成纖維細(xì)胞生長因子受體3（FGFR3）對骨形成的直接調(diào)控作用
發(fā)布時(shí)間：2019-05-19 14:13
【摘要】：成纖維細(xì)胞生長因子受體(fibroblast growth factor receptors, FGFRs)屬于受體酪氨酸蛋白激酶(receptor tyrosine kinase, RTK)家族。目前已發(fā)現(xiàn)4種FGFRs,即FGFR1、FGFR2、FGFR3、FGFR4。它們之間在氨基酸水平有55%~72%的一致性。 既往研究表明,FGF/FGFRs信號通路與骨骼發(fā)育、再生和骨骼疾病有密切聯(lián)系。FGFR1、2、3功能增強(qiáng)或喪失突變可導(dǎo)致包括顱縫早閉(craniosynostosis, CS)、軟骨發(fā)育不全(achondroplasia, ACH)和CATSHL (camptodactyly, tall stature, scoliosis, and hearing loss)綜合征在內(nèi)的多種人類骨骼系統(tǒng)遺傳性疾病。 骨骼的發(fā)育是通過軟骨內(nèi)成骨(endochondral ossification)和膜內(nèi)成骨(intramembranous ossification)兩種方式來完成的。軟骨內(nèi)成骨又經(jīng)過軟骨形成(chondrogenesis)和骨形成(osteogensis)兩個(gè)密切相關(guān)的過程。而膜內(nèi)成骨則是一個(gè)單獨(dú)的骨形成的過程,即間充質(zhì)密集后直接分化為成骨細(xì)胞并成骨。 不同的FGFRs在軟骨形成和骨形成過程中的作用不一致�？偟闹v,目前認(rèn)為FGFR1、2主要調(diào)節(jié)膜內(nèi)成骨過程,而FGFR3則主要參與調(diào)控軟骨內(nèi)成骨。已發(fā)現(xiàn)人類FGFR1(P252R)功能增強(qiáng)點(diǎn)突變和多種FGFR2功能增強(qiáng)點(diǎn)突變可影響顱縫膜內(nèi)成骨過程,導(dǎo)致顱縫提前閉合,引起囟門早閉綜合征;而FGFR3功能增強(qiáng)點(diǎn)突變通過抑制生長板軟骨發(fā)育,導(dǎo)致軟骨發(fā)育不全等軟骨發(fā)育障礙性疾病。 FGFR3和FGFR1、2高度同源,也受可調(diào)節(jié)骨形成的內(nèi)源性配體FGF2、18等激活,提示FGFR3可能參與調(diào)控骨形成過程。而人類FGFR3 A391E功能增強(qiáng)型點(diǎn)突變可引起Crouzon等囟門早閉征,則是FGFR3影響骨形成的直接證據(jù)。模擬人ACH的FGFR3突變小鼠出生后15天長骨骨小梁處成骨細(xì)胞分化標(biāo)志基因Cbfa1等表達(dá)水平增高,成年期骨量減少;FGFR3敲除小鼠骨量減少,骨小梁礦化障礙。這些結(jié)果均提示FGFR3可影響成骨細(xì)胞功能和骨形成過程。 目前已有多名學(xué)者報(bào)道利用條件性基因敲除小鼠研究FGFR1、2對成骨細(xì)胞的直接調(diào)控作用,但關(guān)于FGFR3對成骨細(xì)胞及骨形成的直接調(diào)控作用及機(jī)制還不完全清楚。 骨損傷后的再生修復(fù)是局部骨骼的再發(fā)育過程。一些參與調(diào)控骨骼發(fā)育的分子如IHH/PTHrP、BMPs、Wnt等在骨再生過程中也起著重要調(diào)控作用。研究發(fā)現(xiàn),FGFR3參與調(diào)節(jié)骨損傷后的再生過程,且在一定程度上,FGFR3對骨再生和骨生長發(fā)育中的軟骨內(nèi)成骨過程可能發(fā)揮相似的負(fù)性調(diào)控作用。而Rundle等發(fā)現(xiàn)FGFR3表達(dá)于骨折部位成骨細(xì)胞和骨膜下間充質(zhì)細(xì)胞,提示FGFR3直接參與調(diào)控骨再生過程中的成骨細(xì)胞骨形成過程。那么FGFR3在骨再生過程中對成骨細(xì)胞骨形成的影響是否與FGFR3在成年期骨重建中的作用類似呢? 據(jù)此,本課題用成骨細(xì)胞特異性表達(dá)FGFR3功能增強(qiáng)點(diǎn)突變(FGFR3 K644E,Oc-cre)小鼠和模擬人軟骨發(fā)育不全的FGFR3功能增強(qiáng)點(diǎn)突變(FGFR3~(G369C/+))小鼠兩種基因敲入小鼠模型為研究對象,探討生理和病理(骨損傷)情況下,FGFR3對成骨細(xì)胞及骨形成的直接調(diào)控作用。 主要研究內(nèi)容 第一部分:成骨細(xì)胞特異性表達(dá)含功能增強(qiáng)點(diǎn)突變FGFR3對成年期小鼠骨形成的影響 1.對成骨細(xì)胞特異性表達(dá)FGFR3功能增強(qiáng)點(diǎn)突變(FGFR3K644E,Oc-cre)小鼠身長,尾長,體重等形態(tài)學(xué)指標(biāo)進(jìn)行大體觀察。 2.利用藏紅固綠染色觀察新生小鼠脛骨生長板軟骨發(fā)育情況。 3.利用X線攝片和Micro-CT觀察成年小鼠股骨骨密度及骨組織結(jié)構(gòu)的變化。 4.通過HE染色、鈣綠素雙標(biāo)實(shí)驗(yàn)觀察小鼠脛骨骨小梁的結(jié)構(gòu)及礦化情況;測定血清鈣磷含量,了解體內(nèi)鈣磷水平是否受骨骼代謝異常影響;定量PCR檢測成骨細(xì)胞分化標(biāo)志基因Cbfa1、OC、OP、Col1a1等的表達(dá)變化。 5.小鼠顱骨成骨細(xì)胞分離培養(yǎng),測定生長曲線并進(jìn)行成骨誘導(dǎo)。通過堿性磷酸酶染色、茜素紅染色觀察,結(jié)合檢測成骨相關(guān)基因Cbfa1、OC、OP、Col1a1以及FGFR1、FGFR2和BMPRIA的表達(dá),觀察成骨細(xì)胞分化和礦化情況。 6.通過TRAP染色觀察脛骨破骨細(xì)胞形成及活性。 第二部分:FGFR3功能增強(qiáng)對小鼠脛骨骨皮質(zhì)損傷修復(fù)過程的影響 1.建立FGFR3功能增強(qiáng)小鼠(FGFR3~(G369C/+))脛骨皮質(zhì)缺損模型。 2.利用HE染色和Micro-CT重建分析觀察骨再生情況。 3.定量PCR檢測新生骨組織中成骨細(xì)胞分化相關(guān)基因Cbfa1、OC、OP、Col1a1等mRNA水平表達(dá)變化。 主要實(shí)驗(yàn)結(jié)果 一. FGFR3K644E,Oc-cre小鼠骨形成增加,體型變小 1.成年期(2月齡)FGFR3K644E,Oc-cre小鼠體型變小,體重減輕;新生小鼠長骨生長板軟骨發(fā)育無異常。 2.成年期FGFR3K644E,Oc-cre小鼠骨量增多,骨形成增加X線攝片顯示FGFR3K644E,Oc-cre小鼠股骨放射密度增高;Micro-CT掃描顯示FGFR3K644E,Oc-cre小鼠BV/TV,骨小梁數(shù)量( Tb.N)及骨小梁厚度( Tb.Th)均顯著升高;HE染色FGFR3K644E,Oc-cre小鼠干骺端和骨骺部位骨小梁較野生小鼠明顯增多、增長、增粗。提示成年期FGFR3K644E,Oc-cre骨量增多,骨形成增加。 3. FGFR3K644E,Oc-cre小鼠成骨細(xì)胞分化增強(qiáng),礦化障礙 1) 2月齡FGFR3K644E,Oc-cre小鼠脛骨骨組織中成骨細(xì)胞分化標(biāo)志基因Cbfa1、OP和Col1a1表達(dá)上調(diào),表明成骨細(xì)胞分化增強(qiáng);鈣綠素雙標(biāo)實(shí)驗(yàn)提示FGFR3 K644E,Oc-cre小鼠礦化能力減弱;體內(nèi)血清鈣磷含量正常,提示骨骼礦化障礙沒有影響全身鈣磷水平。 