84年鼠女哪年财运最旺,857comvvv色九欧美激情|85PO_87国产精品欲av国产av资源

[1]王吉利,萬(wàn)雷,張志海,等.抑制Dickkopf-1和Sclerostin表達(dá)對(duì)人成骨肉瘤細(xì)胞MG63骨代謝調(diào)節(jié)相關(guān)蛋白表達(dá)水平的影響[J].中醫(yī)正骨,2016,28(09):13-18.
　WANG Jili,WAN Lei,ZHANG Zhihai,et al.Effect of suppression of Dickkopf-1 and Sclerostin expression on bone metabolism regulatory protein expression levels in human osteosarcoma MG63 cells[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(09):13-18.
點(diǎn)擊復(fù)制

抑制Dickkopf-1和Sclerostin表達(dá)對(duì)人成骨肉瘤細(xì)胞MG63骨代謝調(diào)節(jié)相關(guān)蛋白表達(dá)水平的影響()
分享到：

《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第28卷
期數(shù):
2016年09期
頁(yè)碼:
13-18
欄目:
基礎(chǔ)研究
出版日期:
2016-09-20

文章信息/Info

Title:
Effect of suppression of Dickkopf-1 and Sclerostin expression on bone metabolism regulatory protein expression levels in human osteosarcoma MG63 cells
作者:
王吉利1; 萬(wàn)雷1; 張志海1; 黃宏興1; 黃紅2; 肖本浩1; 魏合偉1; 曾國(guó)勇3
1.廣州中醫(yī)藥大學(xué)第三附屬醫(yī)院,廣東 廣州 510240; 2.廣州中醫(yī)藥大學(xué),廣東 廣州 510006; 3.廣東省茂名市中醫(yī)院,廣東 茂名 525000
Author(s):
WANG Jili1; WAN Lei1; ZHANG Zhihai1; HUANG Hongxing1; HUANG Hong2; XIAO Benhao1; WEI Hewei1; ZENG Guoyong3
1.The Third Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510240,Guangdong,China2.Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006,Guangdong,China3.Maoming Hospital of Traditional Chinese Medici
關(guān)鍵詞:
骨質(zhì)疏松;  基因沉默;  Dickkopf-1;  Sclerostin;  腺病毒;  MG63細(xì)胞
Keywords:
osteoporosis;  gene silencing;  Dickkopf-1;  Serostin;  adenovirus;  MG63 cells
摘要:
目的:觀察抑制Dickkopf-1和Sclerostin表達(dá)對(duì)人成骨肉瘤細(xì)胞MG63骨代謝調(diào)節(jié)相關(guān)蛋白表達(dá)水平的影響。方法:培養(yǎng)MG63細(xì)胞,構(gòu)建沉默Dickkopf-1重組腺病毒載體和沉默Sclerostin重組腺病毒載體。將培養(yǎng)好的MG63細(xì)胞(每孔2×105個(gè))分為4組,Scr組以Scr腺病毒轉(zhuǎn)染、Dickkopf-1組以沉默Dickkopf-1重組腺病毒載體轉(zhuǎn)染、Sclerostin組以沉默Sclerostin重組腺病毒載體轉(zhuǎn)染、Dickkopf-1+Sclerostin組以沉默Dickkopf-1重組腺病毒載體和沉默Sclerostin重組腺病毒載體共同轉(zhuǎn)染。各組MG63細(xì)胞轉(zhuǎn)染重組腺病毒48 h后以Western Blot法測(cè)定各組細(xì)胞中骨保護(hù)素(osteoprotegerin,OPG)、低密度脂蛋白受體相關(guān)蛋白-5(low density lipoprotein receptor-related protein-5,Lrp-5)、骨形態(tài)發(fā)生蛋白-2(bone morphogenetic protein-2,BMP-2)、成纖維細(xì)胞生長(zhǎng)因子-2(fibroblast growth factor-2,FGF-2)、Runt相關(guān)轉(zhuǎn)錄因子-2(Runt-related transcription factor-2,Runx-2)、結(jié)締組織生長(zhǎng)因子(connective tissue growth factor,CTGF)、骨橋蛋白(osteopontin,OPN)、腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)等骨代謝調(diào)節(jié)相關(guān)蛋白的表達(dá)水平。結(jié)果:Dickkopf-1組的OPG表達(dá)量與Scr組、Sclerostin組比較,組間差異均無(wú)統(tǒng)計(jì)學(xué)意義(P=0.050,P=0.196); Sclerostin組、Dickkopf-1+Sclerostin組的OPG表達(dá)量均高于Scr組(P=0.010,P=0.000); Dickkopf-1組和Sclerostin組的OPG表達(dá)量均低于Dickkopf-1+Sclerostin組(P=0.000,P=0.000)。Dickkopf-1組、Sclerostin組、Dickkopf-1+Sclerostin組的Lrp-5表達(dá)量均高于Scr組(P=0.012,P=0.010,P=0.000); Dickkopf-1組的Lrp-5表達(dá)量與Sclerostin組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.119); Dickkopf-1組和Sclerostin組的Lrp-5表達(dá)量均低于Dickkopf-1+Sclerostin組(P=0.000,P=0.000)。Dickkopf-1組的BMP-2表達(dá)量與Scr組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.185); Dickkopf-1組的BMP-2表達(dá)量低于Sclerostin組(P=0.037); Dickkopf-1組和Sclerostin組的BMP-2表達(dá)量均低于Dickkopf-1+Sclerostin組(P=0.000,P=0.000)。