不同产蛋期蛋鸡骨和蛋壳腺钙磷代谢相关因子表达变化的研究

发布时间：2018-04-17 10:28

  本文选题：钙磷代谢 + 骨　； 参考：《沈阳农业大学》2017年硕士论文

【摘要】：在现代集约化养殖体系中,蛋鸡骨质疏松症作为一种威胁笼养蛋鸡健康和生产性能的常见骨骼代谢疾病,给蛋鸡养殖业造成了严重损失。产蛋期蛋壳腺对钙的大量需求而引起骨形成和吸收失衡是蛋鸡骨质疏松发生的主要原因。本试验通过检测不同产蛋期蛋鸡胫骨形态学特性、骨和蛋壳腺中钙磷代谢相关因子以及蛋壳品质的变化,探讨骨和蛋壳腺中钙磷代谢相关因子变化与骨形态及蛋壳品质的相关性,以期为蛋鸡日粮中钙磷添加、骨质疏松的预防提供理论依据。试验随机选取产蛋前期(23周龄)、产蛋高峰期(43周龄)和产蛋后期(71周龄)的海兰褐蛋鸡作为试验对象,每时期各12只,采集血液并制备血清,比色法检测血清中Ca、P、ALP和TRAP含量;放射免疫分析法检测血清中PTH、CT、25-(OH)_2VD_3、1,25-(OH)_2VD_3、E_2和BGP含量；酶联免疫酶免法检测血清中PICP含量。采集右侧胫骨,通过micro-CT检测骨的微结构,骨切片TRAP染色观察破骨细胞变化。采集股骨关节和蛋壳腺,提取总RNA,通过实时荧光定量PCR法检测骨中钙磷代谢相关因子ALP、SPHK1、BMP2、BMP6、BMP7、PDGFB、TRAP、V-ATPasea3、RANKL、OPG、M-CSF、CtsK、MMP-9、FGF23、FGFR1、FGFR3 及 FGFR4mRNA 表达水平和蛋壳腺中钙沉积相关因子CaBP-D28k、VDR、PMCA、CA2及OPNmRNA表达水平。同时,收集各时期鸡蛋12枚常规方法检测蛋壳品质。试验结果显示:1.产蛋高峰期血清中Ca、CT、25-(OH)_2VD_3、1,25-(OH)_2VD_3和TRAP含量显著高于产蛋前期,OPN含量显著低于产蛋前期;产蛋高峰期CT、1,25-(OH)_2VD_3、E2、BGP和ALP含量显著高于产蛋后期,PICP含量明显低于产蛋后期;产蛋后期Ca、25-(OH)_2VD_3、PICP 和 TRAP含量显著高于产蛋前期,1.25-(OH)_2VD_3、E_2、OPN 和 ALP含量显著低于产蛋前期。2.产蛋前期胫骨的皮质骨结构致密,骨小梁充满整个骨髓腔,破骨细胞数量较少:产蛋高峰期骨小梁数量骤减,髓质骨增多,形成较小骨髓空腔,破骨细胞增加；产蛋后期胫骨皮质骨出现吸收腔,髓质骨减少,骨髓空腔增大,破骨细胞数量减少。3.产蛋高峰期股骨关节中FGFR3、PDGFB mRNA表达水平显著高于产蛋前期,ALP、RANKL、OPG、M-CSF、CtsK、TRAP、BMP6 和 BMP7mRNA 表达水平显著低于产蛋前期;产蛋高峰期 FGF23、V-ATPasea3、FGFR1、FGFR4 和 PDGFBmRNA 表达水平显著高于产蛋后期,M-CSF、Cts K和BMP6 mRNA表达水平显著低于产蛋后期;产蛋后期 ALP、RANKL、OPG、FGFR3、CtsK、TRAP、FGFR1 和 FGFR4mRNA 表达水平显著高产蛋前期,FGF23和V-ATPase a3 mRNA表达水平显著低于产蛋产前期。4.产蛋高峰期蛋壳腺中CaBP-D28k和VDRmRNA表达水平显著高于产蛋前期,PMCA和OPNmRNA表达水平显著低于产蛋前期;产蛋高峰期CaBP-D28k、PMCA1b、CA2和VDR mRNA表达水平显著高于产蛋后期;产蛋后期PMCA和OPN mRNA表达水平显著低于产蛋前期。5.产蛋高峰期蛋重和蛋形指数显著高于产蛋前期;产蛋高峰期蛋壳厚度、强度和蛋壳比例显著高于产蛋后期;产蛋后期蛋壳厚度、强度和蛋壳比例显著低于产蛋前期,蛋重和蛋形指数显著高于前期。本试验结果表明,整个产蛋期蛋鸡磷水平的稳定以及随着产蛋期延长的钙含量增加、骨质量和蛋壳品质降低与骨吸收增强和蛋壳腺钙沉积能力下降密切相关。
[Abstract]:In the modern intensive farming system, as a threat to health and production performance of layers of common metabolic bone disease osteoporosis in laying hens, causing serious losses to the poultry industry. A large demand for calcium during laying eggshell gland caused by the imbalance of bone formation and absorption is the main reason for the occurrence of osteoporosis in laying hens. This experiment the detection of different laying hens tibia morphological characteristics, related factors of calcium and phosphorus metabolism of bone and shell gland and the changes of eggshell quality, to explore the related factors of calcium and phosphorus metabolism of bone and shell gland changes and bone morphology and eggshell quality correlation, in order to add calcium and phosphorus of laying hens, and provide a theoretical basis for prevention of osteoporosis test. Randomly selected in the early laying period (23 weeks), during the peak period (43 weeks) and late laying period (71 weeks) of Hyline layers as the experimental object, each period of each 12, collecting blood Liquid and preparation of serum, serum Ca ratio method, P, ALP and TRAP content; radioimmunoassay of serum PTH, CT, 25- (OH) _2VD_3,1,25- (OH) _2VD_3, E_2 and BGP content; enzyme linked immunosorbent assay ELISA to detect serum PICP content. Through the acquisition of right tibia. The micro structure of micro-CT detection of bone, observe the changes of osteoclasts and bone slices were stained with TRAP. Collect femoral joint and eggshell gland, the extraction of total RNA by calcium phosphate bone detection by real-time quantitative PCR in metabolic related