基于破骨細(xì)胞功能探討腺苷激酶敲除小鼠和貞術(shù)調(diào)脂膠囊對(duì)骨代謝的影響
發(fā)布時(shí)間：2018-09-13 21:35
【摘要】：【目的】我們?cè)跇?gòu)建髓性單核細(xì)胞敲除腺苷激酶(Adenosine Kinase,ADK)的小鼠過(guò)程中,與對(duì)照組小鼠相比,顯示體重明顯下降。因此,我們將利用髓性單核細(xì)胞系特異性敲除ADK小鼠進(jìn)行破骨細(xì)胞(osteoclast,OC)分化和骨吸收功能的研究,探討髓性單核細(xì)胞敲除ADK對(duì)骨代謝的影響。同時(shí)在建立的破骨細(xì)胞體外培養(yǎng)體系下,評(píng)價(jià)貞術(shù)調(diào)脂膠囊(FTZ)對(duì)破骨細(xì)胞分化和骨吸收功能的影響�！痉椒ā�1.髓性單核細(xì)胞敲除ADK小鼠的鑒定方法選取雜交繁殖后的子代鼠尾進(jìn)行PCR基因型分析鑒定,篩選出ADKf/f、ADKf/flysmcre/cre、ADKf/flysmcre/+三種基因敲除小鼠。采用Western-blot技術(shù)檢測(cè)不同基因型小鼠原代巨噬細(xì)胞ADK蛋白的表達(dá)量。2.體外破骨細(xì)胞分化及骨吸收功能檢測(cè)方法利用ADKf/f、ADKf/flysmcre/cre、ADKf/flysmcre/+三種基因敲除小鼠,體外獲取分離小鼠的原代骨髓單核細(xì)胞,在M-CSF和RANKL的刺激下誘導(dǎo)培養(yǎng)破骨細(xì)胞,同時(shí)收集野生型BL/6J小鼠原代骨髓單核細(xì)胞并給予ADK抑制劑(ABT-702)干預(yù),進(jìn)行抗酒石酸酸性磷酸酶特異性Trap染色、骨吸收實(shí)驗(yàn)、骨代謝功能相關(guān)TRAP、NFATc1、c-Fos、MMP-9、Cts K、c-Src、CTR基因RT-PCR檢測(cè),評(píng)價(jià)髓性單核細(xì)胞敲除ADK對(duì)體外破骨細(xì)胞分化及骨吸收功能的作用。3.基因型小鼠骨形態(tài)計(jì)量學(xué)檢測(cè)方法比較4月齡ADKf/f、ADKf/flysmcre/cre、ADKf/flysmcre/+雄性小鼠的體重、股骨濕重;收集脛骨上、中段組織,采用HE染色觀察小鼠骨組織病理形態(tài)學(xué)改變,通過(guò)骨組織靜態(tài)學(xué)分析系統(tǒng)分析小鼠體內(nèi)骨量的變化。4.FTZ對(duì)體外破骨細(xì)胞分化和骨吸收的影響利用已建立的破骨細(xì)胞體外培養(yǎng)體系,收集野生型BL/6J小鼠原代骨髓單核細(xì)胞,并添加大鼠含藥血清干預(yù),進(jìn)行抗酒石酸酸性磷酸酶特異性TRAP染色、骨吸收實(shí)驗(yàn)、骨代謝功能相關(guān)TRAP、NFATc1、c-Fos、MMP-9、Cts K、c-Src、CTR基因RT-PCR檢測(cè),評(píng)價(jià)FTZ對(duì)破骨細(xì)胞分化和吸收功能的作用�！窘Y(jié)果】1.ADKf/flysmcre/cre小鼠ADK的敲除效率比ADKf/fLysmcre/+小鼠、ADKf/f小鼠高。(1)PCR檢測(cè)鼠尾基因型鑒定結(jié)果:ADK純合子條帶為360bp;ADKf/f小鼠在350bp處有1條清晰條帶,ADKf/flysmcre/cre小鼠在700bp處有一條清晰條帶,ADKf/flysmcre/+小鼠在350bp、700bp處有兩條條帶。(2)Western blot檢測(cè)髓性單核細(xì)胞ADK蛋白水平結(jié)果:與ADKf/f組相比較,ADKf/fLysmcre/cre組ADK蛋白表達(dá)水平降低69.77%(P0.01),ADKf/fLysmcre/+組ADK蛋白表達(dá)水平降低36.83%(P0.05)。2.髓性單核細(xì)胞敲除ADK小鼠體外破骨細(xì)胞分化增多,骨吸收功能增強(qiáng)。(1)髓性單核細(xì)胞敲除ADK小鼠增加了破骨細(xì)胞分化:不同基因型小鼠體外破骨細(xì)胞培養(yǎng)結(jié)果發(fā)現(xiàn),與ADKf/f組相比,ADKf/fLysmcre/cre組的破骨細(xì)胞數(shù)量較多,具有顯著性差異(P0.01);但ADKf/f Lysmcre/+組的破骨細(xì)胞數(shù)量無(wú)明顯變化(P0.05);與ADKf/fLysmcre/cre組相比,ADKf/fLysmcre/+組的破骨細(xì)胞數(shù)量有顯著性差異(P0.01)。ABT-702干預(yù)破骨細(xì)胞培養(yǎng)結(jié)果發(fā)現(xiàn),分別與control組、5%Et OH組相比,100n M組的破骨細(xì)胞數(shù)量較多,有顯著性差異(P0.01)。破骨細(xì)胞分化相關(guān)基因RT-PCR檢測(cè)結(jié)果,相比于ADKf/f組,ADKf/fLysmcre/cre組的TRAP(P0.01)、NFATc1(P0.05)基因表達(dá)量增加;c-Fos基因表達(dá)量無(wú)差異(P0.05),但ADKf/f Lysmcre/+組上述基因無(wú)明顯變化(P0.05);與ADKf/f Lysmcre/cre組相比,ADKf/fLysmcre/+組的TRAP和NFATc1基因表達(dá)量有統(tǒng)計(jì)學(xué)差異(P0.05),c-Fos基因表達(dá)量無(wú)差異(P0.05)。ABT-702干預(yù)破骨細(xì)胞分化相關(guān)基因RT-PCR結(jié)果顯示,相比于5%Et OH組,100n M組的TRAP、NFATc1的m RNA基因表達(dá)量增多,具有統(tǒng)計(jì)學(xué)差異(P0.05);c-Fos基因表達(dá)量無(wú)顯著性差異(P0.05)。