2) FGFR3K644E,Oc-cre小鼠顱骨成骨細(xì)胞增殖減慢;經(jīng)成骨誘導(dǎo)培養(yǎng)后堿性磷酸酶表達(dá)增加,鈣結(jié)節(jié)形成減少;成骨細(xì)胞分化標(biāo)志基因Cbfa1、OP、OC、Col1a1表達(dá)均顯著上調(diào),表明成骨細(xì)胞分化增強(qiáng),礦化降低。 3) FGFR3K644E,Oc-cre小鼠成骨相關(guān)基因FGFR1、FGFR2及BMPRIA表達(dá)下調(diào)。 4.FGFR3K644E,Oc-cre小鼠破骨細(xì)胞骨吸收作用沒有改變TRAP染色后計(jì)數(shù)結(jié)果顯示FGFR3K644E,Oc-cre小鼠脛骨單位骨小梁面積上的破骨細(xì)胞數(shù)量與野生小鼠無明顯差別,提示FGFR3K644E,Oc-cre小鼠破骨細(xì)胞數(shù)量及功能無明顯改變。 二. FGFR3~(G369C/+)小鼠脛骨骨皮質(zhì)損傷愈合延遲 1. FGFR3~(G369C/+)小鼠骨皮質(zhì)損傷后局部新生骨組織增多Micro-CT掃描重建分析及HE染色顯示FGFR3~(G369C/+)小鼠損傷局部新生骨組織數(shù)量和體積較野生小鼠增加;新生骨組織成骨細(xì)胞分化標(biāo)記基因Cbfa1、OP等表達(dá)顯著上調(diào),提示成骨細(xì)胞分化增強(qiáng)。 2. FGFR3~(G369C/+)小鼠皮質(zhì)骨愈合延遲MicroCT掃描重建分析及HE染色顯示在損傷后第21天,野生型小鼠和FGFR3~(G369C/+)小鼠在皮質(zhì)缺損處都出現(xiàn)了新生板層骨,但FGFR3~(G369C/+)小鼠的新生骨皮質(zhì)厚度要小于野生小鼠。 主要結(jié)論 1.成骨細(xì)胞特異性表達(dá)FGFR3功能增強(qiáng)點(diǎn)突變小鼠體型變小,但生長板軟骨未見明顯異常,提示成骨異常在ACH侏儒體型發(fā)生中有一定作用。 2.成骨細(xì)胞特異性表達(dá)FGFR3功能增強(qiáng)點(diǎn)突變小鼠成年期骨形成增加,導(dǎo)致骨量增多。 3. FGFR3功能增強(qiáng)促進(jìn)成骨細(xì)胞分化,抑制其礦化。 4. FGFR3功能增強(qiáng)對成骨細(xì)胞的影響可能部分是通過FGFR1、FGFR2及BMPRIA的繼發(fā)性改變所致。 5. FGFR3功能增強(qiáng)導(dǎo)致小鼠骨皮質(zhì)缺損愈合延遲。
[Abstract]:Fibroblast growth factor receptors (FGFRs) belong to the receptor tyrosine kinase (RTK) family. Four FGFRs, FGFR1, FGFR2, FGFR3, and FGFR4 have been found. And the amino acid level is between 55 and 72 percent. Previous studies have shown that the FGF/ FGFRs signaling pathway is closely related to bone development, regeneration and skeletal diseases Contact. FGFR1,2,3 functional enhancement or loss of mutations can lead to a variety of human skeletal system heritages, including cranio-early-closure (CS), achondroplasia, ACH, and CATSHL syndrome The development of the bone is through the endochondral bone and the intramembranous ossification. The endochondral bone, in turn, is formed by the formation of the cartilage and the bone formation (ostegosis). The process of bone formation is the process of the formation of a single bone, that is, the direct differentiation of the mesenchyme into the bone. The cells are not osteogenic. Different FGFRs are formed in the form of cartilage and bone. In general, FGFR1,2 mainly regulate the osteogenesis in the membrane, while FGFR3 is the main parameter. It has been found that human FGFR1 (P252R) function-enhanced point mutation and a variety of FGFR2 function-enhanced point mutations can affect the osteogenic process in the cranio-suture membrane, resulting in premature closure of the cranial suture, resulting in an early-closure syndrome, while the FGFR3 function-enhancing point mutation is through inhibition The development of the cartilage of the growth plate, which leads to the soft development of the cartilage, etc. Bone developmental disorders. FGFR3 and FGFR1,2 are highly homologous, and are also activated by the endogenous ligands FGF2,18, which can modulate bone formation, suggesting that FGFR3 It is possible to participate in the regulation of the bone formation process, while the human FGFR3 A391E functional enhancement point mutation can cause early closure of the Crouzon et al., and is the FGFR 3. Direct evidence of the formation of bone. The expression level of the osteoblast differentiation marker gene Cbfa1 was increased at 15 days after the