Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus.Angiogenesis is a major pathophysiology in endometriosis.Our previous studies have demonstrated that the pr...Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus.Angiogenesis is a major pathophysiology in endometriosis.Our previous studies have demonstrated that the prodrug of epigallocatechin gallate(ProEGCG)exhibits superior anti-endometriotic and anti-angiogenic effects compared to epigallocatechin gallate(EGCG).However,their direct binding targets and underlying mechanisms for the differential effects remain unknown.In this study,we demonstrated that oral ProEGCG can be effective in preventing and treating endometriosis.Additionally,1D and 2D Proteome Integral Solubility Alteration assay-based chemical proteomics identified metadherin(MTDH)and PX domain containing serine/threonine kinase-like(PXK)as novel binding targets of EGCG and ProEGCG,respectively.Computational simulation and BioLayer interferometry were used to confirm their binding affinity.Our results showed that MTDH-EGCG inhibited protein kinase B(Akt)-mediated angiogenesis,while PXK-ProEGCG inhibited epidermal growth factor(EGF)-mediated angiogenesis via the EGF/hypoxia-inducible factor(HIF-1a)/vascular endothelial growth factor(VEGF)pathway.In vitro and in vivo knockdown assays and microvascular network imaging further confirmed the involvement of these signaling pathways.Moreover,our study demonstrated that ProEGCG has superior therapeutic effects than EGCG by targeting distinct signal transduction pathways and may act as a novel antiangiogenic therapy for endometriosis.展开更多
Soluble receptor for advanced glycation end products(sRAGE)acts as a decoy sequestering of RAGE ligands,thus preventing the activation of the ligand-RAGE axis linking human diseases.However,the molecular mechanisms un...Soluble receptor for advanced glycation end products(sRAGE)acts as a decoy sequestering of RAGE ligands,thus preventing the activation of the ligand-RAGE axis linking human diseases.However,the molecular mechanisms underlying sRAGE remain unclear.In this study,THP-1 monocytes were cultured in normal glucose(NG,5.5 mmol/L)and high glucose(HG,15 mmol/L)to investigate the effects of diabetesrelevant glucose concentrations on sRAGE and interleukin-1β(IL-1β)secretion.The modulatory effects of epigallocatechin gallate(EGCG)in response to HG challenge were also evaluated.HG enhanced intracellular reactive oxygen species(ROS)generation and RAGE expression.The secretion of sRAGE,including esRAGE and cRAGE,was reduced under HG conditions,together with the downregulation of a disintegrin and metallopeptidase 10(ADAM10)and nuclear factor erythroid 2-related factor 2(Nrf2)nuclear translocation.Mechanistically,the HG effects were counteracted by siRAGE and exacerbated by siNrf2.Chromatin immunoprecipitation results showed that Nrf2 binding to the ADAM10 promoter and HG interfered with this binding.Our data reinforce the notion that RAGE and Nrf2 might be sRAGE-regulating factors.Under HG conditions,the treatment of EGCG reduced ROS generation and RAGE activation.EGCG-stimulated cRAGE release was likely caused by the upregulation of the Nrf2-ADAM10 pathway.EGCG inhibited HG-mediated NLRP3 inflammasome activation at least partly by stimulating sRAGE,thereby reducing IL-1βrelease.展开更多