(2)髓性單核細(xì)胞敲除ADK增強(qiáng)了破骨細(xì)胞吸收功能:不同基因型小鼠體外破骨細(xì)胞骨吸收實(shí)驗(yàn)結(jié)果發(fā)現(xiàn),與ADKf/f組相比,ADKf/fLysmcre/cre組培養(yǎng)的破骨細(xì)胞的骨陷窩面積顯著增大(P0.01)和數(shù)量增多(P0.05),ADKf/f Lysmcre/+組的破骨細(xì)胞骨陷窩面積和數(shù)量比較,無(wú)明顯變化(P0.05);與ADKf/f Lysmcre/cre組相比,ADKf/f Lysmcre/+組的破骨細(xì)胞骨陷窩面積和數(shù)量均具有顯著性差異(P0.01)。ABT-702干預(yù)破骨細(xì)胞骨吸收實(shí)驗(yàn)結(jié)果發(fā)現(xiàn),與5%Et OH組相比,100n M骨陷窩面積增大和數(shù)量增多,有顯著性差異(P0.01)。破骨細(xì)胞吸收功能相關(guān)基因RT-PCR檢測(cè)結(jié)果,相比于ADKf/f組,ADKf/fLysmcre/cre組的MMP-9、Cts K、CTR基因表達(dá)量增多,具有顯著性差異(P0.01),c-Src基因表達(dá)量無(wú)顯著性差異(P0.05),ADKf/f Lysmcre/+組的Cts K基因表達(dá)量有統(tǒng)計(jì)學(xué)差異(P0.05);與ADKf/f Lysmcre/cre組相比,ADKf/f Lysmcre/+組的MMP-9、Cts K、CTR基因表達(dá)量具有統(tǒng)計(jì)學(xué)差異(P0.05)。ABT-702干預(yù)破骨細(xì)胞骨吸收相關(guān)基因結(jié)果顯示,相比于5%Et OH組,100n M組的MMP-9(P0.01)、Cts K(P0.05)基因表達(dá)量增多,但c-Src、CTR基因表達(dá)量無(wú)明顯變化(P0.05)。3.髓性單核細(xì)胞敲除ADK小鼠體內(nèi)骨量的變化相比于ADKf/f小鼠,ADKf/flysmcre/cre小鼠體重和股骨濕重最輕,具有顯著性差異(P0.01),ADKf/flysmcre/+小鼠無(wú)明顯變化(P0.05);與ADKf/f Lysmcre/cre小鼠相比,ADKf/fLysmcre/+小鼠體重和股骨重量均有顯著性差異(P0.01)。ADKf/f組松質(zhì)骨部分骨小梁豐富呈淺紫粉紅色,結(jié)構(gòu)致密粗實(shí),連續(xù)性好呈網(wǎng)狀,ADKf/fLysmcre/cre組骨小梁明顯減少,排列稀疏,斷裂狀,ADKf/fLysmcre/+組骨小梁結(jié)構(gòu)介于兩組間,骨小梁細(xì)長(zhǎng),部分?jǐn)嗔�。骨組織靜態(tài)參數(shù)結(jié)果顯示,ADKf/flysmcre/cre小鼠分別與ADKf/f小鼠、ADKf/flysmcre/+小鼠相比,%Tb.Ar、Tb.Th、Tb.N、%Ob.S/BS、Ob.N/BS、Tb.Sp、%Oc.S/BS、Oc.N/BS均有統(tǒng)計(jì)學(xué)差異(P0.01;P0.05)。同樣,脛骨中段分析,ADKf/f Lysmcre/cre組與其余兩組計(jì)較,%Ct.Ar、%Ma.Ar參數(shù)變化明顯(P0.01)。4.FTZ抑制了破骨細(xì)胞分化和吸收功能不同濃度劑量對(duì)破骨細(xì)胞有不一樣的抑制作用,與control組相比,Alendronate組、FTZ-H組和FTZ-M組的破骨細(xì)胞數(shù)量(P0.01)、骨陷窩面積(P0.05)和骨陷窩數(shù)量均減少(P0.01),比FTZ-L組的破骨細(xì)胞數(shù)量少(P0.05)和骨陷窩數(shù)量少(FTZ-H組P0.01;FTZ-M組P0.05);與Alendronate組相比,FTZ-L組的骨陷窩數(shù)量具有顯著性差異(P0.01)。破骨細(xì)胞分化相關(guān)基因RT-PCR結(jié)果顯示,相比于control組,FTZ-H組的TRAP、NFATc1基因表達(dá)量明顯降低(P0.05),c-Fos基因表達(dá)量無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。骨吸收相關(guān)基因表達(dá)檢測(cè)結(jié)果顯示,與control組相比,FTZ-H組MMP-9基因表達(dá)量下降,具有統(tǒng)計(jì)學(xué)差異(P0.01),Cts K、c-Src、CTR基因表達(dá)量無(wú)明顯變化(P0.05)�！窘Y(jié)論】1、髓性單核細(xì)胞敲除ADK小鼠增強(qiáng)了破骨細(xì)胞分化功能。2、髓性單核細(xì)胞敲除ADK小鼠增強(qiáng)了破骨細(xì)胞吸收功能。3、髓性單核細(xì)胞細(xì)胞敲除ADK小鼠顯示骨量下降,骨結(jié)構(gòu)異常。4、FTZ改善骨質(zhì)疏松通過(guò)抑制破骨細(xì)胞的分化和吸收功能。
