BACKGROUND Periodontitis is an inflammatory disease caused by the host’s immune response and various interactions between pathogens,which lead to the loss of connective tissue and bone.In recent years,mesenchymal ste...BACKGROUND Periodontitis is an inflammatory disease caused by the host’s immune response and various interactions between pathogens,which lead to the loss of connective tissue and bone.In recent years,mesenchymal stem cell(SC)transplantation technology has become a research hotspot,which can form periodontal ligament,cementum,and alveolar bone through proliferation and differentiation.AIM To elucidate the regulatory effects of WD repeat-containing protein 36(WDR36)on the senescence,migration,and osteogenic differentiation of periodontal ligament SCs(PDLSCs).METHODS The migration and chemotaxis of PDLSCs were detected by the scratch-wound migration test and transwell chemotaxis test.Alkaline phosphatase(ALP)activity,Alizarin red staining,calcium content,and real-time reverse transcription polymerase chain reaction(RT-qPCR)of key transcription factors were used to detect the osteogenic differentiation function of PDLSCs.Cell senescence was determined by senescence-associatedβ-galactosidase staining.RESULTS The 24-hour and 48-hour scratch-wound migration test and 48-hour transwell chemotaxis test showed that overexpression of WDR36 inhibited the migration/chemotaxis of PDLSCs.Simultaneously,WDR36 depletion promoted the migration/chemotaxis of PDLSCs.The results of ALP activity,Alizarin red staining,calcium content,and RTqPCR showed that overexpression of WDR36 inhibited the osteogenic differentiation of PDLSCs,and WDR36 depletion promoted the osteogenic differentiation of PDLSCs.Senescence-associatedβ-galactosidase staining showed that 0.1μg/mL icariin(ICA)and overexpression of WDR36 inhibited the senescence of PDLSCs,and WDR36 depletion promoted the osteogenic differentiation of PDLSCs.CONCLUSION WDR36 inhibits the migration and chemotaxis,osteogenic differentiation,and senescence of PDLSCs;0.1μg/mL ICA inhibits the senescence of PDLSCs.Therefore,WDR36 might serve as a target for periodontal tissue regeneration and the treatment of periodontitis.展开更多
BACKGROUND The mortality rate for severe cases of acute pancreatitis(AP),a common gastrointestinal emergency,is as high as 30%.Our previous study has shown that rutaecarpine(Rut)has a therapeutic effect on AP.AIM To i...BACKGROUND The mortality rate for severe cases of acute pancreatitis(AP),a common gastrointestinal emergency,is as high as 30%.Our previous study has shown that rutaecarpine(Rut)has a therapeutic effect on AP.AIM To investigate the role of F-box and WD repeat domain containing 11(FBXW11)in AP models and to assess whether Rut mitigates AP by regulating FBXW11.METHODS AP rat model was established and treated with Rut,followed by biochemical analysis of serum amylase and lipase,hematoxylin and eosin staining of pancreatic tissue,and immunohistochemistry detection of pancreatic Ly6G,CD11b,and myeloperoxidase.Assay kits were used to detect oxidative stress-related indicators in pancreatic tissue and inflammatory factors in serum.AR42J cells were treated with cerulein to model AP and subjected to Cell Counting Kit-8 viability assay,flow cytometry apoptosis assay,and immunofluorescence detection of reactive oxygen species to elucidate the mechanistic involvement of the enhancer of zeste homolog 2(EZH2)-FBXW11 axis in Rut-mediated protection against AP.The EZH2-histone H3 binding and H3 methylation were evaluated using co-immunoprecipitation.RESULTS Rut treatment ameliorated AP severity,as evidenced by reduced serum levels of pancreatic enzymes(amylase and lipase)and attenuated histological damage.Rut also decreased inflammatory markers(interleukin-1 beta,interleukin-6,and tumor necrosis factor alpha),tissue oxidative stress(malondialdehyde),and neutrophil infiltration(Ly6G,CD11b,and myeloperoxidase)levels in rats with AP.Moreover,Rut restored pancreatic antioxidant capacity(glutathione and superoxide dismutase).In vitro,Rut pre-incubation enhanced cell viability and suppressed cerulein-induced apoptosis and oxidative stress.Rut increased EZH2 expression while decreasing FBXW11 expression.FBXW11 overexpression eliminated the protective effect of Rut against AP.Further analysis revealed that EZH2 binds to H3 and upregulates H3 methylation levels,thereby inhibiting FBXW11 expression.CONCLUSION Collectively,our findings demonstrate that Rut ameliorates AP by upregulating EZH2,thereby enhancing H3 methylation and suppressing FBXW11 expression.展开更多
