HIF-1α和Hsp90抑制剂对福美双诱导的胫骨软骨发育不良肉鸡生长板功能恢复的影响
发布时间：2019-05-26 21:48
【摘要】：肉鸡胫骨软骨发育不良(Tibial dyschondroplasia,TD)是一种以生长板无血管化和无矿化为特征的重要家禽胫跗骨疾病。它主要引起软骨细胞的异常分化和家禽的跛行,从而降低动物的生长性能,对家禽业造成巨大的经济损失。1.HIF-1ɑ抑制剂在福美双诱导的TD上的作用本研究旨在评估缺氧诱导因子-1α(Hypoxia-Inducible Factor-1α,HIF-1ɑ)抑制剂FK228在福美双诱导的TD上的作用和HIF-1ɑ和Hsp90之间的关系。将150只肉鸡随机分成3组:对照组、福美双组、FK228组。通过RT-qPCR检测第10天和14天HIF-1ɑ、Hsp90和血管内皮生长因子(vascular endothelial growth factor,VEGF)基因的表达;Western blot检测第14天HIF-1ɑ和Hsp90蛋白表达水平。结果显示:在TD病程中HIF-1ɑ显著上调(P0.05),VEGF没有显示变化(p0.05)。HIF-1ɑ抑制剂能够显著下调HIF-1ɑ和VEGF基因的表达和蛋白的表达水平(P0.05),从而减少生长板的尺寸和肉鸡的跛行。综上所述,HIF-1ɑ和VEGF在肉鸡胫骨软骨发育不良非血管化的生长板上在形成中起着重要作用,这可能就是作为防治肉鸡无血管化生长板和跛行的HIF-1ɑ抑制因子作用的靶点。HIF-1ɑ抑制因子可以作为预防肉鸡胫骨软骨生长板无血管化、生长跛行和恢复生长板血管生成的新靶标。2.Hsp90抑制剂EGCG和芹黄素在肉鸡胫骨软骨发育不良上具有治愈生长板的作用热休克蛋白90(Heat-shock protein 90,Hsp90)在动物组织中是一种前血管生长因子,研究表明在肉鸡TD的病例中Hsp90表达量增加。本研究旨在评估Hsp90抑制剂表没食子儿茶素没食子酸酯(EGCG)和芹黄素在福美双诱导的肉鸡TD中的作用。本研究采用HE对生长板进行组织学观察,RT-qPCR检测Hsp90基因的表达量。结果表明:肉鸡TD的发生伴随血管减少,软骨细胞分化发生改变,Hsp90表达量显著增加(P0.05)。然而,TD肉鸡给予EGCG和芹黄素后发现软骨细胞柱状组织修复和Hsp90表达活性被降低(P0.05),最终跛行消失。结果表明Hsp90是TD发生的一个关键因素;EGCG和芹黄素通过抑制Hsp90基因的表达来有效的治疗TD,减少跛行和肉鸡腿的畸形。3.HIF-1α抑制剂Apigenin在生长板软骨治愈上的作用本研究旨在了解缺氧诱导因子(HIF-1α)抑制剂的机制和芹黄素在福美双诱导TD上的抗氧化能力。将150只肉鸡随机分为3组:对照组、福美双组、芹黄素组。分别于试验的第10天和第14天采用RT-qPCR方法对HIF-1ɑ基因表达量进行检测。结果表明:与对照组相比,HIF-1ɑ基因表达量在福美双诱发TD期间显著上调(P0.05);芹黄素组HIF-1ɑ表达量显著下调(P0.05),生长板尺寸和跛行减少。另外,福美双诱导肝脏中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)和丙二醛(MDA)含量的增加,而福美双组抗氧化酶(SOD;GSH-Px)和碱性磷酸酶(ALP)值降低;而芹黄素组各项指标均接近正常组。结果表明:HIF-1ɑ抑制剂能够恢复肉鸡跛行和非血管化的生长板,且芹黄素能够有效的改善福美双引起的肝脏损伤和氧化失衡。
[Abstract]:Dystrophy of tibia cartilage (Tibial dyschondroplasia,TD) in broilers is an important disease of tibia and tarsal bone in poultry characterized by non-vascularization and mineralization of growth plate. It mainly causes the abnormal differentiation of cartilage cells and the claudication of poultry, thus reducing the growth performance of animals. The effect of HIF-1 inhibitor on TD induced by Fumei Shuang was studied in order to evaluate hypoxia-inducible factor-1 伪 (Hypoxia-Inducible Factor-1 伪). 1. The effect of HIF-1 inhibitor on Fumei-induced HIF-1 伪 was studied in order to evaluate hypoxia-induced factor-1 伪 (HIF-伪). The role of FK228, an inhibitor of HIF-1, in TD induced by Fumibi and the relationship between HIF-1 and Hsp90. 150 broilers were randomly divided into three groups: control group, Fumei group and FK228 group. The expression of HIF-1, Hsp90 and vascular endothelial growth factor (vascular endothelial growth factor,VEGF (vascular endothelial growth factor,VEGF) genes on the 10th and 14th day were detected by RT-qPCR. The expression levels of HIF-1 and Hsp90 proteins on the 14th day were detected by; Western blot. The results showed that HIF-1 was significantly up-regulated in the course of TD (p0.05). HIF-1 inhibitor could significantly down-regulate the expression of HIF-1 and VEGF genes and the expression of protein (P 0.05). Thus reducing the size of the growth board