[Abstract]:[Objective] Compared with the control group, the weight of ADK mice was significantly decreased during the construction of ADK knockout mice. Therefore, the osteoclast (OC) differentiation and bone resorption function of ADK mice were studied by using myelomonocyte line specific knockout mice. The effect of sexual monocyte knockout ADK on bone metabolism was evaluated in vitro. The effects of Zhenzhu Tiaozhi capsule (FTZ) on osteoclast differentiation and bone resorption were evaluated under the established osteoclast culture system. ADK f/f, ADK f/flysmcre/cre, ADK f/flysmcre/+ three gene knockout mice were identified and screened. The expression of ADK protein in primary macrophages of different genotypes of mice was detected by Western blot. 2. Osteoclast differentiation and bone resorption function in vitro were detected by ADK f/f, ADK f/flysmcre/cre, ADK f/flysmcre/+ three gene knockout mice. Osteoclasts were induced by M-CSF and RANKL stimulation. Primary bone marrow mononuclear cells of wild type BL/6J mice were collected and interfered with ADK inhibitor (ABT-702), tartrate-resistant acid phosphatase specific Trap staining, bone resorption test, TRAP, NFATc1 related to bone metabolism were performed. The effects of myelomonocyte knockout ADK on osteoclast differentiation and bone resorption in vitro were evaluated by RT-PCR. 3. Bone morphometry of 4-month-old ADK f/f, ADK f/flysmcre/cre, ADK f/flysmcre/+ male mice was compared with that of 4-month-old ADK f/f, ADK f/flysmcre/+ male mice. HE staining was used to observe the pathomorphological changes of bone tissue in mice. The changes of bone mass in mice were analyzed by bone histostatic analysis system. 4. The effect of FTZ on osteoclast differentiation and bone resorption in vitro. The primary bone marrow mononuclear cells of wild type BL/6J mice were collected and added into the culture system of osteoclasts in vitro. Drug serum intervention, tartrate-resistant acid phosphatase-specific TRAP staining, bone resorption test, Bone Metabolism-Related TRAP, NFATc1, c-Fos, MMP-9, Cts K, c-Src, CTR gene RT-PCR assay were used to evaluate the effect of FTZ on osteoclast differentiation and absorption function. [Results] 1. The knockout efficiency of ADKf/smcre/cre/ADflyK was higher than that of ADKf/fLysms/+mice. ADK f / flysmcre / CRE mice had a clear band at 350 bp. ADK f / flysmcre / CRE mice had a clear band at 700 bp. ADK f / flysmcre / + mice had two bands at 350 BP and 700 bp. (2) Western blot was used to detect ADK protein level in myeloid monocytes. Compared with ADK f/f group, ADK protein expression in ADK f/fLysmcre/cre group was decreased by 69.77% (P 0.01), and ADK protein expression in ADK f/fLysmcre/+ group was decreased by 36.83% (P 0.05). 