目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过...目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过表达组、miR-451a敲低组、温肺降浊方含药血清(WFJZF)组、miR-451a过表达+WFJZF组、miR-451a敲低+WFJZF组和miR-451a空载体组,双荧光素酶确定miR-451a和PGAM5的转录关系,免疫蛋白共沉淀确定PGAM5和Drp1的调控作用,慢病毒转染miR-451a于HT22细胞中,qRT-PCR确定转染效率,CCK-8测定缺糖缺氧状态下细胞活性的最佳时间点,qRT-PCR和Westem Blot检测HT22模型细胞内DGAM5-Drp1轴蛋白及mRNA的表达水平。结果:OGD/R诱导HT22细胞持续2 h为最佳时间点,miR-451a可以靶向调控PGAM5基因,PGAM5和Drp1具有相互调控作用。与正常对照组比较,OGD/R组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达上升,p-Drp1 Ser 616磷酸化表达上升(P<0.05),OPA1蛋白及mRNA表达下降,p-Drp1 Ser 637去磷酸化表达下降(P<0.05);与OGD/R组比较,miR-451a过表达+WFJZF组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达下降,p-Drp1 Ser 616磷酸化表达下降(P<0.05),OPA1蛋白及mRNA表达上升,p-Drp1 Ser 637去磷酸化表达上升(P<0.05)。结论:miR-451a可以靶向调控PGAM5-Drp1轴,miR-451a过表达+WFJZF组可以改善线粒体失衡状态,减少神经元过度损伤,发挥脑保护效应。展开更多
Background Poor feather growth not only affects the appearance of the organism but also decreases the feed efficiency.Methionine(Met)is an essential amino acid required for feather follicle development;yet the exact m...Background Poor feather growth not only affects the appearance of the organism but also decreases the feed efficiency.Methionine(Met)is an essential amino acid required for feather follicle development;yet the exact mechanism involved remains insufficiently understood.Methods A total of 1801-day-old broilers were selected and randomly divided into 3 treatments:control group(0.45%Met),Met-deficiency group(0.25%Met),and Met-rescue group(0.45%Met in the pre-trial period and 0.25%Met in the post-trial period).The experimental period lasted for 56 d,with a pre-trial period of 1–28 d and a post-trial period of 29–56 d.In addition,Met-deficiency and Met-rescue models were constructed in feather follicle epidermal stem cell by controlling the supply of Met in the culture medium.Results Dietary Met-deficiency significantly(P<0.05)reduced the ADG,ADFI and F/G,and inhibited feather follicle development.Met supplementation significantly(P<0.05)improved growth performance and the feather growth in broilers.Met-rescue may promote feather growth in broilers by activating the Wnt/β-catenin signaling pathway(GSK-3β,CK1,Axin1,β-catenin,Activeβ-catenin,TCF4,and Cyclin D1).Compared with Met-deficiency group,Met-rescue significantly(P<0.05)increased the activity of feather follicle epidermal stem cell and mitochondrial membrane potential,activated Wnt/β-catenin signaling pathway,and decreased the content of reactive oxygen species(P<0.05).CO-IP confirmed that mitochondrial protein PGAM5 interacted with Axin1,the scaffold protein of the disruption complex of the Wnt/β-catenin signaling pathway,and directly mediated Met regulation of Wnt/β-catenin signaling pathway and feather follicle development.Conclusions PGAM5 binding to Axin1 mediates the regulation of Wnt/β-catenin signaling pathway,and promotes feather follicle development and feather growth of broiler chickens through Met supplementation.These results provide theoretical support for the improvement of economic value and production efficiency of broiler chickens.展开更多
目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁...目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁正常组织(P<0.05)。结论:PGAM1在乳腺癌组织中有高表达,为乳腺癌的诊断及治疗提供了新的思路。展开更多
文摘目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过表达组、miR-451a敲低组、温肺降浊方含药血清(WFJZF)组、miR-451a过表达+WFJZF组、miR-451a敲低+WFJZF组和miR-451a空载体组,双荧光素酶确定miR-451a和PGAM5的转录关系,免疫蛋白共沉淀确定PGAM5和Drp1的调控作用,慢病毒转染miR-451a于HT22细胞中,qRT-PCR确定转染效率,CCK-8测定缺糖缺氧状态下细胞活性的最佳时间点,qRT-PCR和Westem Blot检测HT22模型细胞内DGAM5-Drp1轴蛋白及mRNA的表达水平。结果:OGD/R诱导HT22细胞持续2 h为最佳时间点,miR-451a可以靶向调控PGAM5基因,PGAM5和Drp1具有相互调控作用。与正常对照组比较,OGD/R组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达上升,p-Drp1 Ser 616磷酸化表达上升(P<0.05),OPA1蛋白及mRNA表达下降,p-Drp1 Ser 637去磷酸化表达下降(P<0.05);与OGD/R组比较,miR-451a过表达+WFJZF组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达下降,p-Drp1 Ser 616磷酸化表达下降(P<0.05),OPA1蛋白及mRNA表达上升,p-Drp1 Ser 637去磷酸化表达上升(P<0.05)。结论:miR-451a可以靶向调控PGAM5-Drp1轴,miR-451a过表达+WFJZF组可以改善线粒体失衡状态,减少神经元过度损伤,发挥脑保护效应。
基金supported by the National Natural Science Foundation of China(32372902)Guangdong Special Support Program for Young Talents(NYQN2024007)。
文摘Background Poor feather growth not only affects the appearance of the organism but also decreases the feed efficiency.Methionine(Met)is an essential amino acid required for feather follicle development;yet the exact mechanism involved remains insufficiently understood.Methods A total of 1801-day-old broilers were selected and randomly divided into 3 treatments:control group(0.45%Met),Met-deficiency group(0.25%Met),and Met-rescue group(0.45%Met in the pre-trial period and 0.25%Met in the post-trial period).The experimental period lasted for 56 d,with a pre-trial period of 1–28 d and a post-trial period of 29–56 d.In addition,Met-deficiency and Met-rescue models were constructed in feather follicle epidermal stem cell by controlling the supply of Met in the culture medium.Results Dietary Met-deficiency significantly(P<0.05)reduced the ADG,ADFI and F/G,and inhibited feather follicle development.Met supplementation significantly(P<0.05)improved growth performance and the feather growth in broilers.Met-rescue may promote feather growth in broilers by activating the Wnt/β-catenin signaling pathway(GSK-3β,CK1,Axin1,β-catenin,Activeβ-catenin,TCF4,and Cyclin D1).Compared with Met-deficiency group,Met-rescue significantly(P<0.05)increased the activity of feather follicle epidermal stem cell and mitochondrial membrane potential,activated Wnt/β-catenin signaling pathway,and decreased the content of reactive oxygen species(P<0.05).CO-IP confirmed that mitochondrial protein PGAM5 interacted with Axin1,the scaffold protein of the disruption complex of the Wnt/β-catenin signaling pathway,and directly mediated Met regulation of Wnt/β-catenin signaling pathway and feather follicle development.Conclusions PGAM5 binding to Axin1 mediates the regulation of Wnt/β-catenin signaling pathway,and promotes feather follicle development and feather growth of broiler chickens through Met supplementation.These results provide theoretical support for the improvement of economic value and production efficiency of broiler chickens.
文摘目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁正常组织(P<0.05)。结论:PGAM1在乳腺癌组织中有高表达,为乳腺癌的诊断及治疗提供了新的思路。