文摘BACKGROUND Periodontitis is an inflammatory disease caused by the host’s immune response and various interactions between pathogens,which lead to the loss of connective tissue and bone.In recent years,mesenchymal stem cell(SC)transplantation technology has become a research hotspot,which can form periodontal ligament,cementum,and alveolar bone through proliferation and differentiation.AIM To elucidate the regulatory effects of WD repeat-containing protein 36(WDR36)on the senescence,migration,and osteogenic differentiation of periodontal ligament SCs(PDLSCs).METHODS The migration and chemotaxis of PDLSCs were detected by the scratch-wound migration test and transwell chemotaxis test.Alkaline phosphatase(ALP)activity,Alizarin red staining,calcium content,and real-time reverse transcription polymerase chain reaction(RT-qPCR)of key transcription factors were used to detect the osteogenic differentiation function of PDLSCs.Cell senescence was determined by senescence-associatedβ-galactosidase staining.RESULTS The 24-hour and 48-hour scratch-wound migration test and 48-hour transwell chemotaxis test showed that overexpression of WDR36 inhibited the migration/chemotaxis of PDLSCs.Simultaneously,WDR36 depletion promoted the migration/chemotaxis of PDLSCs.The results of ALP activity,Alizarin red staining,calcium content,and RTqPCR showed that overexpression of WDR36 inhibited the osteogenic differentiation of PDLSCs,and WDR36 depletion promoted the osteogenic differentiation of PDLSCs.Senescence-associatedβ-galactosidase staining showed that 0.1μg/mL icariin(ICA)and overexpression of WDR36 inhibited the senescence of PDLSCs,and WDR36 depletion promoted the osteogenic differentiation of PDLSCs.CONCLUSION WDR36 inhibits the migration and chemotaxis,osteogenic differentiation,and senescence of PDLSCs;0.1μg/mL ICA inhibits the senescence of PDLSCs.Therefore,WDR36 might serve as a target for periodontal tissue regeneration and the treatment of periodontitis.
基金Supported by Hunan Provincial Natural Science Foundation of China,No.2022JJ40836the Key Project of Research and Development Plan of Hunan Province,No.2023DK2002the Postdoctoral Fellowship Program of China Postdoctoral Science Foundation,No.GZC20242045.
文摘BACKGROUND The mortality rate for severe cases of acute pancreatitis(AP),a common gastrointestinal emergency,is as high as 30%.Our previous study has shown that rutaecarpine(Rut)has a therapeutic effect on AP.AIM To investigate the role of F-box and WD repeat domain containing 11(FBXW11)in AP models and to assess whether Rut mitigates AP by regulating FBXW11.METHODS AP rat model was established and treated with Rut,followed by biochemical analysis of serum amylase and lipase,hematoxylin and eosin staining of pancreatic tissue,and immunohistochemistry detection of pancreatic Ly6G,CD11b,and myeloperoxidase.Assay kits were used to detect oxidative stress-related indicators in pancreatic tissue and inflammatory factors in serum.AR42J cells were treated with cerulein to model AP and subjected to Cell Counting Kit-8 viability assay,flow cytometry apoptosis assay,and immunofluorescence detection of reactive oxygen species to elucidate the mechanistic involvement of the enhancer of zeste homolog 2(EZH2)-FBXW11 axis in Rut-mediated protection against AP.The EZH2-histone H3 binding and H3 methylation were evaluated using co-immunoprecipitation.RESULTS Rut treatment ameliorated AP severity,as evidenced by reduced serum levels of pancreatic enzymes(amylase and lipase)and attenuated histological damage.Rut also decreased inflammatory markers(interleukin-1 beta,interleukin-6,and tumor necrosis factor alpha),tissue oxidative stress(malondialdehyde),and neutrophil infiltration(Ly6G,CD11b,and myeloperoxidase)levels in rats with AP.Moreover,Rut restored pancreatic antioxidant capacity(glutathione and superoxide dismutase).In vitro,Rut pre-incubation enhanced cell viability and suppressed cerulein-induced apoptosis and oxidative stress.Rut increased EZH2 expression while decreasing FBXW11 expression.FBXW11 overexpression eliminated the protective effect of Rut against AP.Further analysis revealed that EZH2 binds to H3 and upregulates H3 methylation levels,thereby inhibiting FBXW11 expression.CONCLUSION Collectively,our findings demonstrate that Rut ameliorates AP by upregulating EZH2,thereby enhancing H3 methylation and suppressing FBXW11 expression.
