Recurrent pregnancy loss （RPL） is a condition with complex etiologies, to which both genetic and envi- ronmental factors may contribute. During the last decade, studies indicated that the expression patterns of the ...Recurrent pregnancy loss （RPL） is a condition with complex etiologies, to which both genetic and envi- ronmental factors may contribute. During the last decade, studies indicated that the expression patterns of the pro- kineticin receptor （PKR1 and PKR2） are closely related to early pregnancy. However, there are few studies on the role of PKR1 and PKR2 in RPL. In this study, we purpose to investigate the association between polymorphisms of the prokineticin receptor （PKR1 rs4627609 and PKR2 rs6053283） and RPL on a group of 93 RPL cases and 169 healthy controls. Genotyping of the single nucleotide polymorphisms （SNPs） was performed using a Sequenom MassARRAY iPLEX system. The results revealed a significant association between PKR2 rs6053283 polymorphism and RPL （P=0.003）, whereas no association was observed between PKR1 rs4627609 polymorphism and RPL （P=-0.929） in the Chinese Han population.展开更多
The interferon-inducible double-stranded RNA-dependent protein kinase PKR has been suggested to function as a turnour suppressor gene product.Catalytically inactive mutants of PKR give rise to a tumourigenic phenotype...The interferon-inducible double-stranded RNA-dependent protein kinase PKR has been suggested to function as a turnour suppressor gene product.Catalytically inactive mutants of PKR give rise to a tumourigenic phenotype when overexpressed in NIH-3T3 fibroblasts and this has been attributed to a dominant negative effect on only inhibits the protein kinase activity of wild-type PKR but is also inhibitory towards another dotlblestranded RNA-dependent enzyme,the 40kDa form of 2'5'oligoadenylate synthetase. Inhibition of both wile-type PKR or 2'5' oligoadenylate synthetase is reversed by adding higher concentrations of double-stranded RNA. These results suggest competition between PKR K296R and wild-type PKR or 2'5' oligoadenylate synthetase for limiting amounts of dublestranded RNA. Moreover,the data imply that the tumourigenic effect of this PKR mutant could be due to inhibition of additional pathways requiring low levels of double-strandeed RNA for activation and cannot be unambiguously attributed to inhibition of endogenous PKR itself.展开更多
Objective:To observe the effect of Sanshi decoction on P2X7R/PKR pathway-mediated activation of macrophage NLRP3 inflammasome to elucidate the molecular mechanism of Sanshi decoction in the treatment of gouty arthriti...Objective:To observe the effect of Sanshi decoction on P2X7R/PKR pathway-mediated activation of macrophage NLRP3 inflammasome to elucidate the molecular mechanism of Sanshi decoction in the treatment of gouty arthritis.Methods:THP-1 macrophages were divided into control group,model group,low dose group,medium dose group,high dose group of Sanshi decoction and inhibitor group.The remaining groups were induced with monosodium urate crystals to establish a gouty arthritis cell model except the control group.Flow cytometry was used to detect macrophage ROS levels in each group,ELISA to detect MDA levels and SOD and GSH-PX activities in each group,and Western blot to detect P2X7R/PKR pathway and NLRP3 inflammasome-associated protein expression.We also used CCK-8 and flow cytometry to measure MH7A activity and apoptotic levels.Results:Compared with the control group,the ROS level,the content of MDA,the activities of SOD and GSH-PX were significantly increased,and the expression levels of NLRP3,full-length IL-1β,pro-IL-1β,full-length IL-18,pro-IL-18,full-length caspase-1,GSDMD-NT,P2X7R and p-PKR protein expression levels were significantly upregulated,and GSDMD-FL protein expression was significantly downregulated in the model group,and that the differences between them were statistically significant(P<0.05 and P<0.01).Compared with the model group,Sanshi decoction could reduce macrophage ROS levels,MDA content,SOD and GSHPX activities,and downregulate macrophage NLRP3,mature IL-1β,pro IL-1β,mature IL-18,pro IL-18,mature caspase-1,GSDMD-NT,P2X7R and p-PKR protein expression,and upregulate GSDMD-FL protein expression,with statistically significant differences(P<0.05 and P<0.01).In addition,MH7A activity was downregulated,and apoptosis level was upregulated in the model group in comparison with the control group,and differences were all significantly different(P<0.05).As compared to the model group,Sanshi decoction could significantly increase the activity of MH7A and inhibit the level of apoptosis,and that the differences between them were statistically significant(P<0.05 and P<0.01).Conclusion:Sanshi decoction can achieve the therapeutic effect of gouty arthritis by inhibiting P2X7R/PKR pathway activation,thus reducing the activation level of NLRP3.展开更多
基金Project supported by the National Natural Science Foundation of China(No.81571503)
文摘Recurrent pregnancy loss （RPL） is a condition with complex etiologies, to which both genetic and envi- ronmental factors may contribute. During the last decade, studies indicated that the expression patterns of the pro- kineticin receptor （PKR1 and PKR2） are closely related to early pregnancy. However, there are few studies on the role of PKR1 and PKR2 in RPL. In this study, we purpose to investigate the association between polymorphisms of the prokineticin receptor （PKR1 rs4627609 and PKR2 rs6053283） and RPL on a group of 93 RPL cases and 169 healthy controls. Genotyping of the single nucleotide polymorphisms （SNPs） was performed using a Sequenom MassARRAY iPLEX system. The results revealed a significant association between PKR2 rs6053283 polymorphism and RPL （P=0.003）, whereas no association was observed between PKR1 rs4627609 polymorphism and RPL （P=-0.929） in the Chinese Han population.