birth of the FGFR3 mutant of the human ACH, and the expression level of the osteoblast differentiation marker gene Cbfa1 in the bone trabecula at 15 days after the birth of the mouse was increased, and the amount of the adult bone decreased; and the FGFR3 knocked out the mouse bone. The amount of bone trabecula is reduced, and the bone trabeculectomy is an obstacle. These results suggest that FGFR3 can be affected. Osteoclasts function and bone formation process. Several scholars have reported that using the conditional gene knockout mice to study the direct regulatory action of FGFR1,2 on osteoblasts, but with respect to the direct effect of FGFR3 on the formation of osteoblasts and bone The action and mechanism of the grafting are not completely clear. The bone injury The post-regeneration repair is the re-development of the local bone. Some molecules involved in the control of bone development, such as IHH/ PTHrP, BMPs, Wnt, It is found that FGFR3 is involved in the regeneration of bone regeneration and, to some extent, FGFR3 plays an important role in the regeneration of bone and in the growth and development of bone. The osteogenic process may play a similar negative control role, and Rundle et al. found that FGFR3 is expressed in the bone and subperiosteal mesenchymal cells of the fracture site, suggesting that FGFR3 is directly involved in the regulation. In the process of bone regeneration, the effect of FGFR3 on the bone formation of osteoblasts in the process of bone regeneration is similar to that of FG The role of FR3 in the reconstruction of the adult bone is similar, according to which the function of FGFR3 functional enhancement point mutation (FGFR3 K644E, Oc-cre) and the FGFR3 functional enhancement point mutation (FGFR3-(G369C/ +) The two genes of the mouse were knocked into the mouse model to study the object, and the physiological and pathological changes (bone injury) were discussed. GFR3 pair to Direct regulatory effects of osteocytes and bone formation. Part 1: Osteoblastic specificity Effect of the expression of FGFR3 with functional enhancement point mutation on bone formation in adult mice 1. The expression of FGFR3 in the osteoblast-specific expression was enhanced. Point mutation (FGFR3 K644E, The morphology of Oc-cre (Oc-cre) mouse's body length, tail length, body weight, etc. A.2. The development of the growth plate of the tibial growth plate of the newly-born mouse was observed by using the red solid green staining. Changes of bone mineral density and bone structure in adult mice were observed by X-ray film and Micro-CT.4. The structure and mineralization of the bone trabecula of the tibia were observed by HE staining and calcium-green-green double-scale experiment. The content of serum calcium and phosphorus was determined. Affected by bone metabolism; quantitative P CR detection of osteoblast differentiation marker gene Cbfa1, OC, OP, Chol 1. The expression changes of 1a1 and so on.5. The mouse's skull osteoblast was isolated and cultured, the growth curve was determined and the osteogenic induction was performed. The bone formation-related genes Cbfa1, OC, OP, Co were detected by the staining of alkaline phosphatase and the red staining of the bone. l1a1 and FGFR1, FGFR2, and BMPRIA The expression of TRAP was used to study the differentiation and mineralization of the osteoblast. To observe the formation and activity of osteoclast in the tibia. The second part: the function of FGFR3 enhances the damage to the cortical bone of the tibia. Effect of the repair process 1. Establishment of a FGFR3 function-enhancing mouse (FGF 3 ~ (G369C/ +)) tibial cortical defect model.2. The bone regeneration was observed by HE staining and micro-CT reconstruction. 