factors ALP, SPHK1, BMP2, BMP6, BMP7, PDGFB, TRAP, V-ATPasea3, RANKL, OPG, M-CSF. CtsK, MMP-9, FGF23, FGFR1, calcium deposition related factors CaBP-D28k, FGFR3 and the expression level of FGFR4mRNA and PMCA, VDR in the shell gland, the expression level of CA2 and OPNmRNA. At the same time, the collection period of egg 12 conventional methods for detecting eggshell quality. Experimental results showed that the serum 1. egg peak Ca, CT, 25- (OH) _2VD_3,1,25- (OH) _2VD_3 and TRAP were significantly higher than that in the early laying period, OPN content was significantly lower than that in the early laying period; laying peak CT, 1,25- (OH) _2VD_3, E2, BGP and ALP were significantly higher than the content of PICP was significantly lower than that of egg laying, egg laying; egg laying Ca, 25- (OH) _2VD_3, PICP and TRAP were significantly higher than that in the early laying period, 1.25- (OH) _2VD_3, E_2, OPN and ALP were significantly lower in cortical bone compact early laying.2. early laying of the tibia, trabecular bone to fill the bone marrow cavity, osteoclast number: egg peak number of trabecular bone marrow bone increased sharply, and the formation of small bone marrow cavity, osteoclasts increased egg laying; tibial cortical bone absorption cavity, medullary bone decreased, bone marrow cavity increases, the number of osteoclasts reduced FGFR3.3. egg peak femoral joint, the expression level of PDGFB mRNA was significantly higher than that in pre laying period, ALP, RANKL, OPG, M-CSF, CtsK, TRAP, BMP6 And the expression level of BMP7mRNA was significantly lower than that in the early laying period; laying peak FGF23, V-ATPasea3, FGFR1, expression of FGFR4 and PDGFBmRNA were significantly higher than that of egg laying, M-CSF, the expression level of Cts K and BMP6 mRNA were significantly lower than the egg laying egg laying; ALP, RANKL, OPG, FGFR3, CtsK, TRAP, FGFR1 and FGFR4mRNA expression level was significantly high egg stage the expression level of FGF23, A3 and V-ATPase mRNA were significantly lower than the level was significantly higher than that of CaBP-D28k and VDRmRNA expression in early laying egg laying peak.4. produced early eggshell glands, the expression level of PMCA and OPNmRNA were significantly lower in the early laying egg peak; CaBP-D28k, PMCA1b, CA2 and VDR mRNA expression level was significantly higher than that of egg laying; the expression level of PMCA and OPN in the late laying period mRNA.5. was significantly lower than the early laying egg peak of egg weight and egg shape index was significantly higher than that in the early laying period; the peak of egg laying shell thickness, strength The shell was significantly higher than the proportion of egg laying egg laying; eggshell thickness and eggshell strength, the proportion was significantly lower than that of early laying, egg weight and egg shape index was significantly higher than the previous. The test results show that the laying of phosphorus level stability as well as the calcium content in laying period increasing, bone quality and eggshell quality decrease is closely related with the increase of bone absorption and eggshell gland calcium deposition.

【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S831
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本文编号：1763211