沉默ERRα对Bak1、Bcl2腺病毒转染骨细胞的影响研究

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Abstract Objective To investigate the effect of silencedEstrogen-related receptor alpha (ERRɑ) on the cell viability and related proteins of human osteoblast-like cells MG63 transfected with Bak1 and Bcl2 recombinant adenovirus vectors.Methods Bak1 and Bcl2 adenoviral vectors were constructed.MG63 cells were transfected with shBak1 and shBcl2 adenovirus vectors and were divided into empty virus group (NC control group)，silent Bak1 group，silent Bcl2 group，and silent Bak1+Bcl2 group.ERRɑ intervened in the above four groups of cells.We used MTT assay for cell proliferation，coomassie leucoprotein quantitative assay for alkaline phosphates (ALP) activity，flow cytometry for determination of calcium ion concentration，and Western blot analysis of bone morphogenetic protein-4 ( Bone morphogenetic protein-4，BMP-4)，connective tissue growth factor (CTGF)，osteopontin (OPN)，Runt-related transcription factor 2 (Runx2)，tumor Expression of tumor necrosis factor-α (TNF-α).Results a) Compared with NC control group，cell activity，ALP activity，CTGF，OPN and Runx2 were significantly increased in (Ad-shERRɑ+Ad-shBak1) group (P＜0.01)，BMP-4 (P＜0.05)，calcium ionconcentration and TNF-ɑ decreased significantly (P＜0.01)，(Ad-shERRɑ+Ad-shBcl2) group's activity，ALP activity，BMP-4，CTGF，OPN，Runx2 were significantly decreased (P＜0.01)，calcium ion concentration and TNF-ɑ increased significantly (P＜0.01)，(Ad-shERRɑ+Ad-shBak1+shBcl2) group's calcium concentration，BMP-4，CTGF，OPN，Runx2，TNF-ɑ decreased (P＜0.05)；b) Compared with the (Ad-shERRɑ+Ad-shBak1) group，(Ad-shERRɑ+Ad-shBcl2) group's and (Ad-shERRɑ+Ad-shBak1+shBcl2) group's cell activity，ALP activity，CTGF，OPN，Runx2 decreased significantly (P＜0.01)，calcium ion concentration increased significantly (P＜0.05)，and (Ad-shERRɑ+Ad-shBcl2) group significantly decreased in BMP-4 and TNF-ɑ (P＜0.01)，(Ad-shERRɑ+Ad-shBak1+shBcl2) group's TNF-ɑ was significantly increased (P＜0.01)；c) Compared with (Ad-shERRɑ+Ad-shBcl2) group，(Ad-shERRɑ+Ad-shBak1+shBcl2) group's BMP-4，CTGF，OPN increased，cell activity，calcium ion concentration and TNF-ɑ decreased (P＜0.01).Conclusion ERRɑ silencing can increase the cell viability，BMP4，CTGF，OPN，Runx2 expression，calcium ion concentration and TNF-α expression of Bak1silencing.It can also decrease the cell viability，BMP4，CTGF，OPN and Runx2 expression of Bcl2 silencing，increase calcium ion concentration and TNF-α expression.

Key words： estrogen-related receptor alpha apoptotic protein adenovirus

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	Huang Jiachun 1
	Chai Shuang 1
	Huang Hongxing 2
	et al

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Huang Jiachun 1,Chai Shuang 1,Huang Hongxing 2, et al. Effect of Silencing ERRɑ on Osteoblast Transfected with Bak1 and Bcl2 Adenovirus Vectors[J]. sygkzz, 2018, 24(9): 806-811.

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http://www.sygkzz.com/EN/ OR http://www.sygkzz.com/EN/Y2018/V24/I9/806

