AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein e...AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein endothelial cells(HUVECs)to induce hypoxia in vitro.Eight-week-old male C57 BL/6 J mice weighing 19-25 g were used for a choroidal neovascularization(CNV)model induced by laser photocoagulation in vivo.Expression levels of YAP,phosphorylated YAP,mesenchymal markers[αsmooth muscle actin(α-SMA),vimentin,and Snail],and endothelial cell markers(CD31 and zonula occludens 1)were measured by Western blotting,quantitative real-time PCR,and immunofluorescence microscopy.Small molecules YC-1(Lificiguat,a specific inhibitor of hypoxia-inducible factor 1α),CA3(CIL56,an inhibitor of YAP),and XMU-MP-1(an inhibitor of Hippo kinase MST1/2,which activates YAP)were used to explore the underlying mechanism.RESULTS:Co Cl2 increased expression of mesenchymal markers,decreased expression of endothelial cell markers,and enhanced the ability of primary HUVECs to proliferate and migrate.YC-1 suppressed hypoxia-induced endothelialto-mesenchymal transition(End MT).Moreover,hypoxia promoted total expression,inhibited phosphorylation,and enhanced the transcriptional activity of YAP.XMU-MP-1 enhanced hypoxia-induced End MT,whereas CA3 elicited the opposite effect.Expression of YAP,α-SMA,and vimentin were upregulated in the laser-induced CNV model.However,silencing of YAP by vitreous injection of small interfering RNA targeting YAP could reverse these changes.CONCLUSION:The findings reveal a critical role of the hypoxia-inducible factor-1α(HIF-1α)/YAP signaling axis in End MT and provide a new therapeutic target for treatment of subretinal fibrosis in AMD.展开更多
Pulmonary hypertension (PH) is a fatal disorder characterized by pulmonary vascular remodeling and obstruction. The phosphodiesterase 4 (PDE4) family hydrolyzes cyclic AMP (cAMP) and is comprised of four subtypes (PD...Pulmonary hypertension (PH) is a fatal disorder characterized by pulmonary vascular remodeling and obstruction. The phosphodiesterase 4 (PDE4) family hydrolyzes cyclic AMP (cAMP) and is comprised of four subtypes (PDE4A–D). Previous studies have shown the beneficial effects of pan-PDE4 inhibitors in rodent PH;however, this class of drugs is associated with side effects owing to the broad inhibition of all four PDE4 isozymes. Here, we demonstrate that PDE4B is the predominant PDE isozyme in lungs and that it was upregulated in rodent and human PH lung tissues. We also confirmed that PDE4B is mainly expressed in the lung endothelial cells (ECs). Evaluation of PH in Pde4b wild type and knockout mice confirmed that Pde4b is important for the vascular remodeling associated with PH. In vivo EC lineage tracing demonstrated that Pde4b induces PH development by driving endothelial-to-mesenchymal transition (EndMT), and mechanistic studies showed that Pde4b regulates EndMT by antagonizing the cAMP-dependent PKA–CREB–BMPRII axis. Finally, treating PH rats with a PDE4B-specific inhibitor validated that PDE4B inhibition has a significant pharmacological effect in the alleviation of PH. Collectively, our findings indicate a critical role for PDE4B in EndMT and PH, prompting further studies of PDE4B-specific inhibitors as a therapeutic strategy for PH.展开更多
Glycidol is a common lipid-derived foodborne toxicant mainly presents in refined oils and related foodstuffs.Vascular endothelial cells may be potential targets of the deleterious effects associated with glycidol expo...Glycidol is a common lipid-derived foodborne toxicant mainly presents in refined oils and related foodstuffs.Vascular endothelial cells may be potential targets of the deleterious effects associated with glycidol exposure.In human umbilical vein endothelial cells(HUVECs),we found that glycidol treatment promoted endothelialto-mesenchymal transition(EndMT)at a lower concentration(0.5 mmol/L),while induced apoptosis and inflammation at a higher concentration(1 mmol/L).These harmful effects were achieved by the activation of NF-κB/MAPK signaling pathway and were mediated by reactive oxygen species(ROS).In addition,the protective potential of 6-C-(E-2-fluorostyryl)naringenin(6-CEFN)against glycidol was evaluated and compared with naringenin.HUVECs pre-treated with 6-CEFN,but not naringenin,displayed resistance to endothelial dysfunction caused by glycidol.展开更多