2. Osteoclast differentiation and bone resorption were increased in ADK-knockout mice. (1) Osteoclast differentiation was increased in ADK-knockout mice: no The results of osteoclasts culture in vitro showed that the number of osteoclasts in ADKf/fLysmcre/cre group was significantly higher than that in ADKf/fLysmcre/cre group (P 0.01), but there was no significant change in the number of osteoclasts in ADKf/fLysmcre/+ group (P 0.05). The results showed that the number of osteoclasts in 100nM group was higher than that in control group and 5% Et OH group (P 0.01). The expression of NFATc1 (P 0.05) gene in ADKf / fLysmcre / CRE group was higher than that in ADKf / F group by RT-PCR. There was no significant difference in the expression of OS gene (P 0.05), but there was no significant change in the above genes in ADKf / fLysmcre /+ group (P 0.05). Compared with ADKf / fLysmcre / cre group, the expression of TRAP and NFATc1 gene in ADKf / fLysmcre /+ group was significantly different (P 0.05), and there was no difference in the expression of c-Fos gene (P 0.05). The results of RT-PCR showed that ABT-702 intervened osteoclast differentiation-related genes. Compared with 5% Et OH group, the expression of TRAP and NFATc1 m RNA in 100N M group increased, with statistical difference (P 0.05); the expression of c-Fos gene had no significant difference (P 0.05). (2) Myelogenic monocyte knockout ADK enhanced osteoclast resorption function: The results of osteoclast bone resorption in vitro in different genotypes of mice showed that ADK f was significantly higher than that in ADK f/f group. Osteoclast lacunae area and number of osteoclasts cultured in / fLysmcre / CRE group increased significantly (P 0.01) and increased significantly (P 0.05). There was no significant difference in lacuna area and number of osteoclasts cultured in ADKf / fLysmcre /+ group compared with ADKf / fLysmcre / CRE group (P 0.05). ABT-702 intervention osteoclasts bone resorption experiment results showed that compared with 5% Et OH group, 100nM bone lacuna area increased and the number increased, there was a significant difference (P 0.01). Osteoclasts absorption function related gene RT-PCR detection results, compared with ADKf/fLysmcre/cre group, ADKf/fLysmcre/cre group MMP-9, Cts K, CTR gene expression increased, with a significant difference. There was no significant difference in the expression of c-Src gene (P 0.05) between ADKf/f Lysmcre/+ group and ADKf/f Lysmcre/cre group (P 