文摘The interferon-inducible double-stranded RNA-dependent protein kinase PKR has been suggested to function as a turnour suppressor gene product.Catalytically inactive mutants of PKR give rise to a tumourigenic phenotype when overexpressed in NIH-3T3 fibroblasts and this has been attributed to a dominant negative effect on only inhibits the protein kinase activity of wild-type PKR but is also inhibitory towards another dotlblestranded RNA-dependent enzyme,the 40kDa form of 2'5'oligoadenylate synthetase. Inhibition of both wile-type PKR or 2'5' oligoadenylate synthetase is reversed by adding higher concentrations of double-stranded RNA. These results suggest competition between PKR K296R and wild-type PKR or 2'5' oligoadenylate synthetase for limiting amounts of dublestranded RNA. Moreover,the data imply that the tumourigenic effect of this PKR mutant could be due to inhibition of additional pathways requiring low levels of double-strandeed RNA for activation and cannot be unambiguously attributed to inhibition of endogenous PKR itself.
基金Heilongjiang Province Traditional Chinese Medicine Research Project(No.ZHY19-006)。
文摘Objective:To observe the effect of Sanshi decoction on P2X7R/PKR pathway-mediated activation of macrophage NLRP3 inflammasome to elucidate the molecular mechanism of Sanshi decoction in the treatment of gouty arthritis.Methods:THP-1 macrophages were divided into control group,model group,low dose group,medium dose group,high dose group of Sanshi decoction and inhibitor group.The remaining groups were induced with monosodium urate crystals to establish a gouty arthritis cell model except the control group.Flow cytometry was used to detect macrophage ROS levels in each group,ELISA to detect MDA levels and SOD and GSH-PX activities in each group,and Western blot to detect P2X7R/PKR pathway and NLRP3 inflammasome-associated protein expression.We also used CCK-8 and flow cytometry to measure MH7A activity and apoptotic levels.Results:Compared with the control group,the ROS level,the content of MDA,the activities of SOD and GSH-PX were significantly increased,and the expression levels of NLRP3,full-length IL-1β,pro-IL-1β,full-length IL-18,pro-IL-18,full-length caspase-1,GSDMD-NT,P2X7R and p-PKR protein expression levels were significantly upregulated,and GSDMD-FL protein expression was significantly downregulated in the model group,and that the differences between them were statistically significant(P<0.05 and P<0.01).Compared with the model group,Sanshi decoction could reduce macrophage ROS levels,MDA content,SOD and GSHPX activities,and downregulate macrophage NLRP3,mature IL-1β,pro IL-1β,mature IL-18,pro IL-18,mature caspase-1,GSDMD-NT,P2X7R and p-PKR protein expression,and upregulate GSDMD-FL protein expression,with statistically significant differences(P<0.05 and P<0.01).In addition,MH7A activity was downregulated,and apoptosis level was upregulated in the model group in comparison with the control group,and differences were all significantly different(P<0.05).As compared to the model group,Sanshi decoction could significantly increase the activity of MH7A and inhibit the level of apoptosis,and that the differences between them were statistically significant(P<0.05 and P<0.01).Conclusion:Sanshi decoction can achieve the therapeutic effect of gouty arthritis by inhibiting P2X7R/PKR pathway activation,thus reducing the activation level of NLRP3.