Dickkopf-1組、Sclerostin組、Dickkopf-1+Sclerostin組的FGF-2表達(dá)量均高于Scr組(P=0.010,P=0.000,P=0.000); Dickkopf-1組的FGF-2表達(dá)量低于Sclerostin組和Dickkopf-1+Sclerostin組(P=0.000,P=0.000); Sclerostin組的FGF-2表達(dá)量低于Dickkopf-1+Sclerostin組(P=0.000)。Dickkopf-1組、Sclerostin組、Dickkopf-1+Sclerostin組的Runx-2表達(dá)量均高于Scr組(P=0.000,P=0.000,P=0.000); Dickkopf-1組的Runx-2表達(dá)量低于Sclerostin組和Dickkopf-1+Sclerostin組(P=0.000,P=0.000); Sclerostin組的Runx-2表達(dá)量低于Dickkopf-1+Sclerostin組(P=0.000)。Dickkopf-1組、Sclerostin組、Dickkopf-1+Sclerostin組的CTGF表達(dá)量均高于Scr組(P=0.010,P=0.000,P=0.010); Dickkopf-1組的CTGF表達(dá)量與Sclerostin組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.080); Dickkopf-1組和Sclerostin組的CTGF表達(dá)量均低于Dickkopf-1+Sclerostin組(P=0.000,P=0.004)。Dickkopf-1組、Sclerostin組、Dickkopf-1+Sclerostin組的OPN表達(dá)量均高于Scr組(P=0.002,P=0.000,P=0.000); Dickkopf-1組和Dickkopf-1+Sclerostin組的OPN表達(dá)量與Sclerostin組比較,差異均無(wú)統(tǒng)計(jì)學(xué)意義(P=0.050,P=0.170); Dickkopf-1組的OPN表達(dá)量低于Dickkopf-1+Sclerostin組(P=0.000)。Dickkopf-1組、Sclerostin組、Dickkopf-1+Sclerostin組的TNF-α表達(dá)量均低于Scr組(P=0.000,P=0.000,P=0.000); Dickkopf-1組的TNF-α表達(dá)量高于Sclerostin組和Dickkopf-1+Sclerostin組(P=0.000,P=0.000); Sclerostin組和Dickkopf-1+Sclerostin組的TNF-α表達(dá)量比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.125)。結(jié)論:抑制MG63細(xì)胞Dickkopf-1和Sclerostin表達(dá),均能上調(diào)具有骨形成和雙向調(diào)節(jié)作用的骨代謝調(diào)節(jié)相關(guān)蛋白表達(dá)水平、下調(diào)具有骨吸收作用的骨代謝調(diào)節(jié)相關(guān)蛋白表達(dá)水平,其中抑制Sclerostin表達(dá)對(duì)骨代謝調(diào)節(jié)相關(guān)蛋白表達(dá)水平的影響強(qiáng)于抑制Dickkopf-1表達(dá),而且抑制Dickkopf-1和Sclerostin表達(dá)在調(diào)節(jié)骨代謝調(diào)節(jié)-----------------------------------------------
Abstract:
Objective:To observe the effect of suppression of Dickkopf-1 and Sclerostin expression on bone metabolism regulatory protein expression levels in human osteosarcoma MG63 cells.Methods:The MG63 cells were cultured and Dickkopf-1 silencing recombinant adenovirus vectors and Sclerostin silencing recombinant adenovirus vectors were constructed.The MG63 cells were divided into 4 groups,two hundred thousand cells in one hole,and were transfected with Scr adenovirus(Scr group),Dickkopf-1 silencing recombinant adenovirus vectors(Dickkopf-1 group),Sclerostin silencing recombinant adenovirus vectors(Sclerostin group)and Dickkopf-1 silencing recombinant adenovirus vectors combined with Sclerostin silencing recombinant adenovirus vectors(Dickkopf-1-Sclerostin group)respectively.At 48 hours after recombinant adenovirus transfection,Western Blot was used to measure the expression levels of bone metabolism regulatory protein,including osteoprotegerin(OPG),low density lipoprotein receptor-related protein-5(Lrp-5),bone morphogenetic protein-2(BMP-2),fibroblast growth factor-2(FGF-2),Runt-related transcription factor-2(Runx-2),connective tissue growth factor(CTGF),osteopontin(OPN)and tumor necrosis factor-α(TNF-α).Results:There was no statistical difference in the expression of OPG between Dickkopf-1 group and Scr group and between Dickkopf-1 group and Sclerostin group(P=0.050,P=0.196).The expression of OPG was higher in Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.010,P=0.000),and was lower in Dickkopf-1 group and Sclerostin group compared to Dickkopf-1-Sclerostin group(P=0.000,P=0.000).The expression of Lrp-5 was higher in Dickkopf-1 