3. Quantitation PC R. Detection of the Osteoblastic Differentiation-related Genes Cbfa1, OC, O in the New Bone Tissue Changes of mRNA level in P, Col1a1, etc. The main results are as follows: 1.FGFR3 K644E, Oc-cre mouse bone formation The body size decreased 1. The size of the FGFR3 K644E and Oc-cre mice became smaller and weight loss in the adult mice.2. The growth of the growth plate of the long bone in the new mice was not abnormal.2. The bone mass of FGFR3 K644E and Oc-cre mice increased, and the bone formation increased the X-ray film to show the increase of the radial density of the femur in the Oc-cre mice, and the micro-CT scan showed F. GFR3K644E, Oc-cre mice BV/ TV, trabecular number (Tb. N) and the thickness of the bone trabecula (Tb. Th) increased significantly; the HE staining of FGFR3 K644E In the Oc-cre mice, the bone trabecula of the dry and bone parts of the mouse was significantly increased, the growth and the increase of the bone trabecula were increased. 2-month-old FGFR3 K644E, Oc-cre mouse osteoblast differentiation enhancement, mineralization disorder 1)2-month-old FGFR3 K644E, Oc-cre mouse tibia bone tissue Up-regulation of expression of Cbfa1, OP and Col1a1 in bone cell differentiation marker genes, and Table The results showed that the bone mineralizing ability of FGFR3 K644E and Oc-cre mice was weakened, and the serum calcium and phosphorus content in the body was normal, suggesting that the bone mineralization disorder did not affect the whole body calcium and phosphorus level. The expression of alkaline phosphatase after osteoinductive culture is increased, and the formation of calcium nodules is reduced; and the osteoblast differentiation marker gene Cbfa1, OP, OC, Co The expression of l1a1 was up-regulated, indicating that the osteoblast differentiation was enhanced and the mineralization decreased.3) The expression of FGFR3 K644E, Oc-cre osteogenic related genes FGFR1, FGFR2 and BMPRIA was down-regulated. show F GFR3K644E, Oc-cre mouse tibial unit bone trabecula area No significant difference was found between the number of osteoclasts and wild mice, suggesting that the number and function of osteoclast in FGFR3 K644E and Oc-cre mice did not change significantly. Micro-CT scan and reconstruction of locally-born bone tissue after bone-cortical injury in FR3-(G369C/ +) mice Analysis and HE staining showed that the number and volume of local new bone tissue in FGFR3-(G369C/ +) mice increased significantly, and the expression of Cbfa1, OP and so on in new bone tissue was up-regulated, suggesting that the differentiation of osteoblasts was enhanced. reconstruction and analysis and HE staining showed a new lamellar bone in both wild-type and FGFR3-(G369C/ +) mice at the 21-day post-injury, but FGFR3-(G36 9 C/ +) The thickness of the new bone cortex of the mice is less than that of the wild mice. The expression of FGFR3 functional enhancement point mutant mice was small, but the growth plate There was no obvious abnormality in the cartilage, suggesting that the osteogenic abnormality could play a role in the body type of the ACH dwarf.2. The osteoblast-specific expression of the FGFR3 function The increase of the bone formation in the formation of the enhanced point mutant mice leads to an increase in bone mass.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363
