AIM:To investigate the effects of Sonic hedgehog(Shh)gene-modified bone marrow mesenchymal stem cells(MSCs)on graft-induced retinal gliosis and retinal ganglion cells(RGCs)survival in diabetic mice.METHODS:Bone marrow...AIM:To investigate the effects of Sonic hedgehog(Shh)gene-modified bone marrow mesenchymal stem cells(MSCs)on graft-induced retinal gliosis and retinal ganglion cells(RGCs)survival in diabetic mice.METHODS:Bone marrow-derived MSCs were genetically modified with the Shh gene to generate a stably transfected cell line of Shh-modified MSCs(MSC-Shh).Intravitreal injections of MSC-Shh and green fluorescent protein-modified MSCs(MSC-Gfp;control)were administered in diabetic mice.After 4wk,the effects of MSC-Shh on retinal gliosis were evaluated using fundus photography,and markers of gliosis were examined by immunofluorescence and Western blotting.The neurotrophic factors expression and RGCs survival in the host retina were evaluated using Western blotting and immunofluorescence.The mechanisms underlying the effects of MSC-Shh was investigated.RESULTS:A significant reduction of proliferative vitreoretinopathy(PVR)was observed after intravitreal injection of MSC-Shh compared to MSC-Gfp.Significant downregulation of glial fibrillary acidic protein(GFAP)was demonstrated in the host retina after MSC-Shh administration compared to MSC-Gfp.The extracellular signal-regulated kinase 1/2(ERK1/2),protein kinase B(AKT)and phosphatidylin-ositol-3-kinase(PI3K)pathways were significantly downregulated after MSC-Shh administration compared to MSC-Gfp.Brain-derived neurotrophic factor(BDNF)and ciliary neurotrophic factor(CNTF)levels were significantly increased in the host retina,and RGCs loss was significantly prevented after MSC-Shh administration.CONCLUSION:MSC-Shh administration reduces graft-induced reactive gliosis following intravitreal injection in diabetic mice.The ERK1/2,AKT and PI3K pathways are involved in this process.MSC-Shh also increases the levels of neurotrophic factors in the host retina and promoted RGCs survival in diabetic mice.展开更多
Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embry...Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.展开更多
目的:研究大鼠胃癌前病变形成过程中胃黏膜Sonic Hedgehog(Shh)信号通路的变化。方法:72只乳鼠随机分为正常组36只,造模组36只,雌雄各半。造模组灌服800 mg/L的1-甲基-3-硝基-1-亚硝基胍(MNNG)0.1 m L/d,正常组灌服生理盐水0.1 m L/d,共...目的:研究大鼠胃癌前病变形成过程中胃黏膜Sonic Hedgehog(Shh)信号通路的变化。方法:72只乳鼠随机分为正常组36只,造模组36只,雌雄各半。造模组灌服800 mg/L的1-甲基-3-硝基-1-亚硝基胍(MNNG)0.1 m L/d,正常组灌服生理盐水0.1 m L/d,共10 d。随后在实验观察第10周、22周、34周末每组分别雌雄各处死6只。HE染色检测胃黏膜组织病理学变化,q RTPCR法检测胃黏膜组织Shh、Ptch1、Smo、Gli1、Gli2、Gli3、Su Fu、Cyclin D1、Cyclin E1、c-Myc、β-actin m RNA的表达。Western blot检测胃黏膜组织Shh、Ptch1、Smo、Gli1、Su Fu、Cyclin D1、Cyclin E1、cMyc、p-c-Myc蛋白表达。结果:随造模时间的延长,模型组大鼠胃黏膜萎缩及异型增生逐渐加重,其胃黏膜内Shh、Smo、Gli1、Cyclin D1、Cyclin E1及c-Myc m RNA表达较正常组不同程度升高,而Ptch1和Su Fu m RNA不同程度降低。模型组胃黏膜内Shh、Smo、Gli1、Cyclin D1、Cyclin E1、p-c-Myc蛋白水平较正常组增加,Ptch1和Su Fu则降低,但差异无统计学意义。结论:大鼠胃癌前病变的形成过程中存在Shh信号通路的激活,提示Shh信号通路可能参与胃癌前病变的发生与发展。展开更多
基金Supported by the Natural Science Foundation of Guangdong Province(No.2018A0303130293,No.2023A1515012470).
