityresponsive enhancer-binding protein(TonEBP)is also known to participate in cancer cell growth,migration,and invasion.However,the mechanisms by which it increases lung cancer migration remain unclear.Therefore,in this study,we aimed to elucidate the mechanisms by which IL-1βand TonEBP affect lung cancer cell migration and invasion.We found that A549 cocultured-MΦ-secreted IL-1βinduced A549 cell migration and invasion via the PAK1 pathway.TonEBP deficiency reduced A549 cell migration and invasion and increased responsiveness to IL-1β–induced migration and invasion.PAK1 phosphorylation,which was promoted by IL-1β,was reduced when TonEBP was depleted.These results suggest that TonEBP plays an important role in IL-1βinduction and invasiveness of A549 cells via the PAK1 pathway.These findings could be valuable in identifying potential targets for lung cancer treatment.展开更多
Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control...Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing serum),and then 24h after intervention respectively,the cell proliferation and Apoptosis rate;Western blot detected the expression levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65,MMP-13 proteins in chondrocytes.ELISA detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1βgroup;compared with IL-1βgroup,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell proliferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-αlevel decreased.13 protein levels and TNF-αand IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclusion:BSBT-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,improve the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.展开更多
文摘AIM:To examine the pathway related to the IL-1β-induced activation of mitogen-activated protein(MAP)kinases in cat esophageal smooth muscle cells.METHODS:Culture of the esophageal smooth musclecells from cat was prepared.Specific inhibitors weretreated before applying the IL-1β.Western blot analysiswas performed to detect the expressions of COX,iNOSand MAP kinases.RESULTS:In the primary cultured cells,although IL-1βfailed to upregulate the COX and iNOS levels,the levelsof the phosphorylated forms of p44/42 MAP kinase andp38 MAP kinase increased in both concentration-andtime-dependent manner,of which the level of activationreached a maximum within 3 and 18 h,respectively.The pertussis toxin reduced the level of p44/42 MAPkinase phosphorylation.Tyrphostin 51 and genistein alsoinhibited this activation.Neomycin decreased the densityof the p44/42 MAP kinase band to the basal level.Phosphokinase C(PKC)was found to play a mediatingrole in the IL-1β-induced p44/42 MAP kinase activity.In contrast,the activation of p38 MAP kinase wasinhibited only by a pretreatment with forskolin,and wasunaffected by the other compounds.CONCLUSION:Based on these results,IL-1β-inducedp44/42 MAP kinase activation is mediated by the Giprotein,tyrosine kinase,phospholipase C(PLC)andPKC.The pathway for p38 MAP kinase phosphorylationis different from that of p44/42 MAP kinase,suggestingthat it piays a different role in the cellular response to IL-1β.
基金the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-2014R1A6A1029617).
文摘Lung cancer has the highest mortality rate among all cancers,in part because it readily metastasizes.The tumor microenvironment,comprising blood vessels,fibroblasts,immune cells,and macrophages[including tumor-associated macrophages(TAMs)],is closely related to cancer cell growth,migration,and invasion.TAMs secrete several cytokines,including interleukin(IL)-1β,which participate in cancer migration and invasion.p21-activated kinase 1(PAK1),an important signaling molecule,induces cell migration and invasion in several carcinomas.Tonicityresponsive enhancer-binding protein(TonEBP)is also known to participate in cancer cell growth,migration,and invasion.However,the mechanisms by which it increases lung cancer migration remain unclear.Therefore,in this study,we aimed to elucidate the mechanisms by which IL-1βand TonEBP affect lung cancer cell migration and invasion.We found that A549 cocultured-MΦ-secreted IL-1βinduced A549 cell migration and invasion via the PAK1 pathway.TonEBP deficiency reduced A549 cell migration and invasion and increased responsiveness to IL-1β–induced migration and invasion.PAK1 phosphorylation,which was promoted by IL-1β,was reduced when TonEBP was depleted.These results suggest that TonEBP plays an important role in IL-1βinduction and invasiveness of A549 cells via the PAK1 pathway.These findings could be valuable in identifying potential targets for lung cancer treatment.
基金National Natural Science Foundation of China(No.82360934)Science and Technology Innovation Leading Talents Project of Xinjiang Uygur Autonomous Region(No.2022TSYCLJ0007)+1 种基金Xinjiang Uygur Autonomous Region Key Research and Development Task Special Project(No.2021B03006)Natural Science Foundat ion of Xinj iang Uygur Autonomous Region(No.2022D01C170,2022D01C171)。
文摘Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing serum),and then 24h after intervention respectively,the cell proliferation and Apoptosis rate;Western blot detected the expression levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65,MMP-13 proteins in chondrocytes.ELISA detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1βgroup;compared with IL-1βgroup,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell proliferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-αlevel decreased.13 protein levels and TNF-αand IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclusion:BSBT-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,improve the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.