[Abstract]:Fibroblast growth factor receptors (FGFRs) belong to the receptor tyrosine kinase (RTK) family. Four FGFRs, FGFR1, FGFR2, FGFR3, and FGFR4 have been found. And the amino acid level is between 55 and 72 percent. Previous studies have shown that the FGF/ FGFRs signaling pathway is closely related to bone development, regeneration and skeletal diseases Contact. FGFR1,2,3 functional enhancement or loss of mutations can lead to a variety of human skeletal system heritages, including cranio-early-closure (CS), achondroplasia, ACH, and CATSHL syndrome The development of the bone is through the endochondral bone and the intramembranous ossification. The endochondral bone, in turn, is formed by the formation of the cartilage and the bone formation (ostegosis). The process of bone formation is the process of the formation of a single bone, that is, the direct differentiation of the mesenchyme into the bone. The cells are not osteogenic. Different FGFRs are formed in the form of cartilage and bone. In general, FGFR1,2 mainly regulate the osteogenesis in the membrane, while FGFR3 is the main parameter. It has been found that human FGFR1 (P252R) function-enhanced point mutation and a variety of FGFR2 function-enhanced point mutations can affect the osteogenic process in the cranio-suture membrane, resulting in premature closure of the cranial suture, resulting in an early-closure syndrome, while the FGFR3 function-enhancing point mutation is through inhibition The development of the cartilage of the growth plate, which leads to the soft development of the cartilage, etc. Bone developmental disorders. FGFR3 and FGFR1,2 are highly homologous, and are also activated by the endogenous ligands FGF2,18, which can modulate bone formation, suggesting that FGFR3 It is possible to participate in the regulation of the bone formation process, while the human FGFR3 A391E functional enhancement point mutation can cause early closure of the Crouzon et al., and is the FGFR 3. Direct evidence of the formation of bone. The expression level of the osteoblast differentiation marker gene Cbfa1 was increased at 15 days after the birth of the FGFR3 mutant of the human ACH, and the expression level of the osteoblast differentiation marker gene Cbfa1 in the bone trabecula at 15 days after the birth of the mouse was increased, and the amount of the adult bone decreased; and the FGFR3 knocked out the mouse bone. The amount of bone trabecula is reduced, and the bone trabeculectomy is an obstacle. These results suggest that FGFR3 can be affected. Osteoclasts function and bone formation process. Several scholars have reported that using the conditional gene knockout mice to study the direct regulatory action of FGFR1,2 on osteoblasts, but with respect to the direct effect of FGFR3 on the formation of osteoblasts and