文摘AIM:To investigate the effects of Sonic hedgehog(Shh)gene-modified bone marrow mesenchymal stem cells(MSCs)on graft-induced retinal gliosis and retinal ganglion cells(RGCs)survival in diabetic mice.METHODS:Bone marrow-derived MSCs were genetically modified with the Shh gene to generate a stably transfected cell line of Shh-modified MSCs(MSC-Shh).Intravitreal injections of MSC-Shh and green fluorescent protein-modified MSCs(MSC-Gfp;control)were administered in diabetic mice.After 4wk,the effects of MSC-Shh on retinal gliosis were evaluated using fundus photography,and markers of gliosis were examined by immunofluorescence and Western blotting.The neurotrophic factors expression and RGCs survival in the host retina were evaluated using Western blotting and immunofluorescence.The mechanisms underlying the effects of MSC-Shh was investigated.RESULTS:A significant reduction of proliferative vitreoretinopathy(PVR)was observed after intravitreal injection of MSC-Shh compared to MSC-Gfp.Significant downregulation of glial fibrillary acidic protein(GFAP)was demonstrated in the host retina after MSC-Shh administration compared to MSC-Gfp.The extracellular signal-regulated kinase 1/2(ERK1/2),protein kinase B(AKT)and phosphatidylin-ositol-3-kinase(PI3K)pathways were significantly downregulated after MSC-Shh administration compared to MSC-Gfp.Brain-derived neurotrophic factor(BDNF)and ciliary neurotrophic factor(CNTF)levels were significantly increased in the host retina,and RGCs loss was significantly prevented after MSC-Shh administration.CONCLUSION:MSC-Shh administration reduces graft-induced reactive gliosis following intravitreal injection in diabetic mice.The ERK1/2,AKT and PI3K pathways are involved in this process.MSC-Shh also increases the levels of neurotrophic factors in the host retina and promoted RGCs survival in diabetic mice.
基金supported by the Ministry of EducationScience and Technology(No.2021M3A9A1096894)+1 种基金Republic of Korea and the KRIBB Research Initiative Program(KGM4252223)Korea Research Institute of Bioscience and Biotechnology,Republic of Korea。
文摘Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.
文摘目的:研究大鼠胃癌前病变形成过程中胃黏膜Sonic Hedgehog(Shh)信号通路的变化。方法:72只乳鼠随机分为正常组36只,造模组36只,雌雄各半。造模组灌服800 mg/L的1-甲基-3-硝基-1-亚硝基胍(MNNG)0.1 m L/d,正常组灌服生理盐水0.1 m L/d,共10 d。随后在实验观察第10周、22周、34周末每组分别雌雄各处死6只。HE染色检测胃黏膜组织病理学变化,q RTPCR法检测胃黏膜组织Shh、Ptch1、Smo、Gli1、Gli2、Gli3、Su Fu、Cyclin D1、Cyclin E1、c-Myc、β-actin m RNA的表达。Western blot检测胃黏膜组织Shh、Ptch1、Smo、Gli1、Su Fu、Cyclin D1、Cyclin E1、cMyc、p-c-Myc蛋白表达。结果:随造模时间的延长,模型组大鼠胃黏膜萎缩及异型增生逐渐加重,其胃黏膜内Shh、Smo、Gli1、Cyclin D1、Cyclin E1及c-Myc m RNA表达较正常组不同程度升高,而Ptch1和Su Fu m RNA不同程度降低。模型组胃黏膜内Shh、Smo、Gli1、Cyclin D1、Cyclin E1、p-c-Myc蛋白水平较正常组增加,Ptch1和Su Fu则降低,但差异无统计学意义。结论:大鼠胃癌前病变的形成过程中存在Shh信号通路的激活,提示Shh信号通路可能参与胃癌前病变的发生与发展。