为探究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)在乐果(dimethoate,DIM)经皮肤慢性暴露致小鼠骨骼肌损伤过程中是否具有保护作用,将6周龄雄性ICR小鼠随机分为对照组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L生理盐水...为探究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)在乐果(dimethoate,DIM)经皮肤慢性暴露致小鼠骨骼肌损伤过程中是否具有保护作用,将6周龄雄性ICR小鼠随机分为对照组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L生理盐水)、EGCG组(腹腔注射0.2 m L EGCG、背部涂抹0.1 m L生理盐水)、DIM组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L DIM)、DIM+EGCG组(腹腔注射0.2 m L EGCG、背部涂抹0.1 m L DIM),记录小鼠体重变化,检测小鼠骨骼肌MDA含量及CAT、T-SOD、T-AOC的活性,利用全自动生化分析仪检测血清CK和LDH活性,利用透射电镜观察骨骼肌超微结构,利用Western-blot检测Bip、PERK/eIF2α通路相关蛋白及成肌分化相关蛋白表达水平。结果显示,与对照组相比,DIM组第2~3周小鼠体重降低显著(P<0.05);第4~8周小鼠体重降低极显著(P<0.01);MDA含量升高极显著(P<0.01),CAT、T-SOD、T-AOC活性降低极显著(P<0.01);血清CK、LDH活性升高极显著(P<0.01);超微结构损伤;内质网应激标志蛋白Bip、PERK/eIF2α通路磷酸化蛋白表达水平显著升高(P<0.05);成肌分化关键蛋白的表达水平降低极显著(P<0.01)。与DIM组相比,DIM+EGCG组小鼠体重无显著差异,总体呈缓慢上升趋势;MDA含量降低极显著(P<0.01),CAT、T-SOD、T-AOC活性升高显著(P<0.05)或升高极显著(P<0.01);血清CK、LDH活性降低显著(P<0.05)或降低极显著(P<0.01);骨骼肌超微结构损伤有所缓解;Bip及PERK/eIF2α通路磷酸化蛋白表达水平降低显著(P<0.05)或降低极显著(P<0.01);成肌分化关键蛋白的表达水平升高显著(P<0.05)或升高极显著(P<0.01)。结果表明,DIM暴露导致了小鼠骨骼肌损伤,引起内质网应激并激活了下游的PERK/eIF2α通路,使小鼠骨骼肌成肌分化受损;EGCG能缓解DIM所致的氧化损伤,提高抗氧化能力,降低血清CK、LDH的活性,显著抑制内质网应激及下游PERK/eIF2α通路磷酸化水平,促进成肌分化蛋白的表达,改善DIM所致的损伤。展开更多
基金supported by the GRF RGC&CRF,Hong Kong(Grant Nos.:475012 and C5045-20 EF)HMRF,Hong Kong(Grant No.:03141386)+3 种基金ITF,Hong Kong(Grant No.:ITS/209/12)UGC Direct Grant 2011,2012,2021.032HKOG Trust Fund 2011,2014,2019the National Natural Science Foundation of China(Grant Nos.:81974225 and 82201823)。
文摘Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus.Angiogenesis is a major pathophysiology in endometriosis.Our previous studies have demonstrated that the prodrug of epigallocatechin gallate(ProEGCG)exhibits superior anti-endometriotic and anti-angiogenic effects compared to epigallocatechin gallate(EGCG).However,their direct binding targets and underlying mechanisms for the differential effects remain unknown.In this study,we demonstrated that oral ProEGCG can be effective in preventing and treating endometriosis.Additionally,1D and 2D Proteome Integral Solubility Alteration assay-based chemical proteomics identified metadherin(MTDH)and PX domain containing serine/threonine kinase-like(PXK)as novel binding targets of EGCG and ProEGCG,respectively.Computational simulation and BioLayer interferometry were used to confirm their binding affinity.Our results showed that MTDH-EGCG inhibited protein kinase B(Akt)-mediated angiogenesis,while PXK-ProEGCG inhibited epidermal growth factor(EGF)-mediated angiogenesis via the EGF/hypoxia-inducible factor(HIF-1a)/vascular endothelial growth factor(VEGF)pathway.In vitro and in vivo knockdown assays and microvascular network imaging further confirmed the involvement of these signaling pathways.Moreover,our study demonstrated that ProEGCG has superior therapeutic effects than EGCG by targeting distinct signal transduction pathways and may act as a novel antiangiogenic therapy for endometriosis.