bone The action and mechanism of the grafting are not completely clear. The bone injury The post-regeneration repair is the re-development of the local bone. Some molecules involved in the control of bone development, such as IHH/ PTHrP, BMPs, Wnt, It is found that FGFR3 is involved in the regeneration of bone regeneration and, to some extent, FGFR3 plays an important role in the regeneration of bone and in the growth and development of bone. The osteogenic process may play a similar negative control role, and Rundle et al. found that FGFR3 is expressed in the bone and subperiosteal mesenchymal cells of the fracture site, suggesting that FGFR3 is directly involved in the regulation. In the process of bone regeneration, the effect of FGFR3 on the bone formation of osteoblasts in the process of bone regeneration is similar to that of FG The role of FR3 in the reconstruction of the adult bone is similar, according to which the function of FGFR3 functional enhancement point mutation (FGFR3 K644E, Oc-cre) and the FGFR3 functional enhancement point mutation (FGFR3-(G369C/ +) The two genes of the mouse were knocked into the mouse model to study the object, and the physiological and pathological changes (bone injury) were discussed. GFR3 pair to Direct regulatory effects of osteocytes and bone formation. Part 1: Osteoblastic specificity Effect of the expression of FGFR3 with functional enhancement point mutation on bone formation in adult mice 1. The expression of FGFR3 in the osteoblast-specific expression was enhanced. Point mutation (FGFR3 K644E, The morphology of Oc-cre (Oc-cre) mouse's body length, tail length, body weight, etc. A.2. The development of the growth plate of the tibial growth plate of the newly-born mouse was observed by using the red solid green staining. Changes of bone mineral density and bone structure in adult mice were observed by X-ray film and Micro-CT.4. The structure and mineralization of the bone trabecula of the tibia were observed by HE staining and calcium-green-green double-scale experiment. The content of serum calcium and phosphorus was determined. Affected by bone metabolism; quantitative P CR detection of osteoblast differentiation marker gene Cbfa1, OC, OP, Chol 1. The expression changes of 1a1 and so on.5. The mouse's skull osteoblast was isolated and cultured, the growth curve was determined and the osteogenic induction was performed. The bone formation-related genes Cbfa1, OC, OP, Co were detected by the staining of alkaline phosphatase and the red staining of the bone. l1a1 and FGFR1, FGFR2, and BMPRIA The expression of TRAP was used to study the differentiation and mineralization of the osteoblast. To observe the formation and activity of osteoclast in the tibia. The second part: the function of FGFR3 enhances the damage to the cortical bone of the tibia. Effect of the repair process 1. Establishment of a FGFR3 function-enhancing mouse (FGF 3 ~ (G369C/ +)) tibial cortical defect model.2. The bone regeneration was observed by HE staining and micro-CT reconstruction. 