0.05). Compared with ADKf/f Lysmcre/cre group, the expression of MMP-9, Cts-K and CTR gene in ADKf/f Lysmcre/+ group was significantly different (P 0.05). The results of ABT-702 intervention osteoclast bone resorption related gene showed that the expression of ADKf/f Lysmcre/f Lysmcre/cre group was significantly different from that of ADKf Compared with 5% Et OH group, the expression of MMP-9 (P 0.01) and Cts K (P 0.05) increased in 100N M group, but the expression of c-Src and CTR genes did not change significantly (P 0.05). 3. Bone mass of ADK mice with myelomonocyte knockout was the lightest in body weight and femoral wet weight in ADK f/smcre/cre mice, with significant difference (P 0.01). Compared with ADKf/fLysmcre/cre mice, the body weight and femoral weight of ADKf/fLysmcre/+ mice were significantly different (P In ADKf / fLysmcre /+ group, the trabecular bone structure was between the two groups, and the trabecular bone was slender and partially broken. Static parameters of bone tissue showed that ADKf / flysmcre / CRE mice were significantly different from ADKf / F mice, ADKf / flysmcre /+ mice,% Tb. Ar, Tb. Th, Tb. N,% Ob. S / BS, Ob. N / BS, Tb. Sp,% Oc. N / BS (P 0.05), respectively. Segment analysis showed that compared with the other two groups,% Ct. Ar,% Ma. Ar parameters changed significantly (P 0.01). 4. FTZ inhibited the osteoclast differentiation and absorption function at different doses. Compared with the control group, the number of osteoclasts (P 0.01), the area of bone lacunae (P 0.01) and the number of osteoclasts (P 0.01) in the Alendronate group, FTZ-H group and FTZ-M group were inhibited. The number of osteoclasts and lacunae in FTZ-L group was significantly lower than that in FTZ-L group (P 0.05), and the number of osteoclasts and lacunae in FTZ-H group (P 0.01) and FTZ-M group (P 0.05). The expression of FATc1 gene was significantly decreased (P 0.05), but the expression of c-Fos gene was not statistically significant (P 0.05). The expression of MMP-9 gene in FTZ-H group was significantly lower than that in control group (P 0.01). The expression of Cts K, c-Src and CTR gene was not significantly changed in FTZ-H group (P 0.05). [Conclusion] Cell knockout ADK mice enhanced osteoclast differentiation. 2, myelomonocyte knockout ADK mice enhanced osteoclast absorption. 3, myelomonocyte knockout ADK mice showed decreased bone mass and abnormal bone structure. 4, FTZ improved osteoporosis by inhibiting osteoclast differentiation and absorption.
