OPG轉(zhuǎn)基因小鼠模型的建立及其骨微結(jié)構(gòu)研究
發(fā)布時(shí)間：2018-09-05 17:36
【摘要】： 目的建立OPG(osteoprotegerin,OPG)轉(zhuǎn)基因小鼠模型,觀察其骨微結(jié)構(gòu)、骨密度和骨礦含量的變化,以探討OPG在體內(nèi)調(diào)節(jié)骨代謝的作用及其分子機(jī)理。 方法構(gòu)建帶人類OPG基因啟動(dòng)子的真核表達(dá)載體pCI-hOPGp-mOPG,顯微注射入C57BL"6J小鼠受精卵原核,導(dǎo)入假孕母鼠輸卵管后培養(yǎng)子代小鼠。通過PCR法鑒定小鼠基因型,獲得OPG轉(zhuǎn)基因小鼠模型(OPG transgenic mouse ,OPG trans )。取SPF級(jí)10周齡F2代OPG trans鼠和野生型小鼠(C57BL"6J)各4只,Western Blot測定小鼠OPG蛋白表達(dá);μCT觀察小鼠脛骨近端松質(zhì)骨和皮質(zhì)骨的骨微結(jié)構(gòu);同時(shí)應(yīng)用DXA測定小鼠股骨總體骨密度和骨礦含量值。 結(jié)果1.構(gòu)建攜帶分子標(biāo)簽Flag的表達(dá)載體pCI-hOPGp-mOPG質(zhì)粒經(jīng)酶切與測序,鑒定序列正確。在69只原代小鼠中,有7只轉(zhuǎn)基因陽性Founder小鼠。目前F4代34只小鼠中,有4系共5只轉(zhuǎn)基因陽性小鼠。 2.與野生型小鼠比較,OPG轉(zhuǎn)基因小鼠脛骨松質(zhì)骨微結(jié)構(gòu)參數(shù)體積骨密度、組織骨密度、骨小梁厚度和骨小梁數(shù)目均明顯增加,而骨小梁間隔則明顯減小;皮質(zhì)骨微結(jié)構(gòu)參數(shù)皮質(zhì)骨面積、皮質(zhì)骨厚度、皮質(zhì)骨密度、皮質(zhì)骨礦含量明顯增加,而內(nèi)徑周長則明顯減小(P0.05),外徑周長間無顯著差異。股骨骨密度與骨礦含量值均明顯上升(P0.05)。 結(jié)論1.成功建立攜帶分子標(biāo)簽Flag的OPG轉(zhuǎn)基因小鼠模型。 2.OPG轉(zhuǎn)基因小鼠皮質(zhì)骨和松質(zhì)骨的骨量均增加。
[Abstract]:Objective to establish OPG (osteoprotegerin,OPG) transgenic mice model and observe the changes of bone microstructure, bone mineral density and bone mineral content in order to investigate the role of OPG in regulating bone metabolism and its molecular mechanism. Methods Eukaryotic expression vector pCI-hOPGp-mOPG, with human OPG gene promoter was constructed and microinjected into C57BL "6J mouse fertilized egg prokaryote. The mouse genotypes were identified by PCR method, and the OPG transgenic mouse model (OPG transgenic mouse OPG trans). Was obtained. The expression of OPG protein was detected by Western Blot in 4 OPG trans mice and 4 wild type mice (C57BL "6J) in F _ 2 generation of SPF grade 10 weeks old, the bone microstructure of proximal cancellous bone and cortical bone of mouse tibia was observed by 渭 CT, and the total bone mineral density and bone mineral content of femur were measured by DXA. Result 1. The expression vector pCI-hOPGp-mOPG plasmid carrying molecular label Flag was constructed by restriction endonuclease digestion and sequencing, and the sequence was confirmed to be correct. Of 69 primary mice, 7 were transgenic positive Founder mice. At present, there are 5 transgenic positive mice in 4 lines out of 34 F 4 generation mice. 2. 2. Compared with wild-type mice, the bone mineral density (BMD), tissue bone density (BMD), trabecular thickness and the number of trabeculae were significantly increased in OPG transgenic mice, while the trabecular septum decreased significantly. Cortical bone microstructure parameters: cortical bone area, cortical bone thickness, cortical bone mineral density, cortical bone mineral content increased significantly, while the inner diameter circumference decreased significantly (P0.05), but there was no significant difference between outer diameter and circumference. Bone mineral density (BMD) and bone mineral content (BMD) of femur increased significantly (P0.05). Conclusion 1. The model of OPG transgenic mice carrying molecular label Flag was successfully established. The bone mass of cortical bone and cancellous bone of 2.OPG transgenic mice were increased.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R580;R-332
本文編號(hào)：2224975
[Abstract]:Objective to establish OPG (osteoprotegerin,OPG) transgenic mice model and observe the changes of bone microstructure, bone mineral density and bone mineral content in order to investigate the role of OPG in regulating bone metabolism and its molecular mechanism. Methods Eukaryotic expression vector pCI-hOPGp-mOPG, with human OPG gene promoter was constructed and microinjected into C57BL "6J mouse fertilized egg prokaryote. The mouse genotypes were identified by PCR method, and the OPG transgenic mouse model (OPG transgenic mouse OPG trans). Was obtained. The expression of OPG protein was detected by Western Blot in 4 OPG trans mice and 4 wild type mice (C57BL "6J) in F _ 2 generation of SPF grade 10 weeks old, the bone microstructure of proximal cancellous bone and cortical bone of mouse tibia was observed by 渭 CT, and the total bone mineral density and bone mineral content of femur were measured by DXA. Result 1. The expression vector pCI-hOPGp-mOPG plasmid carrying molecular label Flag was constructed by restriction endonuclease digestion and sequencing, and the sequence was confirmed to be correct. Of 69 primary mice, 7 were transgenic positive Founder mice. At present, there are 5 transgenic positive mice in 4 lines out of 34 F 4 generation mice. 2. 2. Compared with wild-type mice, the bone mineral density (BMD), tissue bone density (BMD), trabecular thickness and the number of trabeculae were significantly increased in OPG transgenic mice, while the trabecular septum decreased significantly. Cortical bone microstructure parameters: cortical bone area, cortical bone thickness, cortical bone mineral density, cortical bone mineral content increased significantly, while the inner diameter circumference decreased significantly (P0.05), but there was no significant difference between outer diameter and circumference. Bone mineral density (BMD) and bone mineral content (BMD) of femur increased significantly (P0.05). Conclusion 1. The model of OPG transgenic mice carrying molecular label Flag was successfully established. The bone mass of cortical bone and cancellous bone of 2.OPG transgenic mice were increased.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R580;R-332
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相關(guān)期刊論文 前2條
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，本文編號(hào)：2224975
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