group,Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.012,P=0.010,P=0.000).There was no statistical differences in the expression of Lrp-5 between Dickkopf-1 group and Sclerostin group(P=0.119).The expression of Lrp-5 was lower in Dickkopf-1 group and Sclerostin group compared to Dickkopf-1-Sclerostin group(P=0.000,P=0.000).There was no statistical differences in the expression of BMP-2 between Dickkopf-1 group and Scr group(P=0.185).The expression of BMP-2 was lower in Dickkopf-1 group compared to Sclerostin group(P=0.037),and was lower in Dickkopf-1 group and Sclerostin group compared to Dickkopf-1-Sclerostin group(P=0.000,P=0.000).The expression of FGF-2 was higher in Dickkopf-1 group,Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.010,P=0.000,P=0.000),and was lower in Dickkopf-1 group compared to Sclerostin group and Dickkopf-1-Sclerostin group(P=0.000,P=0.000).The expression of FGF-2 was lower in Sclerostin group compared to Dickkopf-1-Sclerostin group(P=0.000).The expression of Runx-2 was higher in Dickkopf-1 group,Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.000,P=0.000,P=0.000).The expression of Runx-2 was lower in Dickkopf-1 group compared to Sclerostin group and Dickkopf-1-Sclerostin group(P=0.000,P=0.000),and was lower in Sclerostin group compared to Dickkopf-1-Sclerostin group(P=0.000).The expression of CTGF was higher in Dickkopf-1 group,Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.010,P=0.000,P=0.010).There was no statistical differences in the expression of CTGF between Dickkopf-1 group and Sclerostin group(P=0.080).The expression of CTGF was lower in Dickkopf-1 group and Sclerostin group compared to Dickkopf-1-Sclerostin group(P=0.000,P=0.004).The expression of OPN was higher in Dickkopf-1 group,Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.002,P=0.000,P=0.000).There was no statistical differences in the expression of OPN between Dickkopf-1 group and Sclerostin group and between Dickkopf-1-Sclerostin group and Sclerostin group(P=0.050,P=0.170).The expression of OPN was lower in Dickkopf-1 group compared to Dickkopf-1-Sclerostin group(P=0.000).The expression of TNF-α was lower in Dickkopf-1 group,Sclerostin group and Dickkopf-1-Sclerostin group compared to Scr group(P=0.000,P=0.000,P=0.000),and was higher in Dickkopf-1 group compared to Sclerostin group and Dickkopf-1-Sclerostin group(P=0.000,P=0.000).There was no statistical differences in the expression of TNF-α between Sclerostin group and Dickkopf-1-Sclerostin group(P=0.125).Conclusion:By suppressing the expression of Dickkopf-1 and Sclerostin in MG63 cells,the expression levels of bone metabolism regulatory proteins which has accelerating effects on bone formation and which has bidirectional regulatory effects on bone metabolism can be upregulated,and the expression levels of bone metabolism regulatory proteins which has accelerating effects on bone resorption can be downregulated.The expression levels of bone metabolism regulatory proteins are influenced more by suppression of Sclerostin expression than suppression of Dickkopf-1 expression,and the suppression of expression of Dickkopf-1 and Sclerostin has a synergistic effect on regulating the expression levels of bone metabolism regulatory proteins.