［1］Raisz LG.Pathogenesis of osteoporosis：concepts，conflicts，and prospects［J］.J Clin Invest 2005，115(12)：3318-3325.
［2］Mosekilde L，Vestergaard P，Rejnmark L.The pathogenesis，treatment and prevention of osteoporosis in men［J］.Drugs，2013，73(1)：15-29.
［3］Song W，Kwak HB，Lawler JM.Exercise training attenuates age-induced changes in apoptotic signaling in rat skeletal muscle［J］.Antioxidants & redox signaling，2006，8(3-4)：517-528.
［4］刘嘉眉，李颖.成骨细胞中Caspase蛋白和Bcl-2蛋白在绝经后骨质疏松症发病机理中的作用［J］.中华关节外科杂志(电子版)，2015，9(6)：82-85.
［5］杨先文，黄宏兴，万雷，等.Bcl2和Bak基因沉默对MG-63细胞凋亡和成骨活动的影响［J］.实用医学杂志，2017，33(17)：2813-2816.
［6］HUSS JM，GARBACZ WG，XIE W.Constitutive activitiesof estrogen-related receptors：Transcriptional regulation ofmetabolism by the ERR pathways in health and disease［J］.Biochim Biophys Acta，2015(1852)：1912-1927.
［7］vincent Giguère，Na Yang，Prudimar Segui，Ronald M.Evans Identification of a new class of steroid hormone receptors［J］.Nature，1988，331(6151)：91-94.
［8］Hong H，Yang L，Stallcup MR.Hormone-independent transcriptional activation and coactivator binding by novel orphan nuclear receptor ERR3［J］.J Biol Chem，1999，274(32)：22618-22626.
［9］Giguére V.To ERR in the estrogen pathway［J］.Trends in Endocrinology & Metabolism，2002，13(5)：220-225.
［10］Shoukry A，Shalaby SM，Etewa RL，et al.Association of estrogen receptor β and estrogen-related receptor α gene polymorphisms with bone mineral density in postmenopausal women［J］.Mol Cell Biochem，2015，405(1-2)：23-31.
［11］Huang T，Liu R，Fu X，et al.Aging reduces an ERRalpha-directed mitochondrial glutaminase expression suppressing glutamine anaplerosis and osteogenic differentiation of mesenchymal stem cells［J］.Stem Cells，2017，35(2)：411-424.
［12］甘国兴，莫新民，李劲平，等.壮骨止痛方药效部位的筛选及其对骨质疏松雌鼠骨代谢相关激素的影响［J］.中国实验方剂学杂志，2011，17(15)：126-130.
［13］Chipuk JE，Moldoveanu T，Llambi F，et al.The BCL-2 family reunion［J］.Molecular cell，2010，37(3)：299-310.
［14］Hardwick JM，Soane L.Multiple functions of BCL-2 family proteins［J］.Cold Spring Harbor perspectives in biology，2013，5(2)：a008722.
［15］Vervliet T，Parys JB，Bultynck G.Bcl-2 proteins and calcium signaling：complexity beneath the surface［J］.Oncogene，2016，35(39)：5079.
［16］Rong YP，Bultynck G，Aromolaran AS，et al.The BH4 domain of Bcl-2 inhibits ER calcium release and apoptosis by binding the regulatory and coupling domain of the IP3 receptor［J］.Proc Natl Acad Sci U S A，2009，106(34)：14397-14402.
［17］Kondo A，Otsuka T，Kuroyanagi G，et al.Resveratrol inhibits BMP-4-stimulated VEGF synthesis in osteoblasts：Suppression of S6 kinase［J］.Int J Mol Med，2014，33(4)：1013-1018.
［18］Gao X，Usas A，Proto JD，et al.Role of donor and host cells in muscle-derived stem cell-mediated bone repair：differentiation vs.paracrine effects［J］.FASEB J，2014，28(8)：3792-3809.
［19］Lei ZN，Liu F，Zhang LM，et al.Bcl-2 increases stroke-induced striatal neurogenesis in adult brains by inhibiting BMP-4 function via activation of β-catenin signaling［J］.Neurochem Int，2012，61(1)：34-42.
［20］张丽，姜丽燕，张良岩，等.CTGF对破骨前体细胞RAW264.7增殖及碳酸酐酶Ⅱ分化的影响［J］.中国骨质疏松杂志，2014，20(2)：133-136.
［21］Hendesi H，Barbe MF，Safadi FF，et al.Integrin mediated adhesion of osteoblasts to connective tissue growth factor(CTGF/CCN2) induces cytoskeleton reorganization and cell differentiation［J］.PloS one，2015，10(2)：e0115325.
［22］Kothari AN，Arffa ML，Chang V，et al.Osteopontin-a master regulator of epithelial-mesenchymal transition［J］.J Clin Med，2016，5(4)：39.
［23］Maruyama Z，Yoshida CA，Furuichi T，et al.Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency［J］.Dev Dyn，2007，236(7)：1876-1890.
［24］Adhami MD，Rashid H，Chen H，et al.Runx2 activity in committed osteoblasts is not essential for embryonic skeletogenesis［J］.Connective tissue research，2014，55(sup1)：102-106.
［25］David J P，Schett G.TNF and bone［J］.J Curr Dir Autoimmun，2010(11)：135-144.
［26］Liao L，Su X，Yang X，et al.TNF-α Inhibits FoxO1 by upregulating miR-705 to aggravate oxidative damage in bone marrow-derived mesenchymal stem cells during osteoporosis［J］.Stem cells，2016，34(4)：1054-1067.
［27］Cosman F，De Beur SJ，LeBoff MS，et al.Clinician's guide to prevention and treatment of osteoporosis［J］.Osteoporos Int，2014，25(10)：2359-2381.
［28］邢艳刚，张丹丹，张文艳，等.氟对小鼠 MC3T3-E 成骨细胞内Ca²⁺，NO浓度的影响［J］.山西农业科学，2017(6)：940-943.
［29］唐宏宇，董路珏，霍少川，等.组蛋白去甲基化酶JMJD2和雌激素受体相关受体α在绝经后骨质疏松症的表达［J］.中国组织工程研究，2016，20(02)：167-172.