發(fā)布時間：2018-04-17 10:28

  本文選題：鈣磷代謝 + 骨��； 參考：《沈陽農(nóng)業(yè)大學》2017年碩士論文

【摘要】：在現(xiàn)代集約化養(yǎng)殖體系中,蛋雞骨質(zhì)疏松癥作為一種威脅籠養(yǎng)蛋雞健康和生產(chǎn)性能的常見骨骼代謝疾病,給蛋雞養(yǎng)殖業(yè)造成了嚴重損失。產(chǎn)蛋期蛋殼腺對鈣的大量需求而引起骨形成和吸收失衡是蛋雞骨質(zhì)疏松發(fā)生的主要原因。本試驗通過檢測不同產(chǎn)蛋期蛋雞脛骨形態(tài)學特性、骨和蛋殼腺中鈣磷代謝相關(guān)因子以及蛋殼品質(zhì)的變化,探討骨和蛋殼腺中鈣磷代謝相關(guān)因子變化與骨形態(tài)及蛋殼品質(zhì)的相關(guān)性,以期為蛋雞日糧中鈣磷添加、骨質(zhì)疏松的預防提供理論依據(jù)。試驗隨機選取產(chǎn)蛋前期(23周齡)、產(chǎn)蛋高峰期(43周齡)和產(chǎn)蛋后期(71周齡)的海蘭褐蛋雞作為試驗對象,每時期各12只,采集血液并制備血清,比色法檢測血清中Ca、P、ALP和TRAP含量;放射免疫分析法檢測血清中PTH、CT、25-(OH)_2VD_3、1,25-(OH)_2VD_3、E_2和BGP含量；酶聯(lián)免疫酶免法檢測血清中PICP含量。采集右側(cè)脛骨,通過micro-CT檢測骨的微結(jié)構(gòu),骨切片TRAP染色觀察破骨細胞變化。采集股骨關(guān)節(jié)和蛋殼腺,提取總RNA,通過實時熒光定量PCR法檢測骨中鈣磷代謝相關(guān)因子ALP、SPHK1、BMP2、BMP6、BMP7、PDGFB、TRAP、V-ATPasea3、RANKL、OPG、M-CSF、CtsK、MMP-9、FGF23、FGFR1、FGFR3 及 FGFR4mRNA 表達水平和蛋殼腺中鈣沉積相關(guān)因子CaBP-D28k、VDR、PMCA、CA2及OPNmRNA表達水平。同時,收集各時期雞蛋12枚常規(guī)方法檢測蛋殼品質(zhì)。試驗結(jié)果顯示:1.產(chǎn)蛋高峰期血清中Ca、CT、25-(OH)_2VD_3、1,25-(OH)_2VD_3和TRAP含量顯著高于產(chǎn)蛋前期,OPN含量顯著低于產(chǎn)蛋前期;產(chǎn)蛋高峰期CT、1,25-(OH)_2VD_3、E2、BGP和ALP含量顯著高于產(chǎn)蛋后期,PICP含量明顯低于產(chǎn)蛋后期;產(chǎn)蛋后期Ca、25-(OH)_2VD_3、PICP 和 TRAP含量顯著高于產(chǎn)蛋前期,1.25-(OH)_2VD_3、E_2、OPN 和 ALP含量顯著低于產(chǎn)蛋前期。2.產(chǎn)蛋前期脛骨的皮質(zhì)骨結(jié)構(gòu)致密,骨小梁充滿整個骨髓腔,破骨細胞數(shù)量較少:產(chǎn)蛋高峰期骨小梁數(shù)量驟減,髓質(zhì)骨增多,形成較小骨髓空腔,破骨細胞增加；產(chǎn)蛋后期脛骨皮質(zhì)骨出現(xiàn)吸收腔,髓質(zhì)骨減少,骨髓空腔增大,破骨細胞數(shù)量減少。3.產(chǎn)蛋高峰期股骨關(guān)節(jié)中FGFR3、PDGFB mRNA表達水平顯著高于產(chǎn)蛋前期,ALP、RANKL、OPG、M-CSF、CtsK、TRAP、BMP6 和 BMP7mRNA 表達水平顯著低于產(chǎn)蛋前期;產(chǎn)蛋高峰期 FGF23、V-ATPasea3、FGFR1、FGFR4 和 PDGFBmRNA 表達水平顯著高于產(chǎn)蛋后期,M-CSF、Cts K和BMP6 mRNA表達水平顯著低于產(chǎn)蛋后期;產(chǎn)蛋后期 ALP、RANKL、OPG、FGFR3、CtsK、TRAP、FGFR1 和 FGFR4mRNA 表達水平顯著高產(chǎn)蛋前期,FGF23和V-ATPase a3 mRNA表達水平顯著低于產(chǎn)蛋產(chǎn)前期。4.產(chǎn)蛋高峰期蛋殼腺中CaBP-D28k和VDRmRNA表達水平顯著高于產(chǎn)蛋前期,PMCA和OPNmRNA表達水平顯著低于產(chǎn)蛋前期;產(chǎn)蛋高峰期CaBP-D28k、PMCA1b、CA2和VDR mRNA表達水平顯著高于產(chǎn)蛋后期;產(chǎn)蛋后期PMCA和OPN mRNA表達水平顯著低于產(chǎn)蛋前期。5.產(chǎn)蛋高峰期蛋重和蛋形指數(shù)顯著高于產(chǎn)蛋前期;產(chǎn)蛋高峰期蛋殼厚度、強度和蛋殼比例顯著高于產(chǎn)蛋后期;產(chǎn)蛋后期蛋殼厚度、強度和蛋殼比例顯著低于產(chǎn)蛋前期,蛋重和蛋形指數(shù)顯著高于前期。本試驗結(jié)果表明,整個產(chǎn)蛋期蛋雞磷水平的穩(wěn)定以及隨著產(chǎn)蛋期延長的鈣含量增加、骨質(zhì)量和蛋殼品質(zhì)降低與骨吸收增強和蛋殼腺鈣沉積能力下降密切相關(guān)。
[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.

【學位授予單位】：沈陽農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S831
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本文編號：1763211