參考文獻(xiàn)/References:

[1] Ke HZ,Richards WG,Li XD,et al.Sclerostin and dickkopf-1 as therapeutic targets in bone diseases[J].Endocr Rev,2012,33(5):747-783. [2] Macdonald BT,Joiner DM,Oyserman SM,et al.Bone mass is inversely proportional to Dkk1 levels in mice[J].Bone,2007,41(3):331-339. [3] Wang FS,Ko JY,Yeh DW,et al.Modulation of dickkopf-1 attenuates glucocorticoid induction of osteoblast apoptosis,adipocytic differentiation,and bone mass loss[J].Endocrinology,2008,149(4):1793-1801. [4] Ellies DL,Viviano B,Mccarthy J,et al.Bone density ligand, Sclerostin, directly interacts with LRP5 but not LRP5G171V to modulate Wnt activity[J].J Bone Miner Res,2006,21(11):1738-1749. [5] Ryan ZC,Ketha H,Mcnulty MS,et al.Sclerostin alters serum vitamin D metabolite and fibroblast growth factor 23 concentrations and the urinary excretion of Calcium[J].Proc Natl Acad Sci U S A,2013,110(15):6199-6204. [6] 萬(wàn)雷,黃宏興,黃紅,等.過(guò)表達(dá)DKK1,Sost重組腺病毒載體的構(gòu)建及其對(duì)MG63細(xì)胞和相關(guān)蛋白的影響研究[J].廣州中醫(yī)藥大學(xué)學(xué)報(bào),2016,33(4):578-584. [7] Rodda SJ,Mcmahon AP.Distinct roles for Hedgehog and canonical Wnt signaling in specification,differentiation and maintenance of osteoblast progenitors[J].Development,2006,133(16):3231-3244. [8] Rawadi G,Vayssière B,Dunn F,et al.BMP-2 controls alkaline phosphatase expression and osteoblast mineralization by a Wnt autocrine loop[J].J Bone Miner Res,2003,18(10):1842-1853. [9] LEUNGD.Adenoviral vectors[J].Meducator,2012,1(2):6. [10] Boyce BF,Hughes DE,Wright KR,et al.Recent advances in bone biology provide insight into the pathogenesis of bone diseases[J].Lab Invest,1999,79(2):83-94. [11] 許勇,王鑄鋼,楊樺,等.骨保護(hù)素(OPG)基因敲除小鼠發(fā)生高轉(zhuǎn)換型骨質(zhì)疏松和動(dòng)脈鈣化[J].生物化學(xué)與生物物理進(jìn)展,2007,34(003):260-266. [12] Yadav VK,Ryu JH,Suda N,et al.Lrp5 controls bone formation by inhibiting serotonin synthesis in the duodenum[J].Cell,2008,135(5):825-837. [13] Zhao M,Ko SY,Liu JH,et al.Inhibition of microtubule assembly in osteoblasts stimulates bone morphogenetic protein 2 expression and bone formation through transcription factor Gli2[J].Mol Cell Biol,2009,29(5):1291-1305. [14] Zoricic S,Maric I,Bobinac D,et al.Expression of bone morphogenetic proteins and cartilage-derived morphogenetic proteins during osteophyte formation in humans[J].J Anat,2003,202(Pt 3):269-277. [15] Mao B,Wu W,Davidson G,et al.Kremen proteins are Dickkopf receptors that regulate Wnt/beta-catenin signalling[J].Nature,2002,417(6889):664-667. [16] Mao B,Wu W,Li Y,et al.LDL-receptor-related protein 6 is a receptor for Dickkopf proteins[J].Nature,2001,411(6835):321-325. [17] Hess K,Ushmorov A,Fiedler J,et al.TNFalpha promotes osteogenic differentiation of human mesenchymal stem cells by triggering the NF-kappaB signaling pathway[J].Bone,2009,45(2):367-376. [18] Ding J,Ghali O,Lencel P,et al.TNF-alpha and IL-1beta inhibit RUNX2 and collagen expression but increase alkaline phosphatase activity and mineralization in human mesenchymal stem cells[J].Life Sci,2009,84(15/16):499-504. [19] 張麗,姜麗燕,張良巖,等.CTGF 對(duì)破骨前體細(xì)胞 RAW264.7增殖及碳酸酐酶Ⅱ分化的影響[J].中國(guó)骨質(zhì)疏松雜志,2014,202(2):133-136. [20] 王增田,王心,楊靜,等.骨橋蛋白影響破骨細(xì)胞,成骨細(xì)胞參與骨重塑過(guò)程的研究進(jìn)展[J].武警后勤學(xué)院學(xué)報(bào):醫(yī)學(xué)版,2014,23(12):1062.

