The nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage(SAH).WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions...The nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage(SAH).WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions,but its role in early brain injury(EBI)after SAH remains unclear.In this study,we used an in vivo SAH model in rats/mice and AAV-WNK1 intraventricular injection to investigate its neuroprotective mechanisms.WNK1 expression was significantly reduced in SAH patient blood and SAH model brain tissue,correlating negatively with microglial activation.AAV-WNK1 alleviated brain edema,neuronal necrosis,behavioral deficits,and inflammation by inhibiting NLRP3 inflammasome activation.In hemin-stimulated BV-2 cells,WNK1 overexpression reduced NLRP3 activation and inflammatory cytokines.Chloride counteracted WNK1’s inhibitory effects,and WNK1 suppressed P2X7R-induced NLRP3 activation.Mechanistically,WNK1 functioned via the OXSR1/STK39 pathway.These findings highlight WNK1 as a key regulator of intracellular chloride balance and neuroinflammation,presenting a potential therapeutic target for SAH treatment.展开更多
The association of polymorphisms in exon 1 of the WNK1 gene with essential hypertension in the minority groups of Hani and Yi of China was investigated in the case-control study.The sequence of 1257 bp containing the ...The association of polymorphisms in exon 1 of the WNK1 gene with essential hypertension in the minority groups of Hani and Yi of China was investigated in the case-control study.The sequence of 1257 bp containing the WNK1 gene exon 1 was determined in 1307 individuals（649 essential hypertension subjects and 658 controls）to identify SNPs in Hani and Yi minority groups.Four of eleven previously known SNPs （rs3168640,rs11885,rs11554421 and rs34880640）were identified.The SNP analysis indicated that SNPs rs11885 and rs11554421 were significantly associated with hypertension in both Hani and Yi populations,and rs34880640 was significantly associated with hypertension in Hani but not in Yi population,adjusted for covariates.Haplotype analysis indicated that the haplotype H1 significantly decreased the risk of hypertension in both populations.These results suggested that WNK1 polymorphisms were involved in the predisposition of essential hypertension in Hani and Yi populations and its effects showed a clear population specificity.This finding supported the importance of population specificity in determining the genetic factors associated with diseases and thus disease treatment.展开更多
Background:Damage to the corneal epithelium triggers important changes in the composition of the extracellular matrix(ECM)to which the basal human corneal epithelial cells(hCECs)attach.These changes are perceived by i...Background:Damage to the corneal epithelium triggers important changes in the composition of the extracellular matrix(ECM)to which the basal human corneal epithelial cells(hCECs)attach.These changes are perceived by integrins,a family of trans-membrane receptors that activate different intracellular signalling pathways,ultimately leading to re-epithelialization of the injured epithelium.Our goal is to study the impact of the pharmacological inhibition/activation of specific signal transduction mediators on corneal wound healing using both monolayers of hCECs and tissue-engineered human corneas(hTECs)as in vitro models.Methods:hTECs were produced by the self-assembly approach and wounded with a 8-mm biopsy punch.Total RNA and proteins were isolated from the wounded and unwounded hTECs to conduct gene profiling analyses and protein kinase arrays.The wounded tissues were then incubated either with the lysine deficient protein kinase 1(WNK1)inhibitor WNK463,the WNK1 indirect agonist AM1241,or with the vehicle alone(DMSO;negative control)and wound healing was monitored for 6 days.The impact of WNK1 inhibition/activation on hCECs monolayers was determined using scratch wound assays.Results:Gene profiling analyses and protein kinases arrays revealed that expression and activity of several mediators from the integrin-dependent signalling pathways were altered in response to the ECM changes taking place during corneal wound healing.Phosphorylation of the WNK1 kinase turned out to be the most striking activation event occurring during wound healing.Since the pharmacological inhibition of WNK1 by WNK463 significantly reduced the rate of corneal wound closure in our hTECs and hCECs monolayers compared to their respective negative controls,we believe that the pharmacological activation of WNK1 could turn out to be an interesting avenue to accelerate corneal wound closure.Conclusions:These results will contribute to a better understanding of the cellular and molecular mechanisms involved in corneal wound healing.Furthermore,they identified a new function for the WNK1 kinase in corneal wound healing and might lead to the identification of a new therapeutic target in the field of corneal wounds.展开更多
