背景:Sema7A是一种细胞表面蛋白,它可以促进破骨细胞的融合同时促进成骨细胞的迁移,从而影响骨的动态平衡。推测,Sema7A siR NA可能减弱钛微粒成骨细胞分化。目的:分析信号素7A(Sema7A)对钛颗粒抑制鼠MC3T3-E1前成骨细胞活性过程的影响...背景:Sema7A是一种细胞表面蛋白,它可以促进破骨细胞的融合同时促进成骨细胞的迁移,从而影响骨的动态平衡。推测,Sema7A siR NA可能减弱钛微粒成骨细胞分化。目的:分析信号素7A(Sema7A)对钛颗粒抑制鼠MC3T3-E1前成骨细胞活性过程的影响。方法:取第六七代生长状态良好的小鼠颅骨前成骨细胞(MC3T3-E1),分为4组培养。其中空白对照组为单独培养的细胞;标准对照组加入了钛微粒共培养;实验1组加入Sema7A过表达质粒,实验2组加入Sema7AsiR NA进行干扰。采用流式细胞仪检测细胞凋亡情况;Q-PCR检测骨涎蛋白、骨钙素、Ⅰ型胶原蛋白的mR NA表达情况;Western-blot检测骨涎蛋白、骨钙素、Ⅰ型胶原蛋白的蛋白表达情况;茜素红钙结节染色检测成骨细胞的矿化程度。结果与结论:结果提示标准对照组、实验1组检测到骨涎蛋白、骨钙素、Ⅰ型胶原蛋白表达明显减少而实验2组骨涎蛋白、骨钙素、Ⅰ型胶原蛋白则比标准对照组表达增加(P<0.05)。流式细胞仪结果提示实验1组成骨细胞凋亡率明显高于其他组(P<0.05),而实验2组则比标准对照组凋亡率降低(P<0.05)。茜素红染色检测结果显示实验1组未见明显的矿化结节,实验2组有矿化结节形成。结果表明通过基因干扰技术抑制Sema7A活性可以对钛颗粒抑制鼠MC3T3-E1成骨细胞分化的过程进行干扰,进而为临床上用生物技术治疗磨损微粒诱导的骨溶解问题提供了一条可行的思路。展开更多
Background:Vascular smooth muscle cells(VSMCs)undergo a conversion from a contractile phenotype to a proliferative synthetic phenotype,contributing to the pathogenesis of cardiovascular diseases.Semaphorin 7A(SEMA7A)i...Background:Vascular smooth muscle cells(VSMCs)undergo a conversion from a contractile phenotype to a proliferative synthetic phenotype,contributing to the pathogenesis of cardiovascular diseases.Semaphorin 7A(SEMA7A)is a glycosylphosphatidylinositol-anchored membrane protein that plays an important role in vascular homeostasis by regulating endothelial cell behaviors.However,the expression and role of SEMA7A in VSMCs remain unclear.Methods:In this study,we screened for VSMC-regulating genes in publicly available datasets and analyzed the expression of SEMA7A in human coronary artery smooth muscle cells(hCASMCs)treated with platelet-derived growth factor-BB(PDGF-BB).The effects of SEMA7A overexpression and knockdown on hCASMC proliferation and migration were examined.The signaling pathways involved in the action of SEMA7A in hCASMCs were determined.Results:Bioinformatic analysis showed that SEMA7A was significantly dysregulated in VSMCs treated with oxidized low-density lipoprotein or overexpressing progerin,a pro-atherogenic gene.The PDGF-BB stimulation led to a concentration-and time-dependent induction of SEMA7A.Depletion of SEMA7A attenuated PDGF-BB-induced hCASMC proliferation and migration.Conversely,overexpression of SEMA7A enhanced hCASMC proliferation and migration.Mechanistically,SEMA7A stimulated the activation of theβ-catenin pathway and upregulated c-Myc,CCND1,and MMP7.Knockdown ofβ-catenin impaired SEMA7A-induced hCASMC proliferation and migration.Conclusions:SEMA7A triggers phenotype switching in VSMCs through theβ-catenin signaling pathway and may serve as a potential therapeutic target for cardiovascular diseases.展开更多
Atherosclerosis is a chronic inflammatory disease driven by pathological processes such as macrophage foam cell formation.Semaphorin 7A(SEMA7A)is an immunoregulatory signaling molecule known to modulate immune respons...Atherosclerosis is a chronic inflammatory disease driven by pathological processes such as macrophage foam cell formation.Semaphorin 7A(SEMA7A)is an immunoregulatory signaling molecule known to modulate immune responses and cellular adhesion.However,the contribution of macrophage-derived SEMA7A to atherogenesis has yet to be fully elucidated.In this study,we analyzed gene expression profiles of human mononuclear cells from the Gene Expression Omnibus(GEO)database and revealed highly expressed SEMA7A and its receptor integrin β1 in macrophages.The upregulation of SEMA7A and integrin β1 was also observed during the differentiation of THP-1 monocytes into macrophages.Mice with macrophage-specific deletion of Sema7a showed a 57.2%reduction in atherosclerotic lesion size and improved plaque stability in atherosclerosis mouse model compared to control mice.Mechanistically,macrophage SEMA7A promoted the expression of macrophage scavenger receptor 1(MSR1)and lipid uptake mediated by integrin β1 and downstream JNK signaling pathway in macrophages.Notably,pharmacological inhibition of integrin β1 with integrin receptor antagonist GLPG0187 effectively suppressed atherosclerosis progression.These findings identify macrophage-derived SEMA7A as a key driver of atherosclerosis through a novel integrin β1/JNK/MSR1 axis,providing potential targets for the prevention and treatment of atherosclerosis.展开更多