【學(xué)位授予單位】：廣東藥科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R580
[Abstract]:[Objective] Compared with the control group, the weight of ADK mice was significantly decreased during the construction of ADK knockout mice. Therefore, the osteoclast (OC) differentiation and bone resorption function of ADK mice were studied by using myelomonocyte line specific knockout mice. The effect of sexual monocyte knockout ADK on bone metabolism was evaluated in vitro. The effects of Zhenzhu Tiaozhi capsule (FTZ) on osteoclast differentiation and bone resorption were evaluated under the established osteoclast culture system. ADK f/f, ADK f/flysmcre/cre, ADK f/flysmcre/+ three gene knockout mice were identified and screened. The expression of ADK protein in primary macrophages of different genotypes of mice was detected by Western blot. 2. Osteoclast differentiation and bone resorption function in vitro were detected by ADK f/f, ADK f/flysmcre/cre, ADK f/flysmcre/+ three gene knockout mice. Osteoclasts were induced by M-CSF and RANKL stimulation. Primary bone marrow mononuclear cells of wild type BL/6J mice were collected and interfered with ADK inhibitor (ABT-702), tartrate-resistant acid phosphatase specific Trap staining, bone resorption test, TRAP, NFATc1 related to bone metabolism were performed. The effects of myelomonocyte knockout ADK on osteoclast differentiation and bone resorption in vitro were evaluated by RT-PCR. 3. Bone morphometry of 4-month-old ADK f/f, ADK f/flysmcre/cre, ADK f/flysmcre/+ male mice was compared with that of 4-month-old ADK f/f, ADK f/flysmcre/+ male mice. HE staining was used to observe the pathomorphological changes of bone tissue in mice. The changes of bone mass in mice were analyzed by bone histostatic analysis system. 4. The effect of FTZ on osteoclast differentiation and bone resorption in vitro. The primary bone marrow mononuclear cells of wild type BL/6J mice were collected and added into the culture system of osteoclasts in vitro. Drug serum intervention, tartrate-resistant acid phosphatase-specific TRAP staining, bone resorption test, Bone Metabolism-Related TRAP, NFATc1, c-Fos, MMP-9, Cts K, c-Src, CTR gene RT-PCR assay were used to evaluate the effect of FTZ on osteoclast differentiation and absorption function. [Results] 1. The knockout efficiency of ADKf/smcre/cre/ADflyK was higher than that of ADKf/fLysms/+mice. ADK f / flysmcre / CRE mice had a clear band at 350 bp. ADK f / flysmcre / CRE mice had a clear band at 700 bp. ADK f / flysmcre / + mice had two bands at 350 BP and 700 bp. (2) Western blot was used to detect ADK protein level in myeloid monocytes. Compared with ADK f/f group, ADK protein expression in ADK f/fLysmcre/cre group was decreased by 69.77% (P 0.01), and ADK protein expression in ADK f/fLysmcre/+ group was decreased by 36.83% (P 0.05). 2. Osteoclast differentiation and bone resorption were increased in ADK-knockout mice. (1) Osteoclast differentiation was increased in ADK-knockout mice: no The results of osteoclasts culture in vitro showed that the number of osteoclasts in ADKf/fLysmcre/cre group was significantly higher than that in ADKf/fLysmcre/cre group (P 0.01), but there was no significant change in the number of osteoclasts in ADKf/fLysmcre/+ group (P 0.05). The results showed that the number of osteoclasts in 100nM group was higher than that in control group and 5% Et OH group (P 0.01). The expression of NFATc1 (P 0.05) gene in ADKf / fLysmcre / CRE group was higher than that in ADKf / F group by RT-PCR. There was no significant difference in the expression of OS gene (P 0.05), but there was no significant change in the above genes in ADKf / fLysmcre /+ group (P 0.05). Compared with ADKf / fLysmcre / cre group, the expression of TRAP and NFATc1 gene in ADKf / fLysmcre /+ group was significantly different (P 0.05), and there was no difference in the expression of c-Fos gene (P 0.05). The results of RT-PCR showed that ABT-702 intervened osteoclast differentiation-related genes. Compared with 5% Et