基金supported by Grants from the Basic Research Program of Jiangsu(BK20240492)Lianyungang Science and Technology Plan Project(JCYJ2304)+2 种基金Lianyungang Aging Health Research Project(L202301)Doctoral Research Start-up Fund of the First Peopel’Hospital of Lianyungang City(BS202314)The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘The nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome plays a crucial role in the prognosis of subarachnoid hemorrhage(SAH).WNK1 kinase negatively regulates NLRP3 in various inflammatory conditions,but its role in early brain injury(EBI)after SAH remains unclear.In this study,we used an in vivo SAH model in rats/mice and AAV-WNK1 intraventricular injection to investigate its neuroprotective mechanisms.WNK1 expression was significantly reduced in SAH patient blood and SAH model brain tissue,correlating negatively with microglial activation.AAV-WNK1 alleviated brain edema,neuronal necrosis,behavioral deficits,and inflammation by inhibiting NLRP3 inflammasome activation.In hemin-stimulated BV-2 cells,WNK1 overexpression reduced NLRP3 activation and inflammatory cytokines.Chloride counteracted WNK1’s inhibitory effects,and WNK1 suppressed P2X7R-induced NLRP3 activation.Mechanistically,WNK1 functioned via the OXSR1/STK39 pathway.These findings highlight WNK1 as a key regulator of intracellular chloride balance and neuroinflammation,presenting a potential therapeutic target for SAH treatment.
基金financially supported by the National Nature Science Foundation of China (Nos. U0932603 and 3091112048)the Natural Science Foundation of Yunnan Province (No. 2009CC001).
文摘The association of polymorphisms in exon 1 of the WNK1 gene with essential hypertension in the minority groups of Hani and Yi of China was investigated in the case-control study.The sequence of 1257 bp containing the WNK1 gene exon 1 was determined in 1307 individuals（649 essential hypertension subjects and 658 controls）to identify SNPs in Hani and Yi minority groups.Four of eleven previously known SNPs （rs3168640,rs11885,rs11554421 and rs34880640）were identified.The SNP analysis indicated that SNPs rs11885 and rs11554421 were significantly associated with hypertension in both Hani and Yi populations,and rs34880640 was significantly associated with hypertension in Hani but not in Yi population,adjusted for covariates.Haplotype analysis indicated that the haplotype H1 significantly decreased the risk of hypertension in both populations.These results suggested that WNK1 polymorphisms were involved in the predisposition of essential hypertension in Hani and Yi populations and its effects showed a clear population specificity.This finding supported the importance of population specificity in determining the genetic factors associated with diseases and thus disease treatment.
文摘Background:Damage to the corneal epithelium triggers important changes in the composition of the extracellular matrix(ECM)to which the basal human corneal epithelial cells(hCECs)attach.These changes are perceived by integrins,a family of trans-membrane receptors that activate different intracellular signalling pathways,ultimately leading to re-epithelialization of the injured epithelium.Our goal is to study the impact of the pharmacological inhibition/activation of specific signal transduction mediators on corneal wound healing using both monolayers of hCECs and tissue-engineered human corneas(hTECs)as in vitro models.Methods:hTECs were produced by the self-assembly approach and wounded with a 8-mm biopsy punch.Total RNA and proteins were isolated from the wounded and unwounded hTECs to conduct gene profiling analyses and protein kinase arrays.The wounded tissues were then incubated either with the lysine deficient protein kinase 1(WNK1)inhibitor WNK463,the WNK1 indirect agonist AM1241,or with the vehicle alone(DMSO;negative control)and wound healing was monitored for 6 days.The impact of WNK1 inhibition/activation on hCECs monolayers was determined using scratch wound assays.Results:Gene profiling analyses and protein kinases arrays revealed that expression and activity of several mediators from the integrin-dependent signalling pathways were altered in response to the ECM changes taking place during corneal wound healing.Phosphorylation of the WNK1 kinase turned out to be the most striking activation event occurring during wound healing.Since the pharmacological inhibition of WNK1 by WNK463 significantly reduced the rate of corneal wound closure in our hTECs and hCECs monolayers compared to their respective negative controls,we believe that the pharmacological activation of WNK1 could turn out to be an interesting avenue to accelerate corneal wound closure.Conclusions:These results will contribute to a better understanding of the cellular and molecular mechanisms involved in corneal wound healing.Furthermore,they identified a new function for the WNK1 kinase in corneal wound healing and might lead to the identification of a new therapeutic target in the field of corneal wounds.