3. Quantitation PC R. Detection of the Osteoblastic Differentiation-related Genes Cbfa1, OC, O in the New Bone Tissue Changes of mRNA level in P, Col1a1, etc. The main results are as follows: 1.FGFR3 K644E, Oc-cre mouse bone formation The body size decreased 1. The size of the FGFR3 K644E and Oc-cre mice became smaller and weight loss in the adult mice.2. The growth of the growth plate of the long bone in the new mice was not abnormal.2. The bone mass of FGFR3 K644E and Oc-cre mice increased, and the bone formation increased the X-ray film to show the increase of the radial density of the femur in the Oc-cre mice, and the micro-CT scan showed F. GFR3K644E, Oc-cre mice BV/ TV, trabecular number (Tb. N) and the thickness of the bone trabecula (Tb. Th) increased significantly; the HE staining of FGFR3 K644E In the Oc-cre mice, the bone trabecula of the dry and bone parts of the mouse was significantly increased, the growth and the increase of the bone trabecula were increased. 2-month-old FGFR3 K644E, Oc-cre mouse osteoblast differentiation enhancement, mineralization disorder 1)2-month-old FGFR3 K644E, Oc-cre mouse tibia bone tissue Up-regulation of expression of Cbfa1, OP and Col1a1 in bone cell differentiation marker genes, and Table The results showed that the bone mineralizing ability of FGFR3 K644E and Oc-cre mice was weakened, and the serum calcium and phosphorus content in the body was normal, suggesting that the bone mineralization disorder did not affect the whole body calcium and phosphorus level. The expression of alkaline phosphatase after osteoinductive culture is increased, and the formation of calcium nodules is reduced; and the osteoblast differentiation marker gene Cbfa1, OP, OC, Co The expression of l1a1 was up-regulated, indicating that the osteoblast differentiation was enhanced and the mineralization decreased.3) The expression of FGFR3 K644E, Oc-cre osteogenic related genes FGFR1, FGFR2 and BMPRIA was down-regulated. show F GFR3K644E, Oc-cre mouse tibial unit bone trabecula area No significant difference was found between the number of osteoclasts and wild mice, suggesting that the number and function of osteoclast in FGFR3 K644E and Oc-cre mice did not change significantly. Micro-CT scan and reconstruction of locally-born bone tissue after bone-cortical injury in FR3-(G369C/ +) mice Analysis and HE staining showed that the number and volume of local new bone tissue in FGFR3-(G369C/ +) mice increased significantly, and the expression of Cbfa1, OP and so on in new bone tissue was up-regulated, suggesting that the differentiation of osteoblasts was enhanced. reconstruction and analysis and HE staining showed a new lamellar bone in both wild-type and FGFR3-(G369C/ +) mice at the 21-day post-injury, but FGFR3-(G36 9 C/ +) The thickness of the new bone cortex of the mice is less than that of the wild mice. The expression of FGFR3 functional enhancement point mutant mice was small, but the growth plate There was no obvious abnormality in the cartilage, suggesting that the osteogenic abnormality could play a role in the body type of the ACH dwarf.2. The osteoblast-specific expression of the FGFR3 function The increase of the bone formation in the formation of the enhanced point mutant mice leads to an increase in bone mass.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363
【共引文獻(xiàn)】
相關(guān)期刊論文 前8條
1 周銳;蘇楠;李燦;陳思宇;謝楊麗;陳林;;小鼠脛骨骨皮質(zhì)損傷修復(fù)過程中成纖維生長因子受體的表達(dá)[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2011年10期
2 吳莉萍;凌均h，
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