and the limp of broilers. To sum up, HIF-1 and VEGF play an important role in the formation of non-vascularized growth plates with dysplasia of tibia in broilers. This may be the target for the prevention and treatment of avascularization growth plate and claudication inhibitor of HIF-1 in broilers. HIF-1 inhibitor can be used to prevent the absence of vascularization of tibia cartilage growth plate in broilers. A new target for growth claudication and recovery of growth plate vascularization. 2. Hsp90 inhibitor EGCG and celery can cure growth plate heat shock protein 90 (Heat-shock protein 90) in broilers with dystrophy of tibia cartilage. Hsp90) is a pre-vascular growth factor in animal tissues. Studies have shown that the expression of Hsp90 is increased in broilers with TD. The purpose of this study was to evaluate the role of Hsp90 inhibitors epigallocatechin gallate (EGCG) and celery in TD induced by Fu Mei Shuang. In this study, HE was used to observe the histology of the growth plate, and the expression of Hsp90 gene was detected by RT-qPCR. The results showed that the occurrence of TD in broilers was accompanied by the decrease of blood vessels, the differentiation of cartilage cells and the increase of Hsp90 expression (P 0.05). However, after administration of EGCG and celery in TD broilers, it was found that the columnar tissue repair and Hsp90 expression activity of cartilage cells were decreased (P 0.05), and finally limp disappeared. The results showed that Hsp90 was a key factor in the occurrence of TD. EGCG and celery can effectively treat TD, by inhibiting the expression of Hsp90 gene. 3. The role of Apigenin, an inhibitor of HIF-1 伪, in the healing of growth plate cartilage. The purpose of this study was to investigate the mechanism of hypoxia inducible factor 伪 (HIF-1 伪) inhibitor and the antioxidant capacity of celery on Fumei induced TD. 150 broilers were randomly divided into three groups: control group, Fumei group and celery group. The expression of HIF-1 gene was detected by RT-qPCR on the 10th and 14th day of the experiment, respectively. The results showed that compared with the control group, the expression of HIF-1 gene was significantly up-regulated during TD induced by Fu Mei Shuang (P 0.05), while the expression of HIF-1 in celery group was significantly down-regulated (P 0.05), and the size of growth plate and claudication were decreased. In addition, the contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and malondialdehyde (MDA) in liver were increased, while the (ALP) values of antioxidant enzymes (SOD;GSH-Px) and alkaline phosphatase were decreased. However, all the indexes in celery group were close to those in normal group. The results showed that HIF-1 inhibitor could restore limp and non-vascularization growth plate in broilers, and celery could effectively improve liver injury and oxidative imbalance induced by Fu Mei Shuang.