文摘Soluble receptor for advanced glycation end products(sRAGE)acts as a decoy sequestering of RAGE ligands,thus preventing the activation of the ligand-RAGE axis linking human diseases.However,the molecular mechanisms underlying sRAGE remain unclear.In this study,THP-1 monocytes were cultured in normal glucose(NG,5.5 mmol/L)and high glucose(HG,15 mmol/L)to investigate the effects of diabetesrelevant glucose concentrations on sRAGE and interleukin-1β(IL-1β)secretion.The modulatory effects of epigallocatechin gallate(EGCG)in response to HG challenge were also evaluated.HG enhanced intracellular reactive oxygen species(ROS)generation and RAGE expression.The secretion of sRAGE,including esRAGE and cRAGE,was reduced under HG conditions,together with the downregulation of a disintegrin and metallopeptidase 10(ADAM10)and nuclear factor erythroid 2-related factor 2(Nrf2)nuclear translocation.Mechanistically,the HG effects were counteracted by siRAGE and exacerbated by siNrf2.Chromatin immunoprecipitation results showed that Nrf2 binding to the ADAM10 promoter and HG interfered with this binding.Our data reinforce the notion that RAGE and Nrf2 might be sRAGE-regulating factors.Under HG conditions,the treatment of EGCG reduced ROS generation and RAGE activation.EGCG-stimulated cRAGE release was likely caused by the upregulation of the Nrf2-ADAM10 pathway.EGCG inhibited HG-mediated NLRP3 inflammasome activation at least partly by stimulating sRAGE,thereby reducing IL-1βrelease.
文摘为探究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)在乐果(dimethoate,DIM)经皮肤慢性暴露致小鼠骨骼肌损伤过程中是否具有保护作用,将6周龄雄性ICR小鼠随机分为对照组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L生理盐水)、EGCG组(腹腔注射0.2 m L EGCG、背部涂抹0.1 m L生理盐水)、DIM组(腹腔注射0.2 m L生理盐水、背部涂抹0.1 m L DIM)、DIM+EGCG组(腹腔注射0.2 m L EGCG、背部涂抹0.1 m L DIM),记录小鼠体重变化,检测小鼠骨骼肌MDA含量及CAT、T-SOD、T-AOC的活性,利用全自动生化分析仪检测血清CK和LDH活性,利用透射电镜观察骨骼肌超微结构,利用Western-blot检测Bip、PERK/eIF2α通路相关蛋白及成肌分化相关蛋白表达水平。结果显示,与对照组相比,DIM组第2~3周小鼠体重降低显著(P<0.05);第4~8周小鼠体重降低极显著(P<0.01);MDA含量升高极显著(P<0.01),CAT、T-SOD、T-AOC活性降低极显著(P<0.01);血清CK、LDH活性升高极显著(P<0.01);超微结构损伤;内质网应激标志蛋白Bip、PERK/eIF2α通路磷酸化蛋白表达水平显著升高(P<0.05);成肌分化关键蛋白的表达水平降低极显著(P<0.01)。与DIM组相比,DIM+EGCG组小鼠体重无显著差异,总体呈缓慢上升趋势;MDA含量降低极显著(P<0.01),CAT、T-SOD、T-AOC活性升高显著(P<0.05)或升高极显著(P<0.01);血清CK、LDH活性降低显著(P<0.05)或降低极显著(P<0.01);骨骼肌超微结构损伤有所缓解;Bip及PERK/eIF2α通路磷酸化蛋白表达水平降低显著(P<0.05)或降低极显著(P<0.01);成肌分化关键蛋白的表达水平升高显著(P<0.05)或升高极显著(P<0.01)。结果表明,DIM暴露导致了小鼠骨骼肌损伤,引起内质网应激并激活了下游的PERK/eIF2α通路,使小鼠骨骼肌成肌分化受损;EGCG能缓解DIM所致的氧化损伤,提高抗氧化能力,降低血清CK、LDH的活性,显著抑制内质网应激及下游PERK/eIF2α通路磷酸化水平,促进成肌分化蛋白的表达,改善DIM所致的损伤。