相似文獻(xiàn)/References:

[1]劉磊,萬(wàn)春友,全先輝,等.鎖定鋼板內(nèi)固定結(jié)合肱骨頭植骨重建治療 肱骨近端Neer三、四部分骨折[J].中醫(yī)正骨,2015,27(10):52.
[2]徐偉鋒,葉健,吳連國(guó).強(qiáng)骨飲對(duì)骨質(zhì)疏松性股骨頸骨折患者全髖關(guān)節(jié)置換術(shù)后 血清骨代謝生化指標(biāo)和骨密度的影響[J].中醫(yī)正骨,2015,27(02):12.
　XU Weifeng,YE Jian,WU Lianguo.Effect of Qianggu Yin(強(qiáng)骨飲,QGY)on serum bone metabolism indexes and bone density after total hip arthroplasty in patients with osteoporotic femoral neck fractures[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(09):12.
[3]龔江浩.股骨近端防旋髓內(nèi)釘聯(lián)合抗骨質(zhì)疏松藥物治療 不穩(wěn)定型老年股骨轉(zhuǎn)子間骨折的臨床觀察[J].中醫(yī)正骨,2015,27(04):29.
　GONG Jianghao.A combination therapy of proximal femoral nail antirotation and anti-osteoporotic drugs for the treatment of unstable femoral intertrochanteric fractures in the aged[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(09):29.
[4]李冠慧,李燦東,李西海,等.雌激素調(diào)控絕經(jīng)后骨質(zhì)疏松癥骨吸收-骨形成 耦聯(lián)失衡的機(jī)制[J].中醫(yī)正骨,2016,28(02):36.
[5]費(fèi)國(guó)芳,費(fèi)紅良,王金法.兩種內(nèi)固定方法治療老年不穩(wěn)定性股骨轉(zhuǎn)子間骨折的對(duì)比研究[J].中醫(yī)正骨,2016,28(05):9.
　FEI Guofang,FEI Hongliang,WANG Jinfa.Comparison of two kinds of internal fixation for treatment of unstable femoral intertrochanteric fractures in the aged[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(09):9.
[6]陳思凱,邢金明.骨水泥強(qiáng)化椎弓根螺釘內(nèi)固定治療老年胸腰椎壓縮性骨折[J].中醫(yī)正骨,2016,28(05):35.
[7]孫鵬,譚磊,閻乾,等.人工肱骨頭置換術(shù)治療老年肱骨近端Neer三、四部分骨折[J].中醫(yī)正骨,2016,28(05):43.
[8]王博,吳鵬,史曉林.強(qiáng)骨飲對(duì)去卵巢骨質(zhì)疏松大鼠骨顯微結(jié)構(gòu)的影響[J].中醫(yī)正骨,2016,28(07):6.
　WANG Bo,WU Peng,SHI Xiaolin.A retrospective trial of postural reduction combined with percutaneous vertebroplasty versus percutaneous kyphoplasty for treatment of osteoporotic vertebral compression fractures[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(09):6.
[9]馮建軍,鄒浩,朱海燕,等.槲皮素對(duì)去卵巢大鼠血清骨鈣素和股骨Ⅰ型膠原蛋白水平的影響[J].中醫(yī)正骨,2016,28(07):10.
　FENG Jianjun,ZOU Hao,ZHU Haiyan,et al.Leverage bone grafting and pedicle screw internal fixation through posterior approach for treatment of thoracolumbar fractures[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(09):10.
[10]張國(guó)文.肱骨近端鎖定接骨板內(nèi)固定治療肱骨近端骨折[J].中醫(yī)正骨,2016,28(08):50.

備注/Memo

備注/Memo:
2016-07-14收稿 2016-08-09修回
基金項(xiàng)目:國(guó)家自然科學(xué)基金項(xiàng)目(81302991,81373653); 廣東省自然科學(xué)基金項(xiàng)目(S2013040016828,2014A030310127); 廣東省科技計(jì)劃項(xiàng)目(2013B021800058); 廣州中醫(yī)藥大學(xué)青年英才項(xiàng)目(QNYC20120119); 2014廣東省“優(yōu)青計(jì)劃”項(xiàng)目(yq2014041)
萬(wàn)雷 E-mail:[email protected]

更新日期/Last Update: 1900-01-01