【学位授予单位】：华中农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S858.31
本文编号：2485660
[Abstract]:Dystrophy of tibia cartilage (Tibial dyschondroplasia,TD) in broilers is an important disease of tibia and tarsal bone in poultry characterized by non-vascularization and mineralization of growth plate. It mainly causes the abnormal differentiation of cartilage cells and the claudication of poultry, thus reducing the growth performance of animals. The effect of HIF-1 inhibitor on TD induced by Fumei Shuang was studied in order to evaluate hypoxia-inducible factor-1 伪 (Hypoxia-Inducible Factor-1 伪). 1. The effect of HIF-1 inhibitor on Fumei-induced HIF-1 伪 was studied in order to evaluate hypoxia-induced factor-1 伪 (HIF-伪). The role of FK228, an inhibitor of HIF-1, in TD induced by Fumibi and the relationship between HIF-1 and Hsp90. 150 broilers were randomly divided into three groups: control group, Fumei group and FK228 group. The expression of HIF-1, Hsp90 and vascular endothelial growth factor (vascular endothelial growth factor,VEGF (vascular endothelial growth factor,VEGF) genes on the 10th and 14th day were detected by RT-qPCR. The expression levels of HIF-1 and Hsp90 proteins on the 14th day were detected by; Western blot. The results showed that HIF-1 was significantly up-regulated in the course of TD (p0.05). HIF-1 inhibitor could significantly down-regulate the expression of HIF-1 and VEGF genes and the expression of protein (P 0.05). Thus reducing the size of the growth board and the limp of broilers. To sum up, HIF-1 and VEGF play an important role in the formation of non-vascularized growth plates with dysplasia of tibia in broilers. This may be the target for the prevention and treatment of avascularization growth plate and claudication inhibitor of HIF-1 in broilers. HIF-1 inhibitor can be used to prevent the absence of vascularization of tibia cartilage growth plate in broilers. A new target for growth claudication and recovery of growth plate vascularization. 2. Hsp90 inhibitor EGCG and celery can cure growth plate heat shock protein 90 (Heat-shock protein 90) in broilers with dystrophy of tibia cartilage. Hsp90) is a pre-vascular growth factor in animal tissues. Studies have shown that the expression of Hsp90 is increased in broilers with TD. The purpose of this study was to evaluate the role of Hsp90 inhibitors epigallocatechin gallate (EGCG) and celery in TD induced by Fu Mei Shuang. In this study, HE was used to observe the histology of the growth plate, and the expression of Hsp90 gene was detected by RT-qPCR. The results showed that the occurrence of TD in broilers was accompanied by the decrease of blood vessels, the differentiation of cartilage cells and the increase of Hsp90 expression (P 0.05). However, after administration of EGCG and celery in TD broilers, it was found that the columnar tissue repair and Hsp90 expression activity of cartilage cells were decreased (P 0.05), and finally limp disappeared. The results showed that Hsp90 was a key factor in the occurrence of TD. EGCG and celery can effectively treat TD, by inhibiting the expression of Hsp90 gene. 3. The role of Apigenin, an inhibitor of HIF-1 伪, in the healing of growth plate cartilage. The purpose of this study was to investigate the mechanism of hypoxia inducible factor 伪 (HIF-1 伪) inhibitor and the antioxidant capacity of celery on Fumei induced TD. 150 broilers were randomly divided into three groups: control group, Fumei group and celery group. The expression of HIF-1 gene was detected by RT-qPCR on the 10th and 14th day of the experiment, respectively. The results showed that compared with the control group, the expression of HIF-1 gene was significantly up-regulated during TD induced by Fu Mei Shuang (P 0.05), while the expression of HIF-1 in celery group was significantly down-regulated (P 0.05), and the size of growth plate and claudication were decreased. In addition, the contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and malondialdehyde (MDA) in liver were increased, while the (ALP) values of antioxidant enzymes (SOD;GSH-Px) and alkaline phosphatase were decreased. However, all the indexes in celery group were close to those in normal group. The results showed that HIF-1 inhibitor could restore limp and non-vascularization growth plate in broilers, and celery could effectively improve liver injury and oxidative imbalance induced by Fu Mei Shuang.
【学位授予单位】：华中农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S858.31
【参考文献】
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1 Georgina N.Masoud;Wei Li;;HIF-1α pathway: role, regulation and intervention for cancer therapy[J];Acta Pharmaceutica Sinica B;2015年05期
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