Objective:To investigate whether Leech-Centipede(LC) Granules can improve erectile function in rats with diabetes mellitus-associated erectile dysfunction(DMED) through endothelial-to-mesenchymal transition(EndMT) inh...Objective:To investigate whether Leech-Centipede(LC) Granules can improve erectile function in rats with diabetes mellitus-associated erectile dysfunction(DMED) through endothelial-to-mesenchymal transition(EndMT) inhibition.Methods:Components of LC Granules were identified via ultra-high-performance liquid chromatography.Thirty male Sprague Dawley rats were injected with streptozotocin and fed continuously for 8weeks to establish the DMED rat model.Rats with erectile dysfunction symptoms diagnosed using apomorphine were divided into DMED and low-,medium-,and high-doses LC groups(n=6 in each).The negative control(NC, n=6) and DMED groups were given 5 mL of deionized water via intragastric gavage,and the low-,mediumand the high-doses LC groups were administered LC at 1.6,3.2,and 6.4 g/kg,respectively,via intragastric gavage for 4 weeks.The intracavernous pressure(ICP),mean arterial pressure(MAP),and nitric oxide(NO) levels in cavernous tissue were measured for each group.Quantitative reverse transcription-polymerase chain reaction and Western blot were used to detect mRNA and protein expressions of endothelial and mesenchymal markers.Immunofluorescence staining was used to observe α-SMA, and Masson’s trichrome staining was performed to determine the myofiber/collagen ratio.Results:A total of 474 active components were identified.After treatment,the ICP/MAP value and NO level were significantly higher in the medium-and high-dose LC groups than in the DMED group(P<0.05).Compared with the DMED groups,the medium-and high-dose groups LC significantly increased and decreased endothelial and mesenchymal markers expression,respectively(P<0.05).Tumor growth factor(TGF) β RⅡ,p-Smad2, and p-Smad3 levels were considerably higher following diabetes onset but reduced following LC intervention(P<0.05),except for TGF β 1(P>0.05).α-SMA expression was significantly higher in the DMED group and was reduced in all LC intervention groups(P>0.05).The myofiber/collagen ratio in the LC groups was higher than that in the DMED group but lower than that in the NC group(all P<0.05).Conclusions:LC Granules may improve the erectile function of DMED rats by suppressing TGF-β/Smad pathway to reverse EndMT.展开更多
Small-diameter tissue-engineered vascular grafts(sdTEVGs)have garnered significant attention as a potential treatment modality for vascular bypass grafting and replacement therapy.However,the intimal hyperplasia and t...Small-diameter tissue-engineered vascular grafts(sdTEVGs)have garnered significant attention as a potential treatment modality for vascular bypass grafting and replacement therapy.However,the intimal hyperplasia and thrombosis are two major complications that impair graft patency during transplantation.To address this issue,we fabricated the covalent-organic framework(COF)-based carbon monoxide(CO)nanogenerator-and co-immobilized with LXW-7 peptide and heparin to establish a multifunctional surface on TEVGs constructed from acellular blood vessels for preventing thrombosis and stenosis.The cell-adhesive peptide LXW-7 could capture endothelial-forming cells(EFCs)to promote endothelialization,while the antithrombotic molecule heparin prevented thrombus formation.The reactive oxygen species(ROS)-triggered CO release suppressed the adhesion and activation of macrophages,leading to the reduction of ROS and inflammatory factors.As a result,the endothelial-to-mesenchymal transition(EndMT)triggered by inflammation was restricted,facilitating the maintenance of the homeostasis of the neo-endothelium and preventing pathological remodeling in TEVGs.When transplanted in vivo,these vascular grafts exhibited negligible intimal hyperplasia and remained patent for 3 months.This achievement provided a novel approach for constructing antithrombotic and anti-hyperplastic TEVGs.展开更多