OH group, the expression of TRAP and NFATc1 m RNA in 100N M group increased, with statistical difference (P 0.05); the expression of c-Fos gene had no significant difference (P 0.05). (2) Myelogenic monocyte knockout ADK enhanced osteoclast resorption function: The results of osteoclast bone resorption in vitro in different genotypes of mice showed that ADK f was significantly higher than that in ADK f/f group. Osteoclast lacunae area and number of osteoclasts cultured in / fLysmcre / CRE group increased significantly (P 0.01) and increased significantly (P 0.05). There was no significant difference in lacuna area and number of osteoclasts cultured in ADKf / fLysmcre /+ group compared with ADKf / fLysmcre / CRE group (P 0.05). ABT-702 intervention osteoclasts bone resorption experiment results showed that compared with 5% Et OH group, 100nM bone lacuna area increased and the number increased, there was a significant difference (P 0.01). Osteoclasts absorption function related gene RT-PCR detection results, compared with ADKf/fLysmcre/cre group, ADKf/fLysmcre/cre group MMP-9, Cts K, CTR gene expression increased, with a significant difference. There was no significant difference in the expression of c-Src gene (P 0.05) between ADKf/f Lysmcre/+ group and ADKf/f Lysmcre/cre group (P 0.05). Compared with ADKf/f Lysmcre/cre group, the expression of MMP-9, Cts-K and CTR gene in ADKf/f Lysmcre/+ group was significantly different (P 0.05). The results of ABT-702 intervention osteoclast bone resorption related gene showed that the expression of ADKf/f Lysmcre/f Lysmcre/cre group was significantly different from that of ADKf Compared with 5% Et OH group, the expression of MMP-9 (P 0.01) and Cts K (P 0.05) increased in 100N M group, but the expression of c-Src and CTR genes did not change significantly (P 0.05). 3. Bone mass of ADK mice with myelomonocyte knockout was the lightest in body weight and femoral wet weight in ADK f/smcre/cre mice, with significant difference (P 0.01). Compared with ADKf/fLysmcre/cre mice, the body weight and femoral weight of ADKf/fLysmcre/+ mice were significantly different (P In ADKf / fLysmcre /+ group, the trabecular bone structure was between the two groups, and the trabecular bone was slender and partially broken. Static parameters of bone tissue showed that ADKf / flysmcre / CRE mice were significantly different from ADKf / F mice, ADKf / flysmcre /+ mice,% Tb. Ar, Tb. Th, Tb. N,% Ob. S / BS, Ob. N / BS, Tb. Sp,% Oc. N / BS (P 0.05), respectively. Segment analysis showed that compared with the other two groups,% Ct. Ar,% Ma. Ar parameters changed significantly (P 0.01). 4. FTZ inhibited the osteoclast differentiation and absorption function at different doses. Compared with the control group, the number of osteoclasts (P 0.01), the area of bone lacunae (P 0.01) and the number of osteoclasts (P 0.01) in the Alendronate group, FTZ-H group and FTZ-M group were inhibited. The number of osteoclasts and lacunae in FTZ-L group was significantly lower than that in FTZ-L group (P 0.05), and the number of osteoclasts and lacunae in FTZ-H group (P 0.01) and FTZ-M group (P 0.05). The expression of FATc1 gene was significantly decreased (P 0.05), but the expression of c-Fos gene was not statistically significant (P 0.05). The expression of MMP-9 gene in FTZ-H group was significantly lower than that in control group (P 0.01). The expression of Cts K, c-Src and CTR gene was not significantly changed in FTZ-H group (P 0.05). [Conclusion] Cell knockout ADK mice enhanced osteoclast differentiation. 2, myelomonocyte knockout ADK mice enhanced osteoclast absorption. 3, myelomonocyte knockout ADK mice showed decreased bone mass and abnormal bone structure. 4, FTZ improved osteoporosis by inhibiting osteoclast differentiation and absorption.
【學(xué)位授予單位】：廣東藥科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R580
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 林澤苗;鐘佳賢;賈歡歡;吳玉娥;陳s，
本文編號(hào)：2241812
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