文摘背景:Sema7A是一种细胞表面蛋白,它可以促进破骨细胞的融合同时促进成骨细胞的迁移,从而影响骨的动态平衡。推测,Sema7A siR NA可能减弱钛微粒成骨细胞分化。目的:分析信号素7A(Sema7A)对钛颗粒抑制鼠MC3T3-E1前成骨细胞活性过程的影响。方法:取第六七代生长状态良好的小鼠颅骨前成骨细胞(MC3T3-E1),分为4组培养。其中空白对照组为单独培养的细胞;标准对照组加入了钛微粒共培养;实验1组加入Sema7A过表达质粒,实验2组加入Sema7AsiR NA进行干扰。采用流式细胞仪检测细胞凋亡情况;Q-PCR检测骨涎蛋白、骨钙素、Ⅰ型胶原蛋白的mR NA表达情况;Western-blot检测骨涎蛋白、骨钙素、Ⅰ型胶原蛋白的蛋白表达情况;茜素红钙结节染色检测成骨细胞的矿化程度。结果与结论:结果提示标准对照组、实验1组检测到骨涎蛋白、骨钙素、Ⅰ型胶原蛋白表达明显减少而实验2组骨涎蛋白、骨钙素、Ⅰ型胶原蛋白则比标准对照组表达增加(P<0.05)。流式细胞仪结果提示实验1组成骨细胞凋亡率明显高于其他组(P<0.05),而实验2组则比标准对照组凋亡率降低(P<0.05)。茜素红染色检测结果显示实验1组未见明显的矿化结节,实验2组有矿化结节形成。结果表明通过基因干扰技术抑制Sema7A活性可以对钛颗粒抑制鼠MC3T3-E1成骨细胞分化的过程进行干扰,进而为临床上用生物技术治疗磨损微粒诱导的骨溶解问题提供了一条可行的思路。
基金supported by the Basic Research Program of Shanxi Province(Free Exploration)of China(20210302124416)Science and Technology Grant for Selected Returned Chinese Scholars of Shanxi Province of China(20220043)Four“Batches”Innovation Project of Invigorating Medical through Science and Technology of Shanxi Province of China(2022XM08).
文摘Background:Vascular smooth muscle cells(VSMCs)undergo a conversion from a contractile phenotype to a proliferative synthetic phenotype,contributing to the pathogenesis of cardiovascular diseases.Semaphorin 7A(SEMA7A)is a glycosylphosphatidylinositol-anchored membrane protein that plays an important role in vascular homeostasis by regulating endothelial cell behaviors.However,the expression and role of SEMA7A in VSMCs remain unclear.Methods:In this study,we screened for VSMC-regulating genes in publicly available datasets and analyzed the expression of SEMA7A in human coronary artery smooth muscle cells(hCASMCs)treated with platelet-derived growth factor-BB(PDGF-BB).The effects of SEMA7A overexpression and knockdown on hCASMC proliferation and migration were examined.The signaling pathways involved in the action of SEMA7A in hCASMCs were determined.Results:Bioinformatic analysis showed that SEMA7A was significantly dysregulated in VSMCs treated with oxidized low-density lipoprotein or overexpressing progerin,a pro-atherogenic gene.The PDGF-BB stimulation led to a concentration-and time-dependent induction of SEMA7A.Depletion of SEMA7A attenuated PDGF-BB-induced hCASMC proliferation and migration.Conversely,overexpression of SEMA7A enhanced hCASMC proliferation and migration.Mechanistically,SEMA7A stimulated the activation of theβ-catenin pathway and upregulated c-Myc,CCND1,and MMP7.Knockdown ofβ-catenin impaired SEMA7A-induced hCASMC proliferation and migration.Conclusions:SEMA7A triggers phenotype switching in VSMCs through theβ-catenin signaling pathway and may serve as a potential therapeutic target for cardiovascular diseases.
基金supported by the National Natural Science Foundation of China(Nos.82170466,82470483 to Li Zhu,82070450 to Chaojun Tang)the Natural Science Foundation of Jiangsu Province(No.BK20230487 to Fengchan Li)+1 种基金the fellowship of China Postdoctoral Science Foundation(No.7121102223 to Fengchan Li)the Priority Academic Program Development of Jiangsu Higher Education Institutions of China(to Li Zhu).
文摘Atherosclerosis is a chronic inflammatory disease driven by pathological processes such as macrophage foam cell formation.Semaphorin 7A(SEMA7A)is an immunoregulatory signaling molecule known to modulate immune responses and cellular adhesion.However,the contribution of macrophage-derived SEMA7A to atherogenesis has yet to be fully elucidated.In this study,we analyzed gene expression profiles of human mononuclear cells from the Gene Expression Omnibus(GEO)database and revealed highly expressed SEMA7A and its receptor integrin β1 in macrophages.The upregulation of SEMA7A and integrin β1 was also observed during the differentiation of THP-1 monocytes into macrophages.Mice with macrophage-specific deletion of Sema7a showed a 57.2%reduction in atherosclerotic lesion size and improved plaque stability in atherosclerosis mouse model compared to control mice.Mechanistically,macrophage SEMA7A promoted the expression of macrophage scavenger receptor 1(MSR1)and lipid uptake mediated by integrin β1 and downstream JNK signaling pathway in macrophages.Notably,pharmacological inhibition of integrin β1 with integrin receptor antagonist GLPG0187 effectively suppressed atherosclerosis progression.These findings identify macrophage-derived SEMA7A as a key driver of atherosclerosis through a novel integrin β1/JNK/MSR1 axis,providing potential targets for the prevention and treatment of atherosclerosis.