基金National Natural Science Foundation of China(No.81970817,No.81873680)。
文摘AIM:To reveal whether and how Yes-associated protein(YAP)promotes the occurrence of subretinal fibrosis in agerelated macular degeneration(AMD).METHODS:Cobalt chloride(Co Cl2)was used in primary human umbilical vein endothelial cells(HUVECs)to induce hypoxia in vitro.Eight-week-old male C57 BL/6 J mice weighing 19-25 g were used for a choroidal neovascularization(CNV)model induced by laser photocoagulation in vivo.Expression levels of YAP,phosphorylated YAP,mesenchymal markers[αsmooth muscle actin(α-SMA),vimentin,and Snail],and endothelial cell markers(CD31 and zonula occludens 1)were measured by Western blotting,quantitative real-time PCR,and immunofluorescence microscopy.Small molecules YC-1(Lificiguat,a specific inhibitor of hypoxia-inducible factor 1α),CA3(CIL56,an inhibitor of YAP),and XMU-MP-1(an inhibitor of Hippo kinase MST1/2,which activates YAP)were used to explore the underlying mechanism.RESULTS:Co Cl2 increased expression of mesenchymal markers,decreased expression of endothelial cell markers,and enhanced the ability of primary HUVECs to proliferate and migrate.YC-1 suppressed hypoxia-induced endothelialto-mesenchymal transition(End MT).Moreover,hypoxia promoted total expression,inhibited phosphorylation,and enhanced the transcriptional activity of YAP.XMU-MP-1 enhanced hypoxia-induced End MT,whereas CA3 elicited the opposite effect.Expression of YAP,α-SMA,and vimentin were upregulated in the laser-induced CNV model.However,silencing of YAP by vitreous injection of small interfering RNA targeting YAP could reverse these changes.CONCLUSION:The findings reveal a critical role of the hypoxia-inducible factor-1α(HIF-1α)/YAP signaling axis in End MT and provide a new therapeutic target for treatment of subretinal fibrosis in AMD.
基金This work was supported by Beijing Natural Science Foundation[Z220019 to Jing Wang,China]National High Level of Hospital Clinical Research Funding[2022-PUMCH-D-002 to Jing Wang,China]+3 种基金National Key Research and Development Program of China Grants[2019YFA0801703 and 2019YFA0801804 to Jing Wang]Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences[2022-I2M-JB-007 to Chen Wang,2021-I2M-1-016 to Hongmei Zhao,2021-I2M-1-049 to Jing Wang,2021-I2M-1-005 to Yanjiang Xing,China]Haihe Laboratory of Cell Ecosystem Innovation Fund[22HHXBSS00010 to Jing Wang,China]National Natural Science Foundation of China[82241004 to Jing Wang].
文摘Pulmonary hypertension (PH) is a fatal disorder characterized by pulmonary vascular remodeling and obstruction. The phosphodiesterase 4 (PDE4) family hydrolyzes cyclic AMP (cAMP) and is comprised of four subtypes (PDE4A–D). Previous studies have shown the beneficial effects of pan-PDE4 inhibitors in rodent PH;however, this class of drugs is associated with side effects owing to the broad inhibition of all four PDE4 isozymes. Here, we demonstrate that PDE4B is the predominant PDE isozyme in lungs and that it was upregulated in rodent and human PH lung tissues. We also confirmed that PDE4B is mainly expressed in the lung endothelial cells (ECs). Evaluation of PH in Pde4b wild type and knockout mice confirmed that Pde4b is important for the vascular remodeling associated with PH. In vivo EC lineage tracing demonstrated that Pde4b induces PH development by driving endothelial-to-mesenchymal transition (EndMT), and mechanistic studies showed that Pde4b regulates EndMT by antagonizing the cAMP-dependent PKA–CREB–BMPRII axis. Finally, treating PH rats with a PDE4B-specific inhibitor validated that PDE4B inhibition has a significant pharmacological effect in the alleviation of PH. Collectively, our findings indicate a critical role for PDE4B in EndMT and PH, prompting further studies of PDE4B-specific inhibitors as a therapeutic strategy for PH.
基金supported by the National Key R&D Program of China(2021YFD2100103)the National Natural Science Foundation of China(32101935).
文摘Glycidol is a common lipid-derived foodborne toxicant mainly presents in refined oils and related foodstuffs.Vascular endothelial cells may be potential targets of the deleterious effects associated with glycidol exposure.In human umbilical vein endothelial cells(HUVECs),we found that glycidol treatment promoted endothelialto-mesenchymal transition(EndMT)at a lower concentration(0.5 mmol/L),while induced apoptosis and inflammation at a higher concentration(1 mmol/L).These harmful effects were achieved by the activation of NF-κB/MAPK signaling pathway and were mediated by reactive oxygen species(ROS).In addition,the protective potential of 6-C-(E-2-fluorostyryl)naringenin(6-CEFN)against glycidol was evaluated and compared with naringenin.HUVECs pre-treated with 6-CEFN,but not naringenin,displayed resistance to endothelial dysfunction caused by glycidol.
基金Supported by the Scientific Research Project of the Traditional Chinese Medicine Bureau of Guangdong Province,China(Nos.20211136 and 20221086)。
文摘Objective:To investigate whether Leech-Centipede(LC) Granules can improve erectile function in rats with diabetes mellitus-associated erectile dysfunction(DMED) through endothelial-to-mesenchymal transition(EndMT) inhibition.Methods:Components of LC Granules were identified via ultra-high-performance liquid chromatography.Thirty male Sprague Dawley rats were injected with streptozotocin and fed continuously for 8weeks to establish the DMED rat model.Rats with erectile dysfunction symptoms diagnosed using apomorphine were divided into DMED and low-,medium-,and high-doses LC groups(n=6 in each).The negative control(NC, n=6) and DMED groups were given 5 mL of deionized water via intragastric gavage,and the low-,mediumand the high-doses LC groups were administered LC at 1.6,3.2,and 6.4 g/kg,respectively,via intragastric gavage for 4 weeks.The intracavernous pressure(ICP),mean arterial pressure(MAP),and nitric oxide(NO) levels in cavernous tissue were measured for each group.Quantitative reverse transcription-polymerase chain reaction and Western blot were used to detect mRNA and protein expressions of endothelial and mesenchymal markers.Immunofluorescence staining was used to observe α-SMA, and Masson’s trichrome staining was performed to determine the myofiber/collagen ratio.Results:A total of 474 active components were identified.After treatment,the ICP/MAP value and NO level were significantly higher in the medium-and high-dose LC groups than in the DMED group(P<0.05).Compared with the DMED groups,the medium-and high-dose groups LC significantly increased and decreased endothelial and mesenchymal markers expression,respectively(P<0.05).Tumor growth factor(TGF) β RⅡ,p-Smad2, and p-Smad3 levels were considerably higher following diabetes onset but reduced following LC intervention(P<0.05),except for TGF β 1(P>0.05).α-SMA expression was significantly higher in the DMED group and was reduced in all LC intervention groups(P>0.05).The myofiber/collagen ratio in the LC groups was higher than that in the DMED group but lower than that in the NC group(all P<0.05).Conclusions:LC Granules may improve the erectile function of DMED rats by suppressing TGF-β/Smad pathway to reverse EndMT.
基金supported by the Key Projects of the National Natural Science Foundation of China(81830055)National Natural Science Foundation of China(82001966)+1 种基金Outstanding Scientist Project of Chongqing(cstc2022ycjh-bgzxm0186)Special Funding for Postdoctoral Research of Chongqing(2021XM1020).
文摘Small-diameter tissue-engineered vascular grafts(sdTEVGs)have garnered significant attention as a potential treatment modality for vascular bypass grafting and replacement therapy.However,the intimal hyperplasia and thrombosis are two major complications that impair graft patency during transplantation.To address this issue,we fabricated the covalent-organic framework(COF)-based carbon monoxide(CO)nanogenerator-and co-immobilized with LXW-7 peptide and heparin to establish a multifunctional surface on TEVGs constructed from acellular blood vessels for preventing thrombosis and stenosis.The cell-adhesive peptide LXW-7 could capture endothelial-forming cells(EFCs)to promote endothelialization,while the antithrombotic molecule heparin prevented thrombus formation.The reactive oxygen species(ROS)-triggered CO release suppressed the adhesion and activation of macrophages,leading to the reduction of ROS and inflammatory factors.As a result,the endothelial-to-mesenchymal transition(EndMT)triggered by inflammation was restricted,facilitating the maintenance of the homeostasis of the neo-endothelium and preventing pathological remodeling in TEVGs.When transplanted in vivo,these vascular grafts exhibited negligible intimal hyperplasia and remained patent for 3 months.This achievement provided a novel approach for constructing antithrombotic and anti-hyperplastic TEVGs.
