青藤碱对前成骨细胞增殖、分化的影响及可能的作用机制
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摘要
目的观察青藤碱对前成骨细胞增殖分化的影响并探讨可能的作用机制。方法将不同浓度的青藤碱作用于前成骨细胞并诱导后,运用MTT检测细胞活性,ALP法测定碱性磷酸酶的含量,从基因及蛋白水平检测OPG,RANKL表达。结果青藤碱对细胞增殖率无明显影响,在一定程度上可以促进成骨细胞的分化及钙化,并能上调OPG的表达,同时下调RANKL的表达。结论盐酸青藤碱具有促进骨形成功能,并可以通过OPG/RANK/RANKL信号通路促进成骨细胞的分化成熟。
Objective To investigate the effects of Sinomenine(SIN) on osteoblastic cell proliferation and differentiation,and possible mechanisms of OPG/RANK/RANKL pathway involved. Methods Cells were treated with varying doses of SIN,MTT assay,and ALP were performed to investigate cell proliferation and mineralization. The m RNA and protein expression of the OPG,RANKL,were detected using real-time reverse transcriptase and western-blot. Results SIN had no influence on viability of MC3T3E1 cells,and promoted cell proliferation and mineralization. Additionally,up-regulated OPG,down-regulated RANKL m RNA and protein expression. Conclusion SIN promoted osteoblasts metabolism and regulated OPG/RANKL expression in vitro,suggested that SIN could be a useful agent for bone tissue regeneration.
Objective To investigate the effects of Sinomenine(SIN) on osteoblastic cell proliferation and differentiation,and possible mechanisms of OPG/RANK/RANKL pathway involved. Methods Cells were treated with varying doses of SIN,MTT assay,and ALP were performed to investigate cell proliferation and mineralization. The m RNA and protein expression of the OPG,RANKL,were detected using real-time reverse transcriptase and western-blot. Results SIN had no influence on viability of MC3T3E1 cells,and promoted cell proliferation and mineralization. Additionally,up-regulated OPG,down-regulated RANKL m RNA and protein expression. Conclusion SIN promoted osteoblasts metabolism and regulated OPG/RANKL expression in vitro,suggested that SIN could be a useful agent for bone tissue regeneration.
引文
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[13]张群燕,姚茹冰,蔡辉,等.青藤碱抑制类风湿关节炎骨破坏的研究进展[J].安徽医药,2016,20(1):1-4.
[14]方庆.N-乙酰半胱氨酸、青藤碱在骨水泥颗粒诱导的破骨细胞形成及骨溶解中的作用[D].第四军医大学,2011.
[15]丁从珠.青藤碱联合甲氨蝶呤抵制类风湿关节炎骨破坏的作用及机制研究[D].南京中医药大学,2012.
[16]丁从珠,姚瑶,方芸,等.青藤碱对胶原诱导的关节炎大鼠血清OPG/RANKL、IL-17含量的影响[J].南京中医药大学学报,2012,28(4):330-333.
[17]Tang P,Xiong Q,Ge W,Zhang L.The role of Micro RNAs in Osteoclasts and Osteoporosis[J].RNA Biology,2014,11:1355-1363.
[18]THolding CA,Findlay DM,Stamenkov R,et al.The correlation of RANK,RANKL and TNFalpha expression with bone loss volume and polyethylene wear debris around hip implants[J].Biomaterials,2006;27:5212-5219.
[2]朱士龙,陈迪钊,李勇,等.青藤碱最新研究进展[J].吉林大学学报:自然科学版,2011,32(5):95-100.
[3]瑞燕,曹柳英,王文君,等.青藤碱抗炎作用机理研究[J].广州中医药大学学报,2007,24(2):141-143.
[4]姜宇懋,王丹巧.青藤碱抗肿瘤作用机制的研究进展[J].现代药物与临床,2016,31(11):1866-1870.
[5]靳涛,徐倩,陈丹娜,等.青藤碱抗肿瘤作用研究进展[J].今日健康,2016,15(11):383-384.
[6]刘刚,王辉,张先洲,等.青藤碱清除氧自由基和抗脂质过氧化作用[J].中草药,2006,37(1):84-87.
[7]陈蕊,刘昌华.正清风痛宁联合缬沙坦治疗糖尿病肾病38例临床观察[J].中医药导报,2012,18(11):56.
[8]Wu ZX,Liu D,Wan SY,et al.Sustained-release rh BMP-2increased bone mass and bone strength in an ovine model of postmenopausal osteoporosis[J].J Orthop Sci,2011,16(1):99-104.
[9]刘小惠,蔡素芬,吴英龙,等.破骨细胞和RANKL在幼年特发性关节炎中的作用机制[J].江西医药,2015,50(6):520-522.
[10]贺龙刚.青藤碱对RANKL和LPS诱导的破骨细胞的影响及作用机制研究[D].南方医科大学,2013.
[11]Lee SU,Shin HK,Min YK,et al.Accelerates osteoblast differentiation through phosphatidylinositol 3-kinase activation and bone morphogenetic protein-2 gene expression[J].Int Immunopharma col,2008,8(5):741.
[12]丁从珠,汪悦,姚瑶,等.青藤碱治疗类风湿性关节炎抑制骨破坏的研究进展[J].南京中医药大学学报,2011,27(1):98-100.
[13]张群燕,姚茹冰,蔡辉,等.青藤碱抑制类风湿关节炎骨破坏的研究进展[J].安徽医药,2016,20(1):1-4.
[14]方庆.N-乙酰半胱氨酸、青藤碱在骨水泥颗粒诱导的破骨细胞形成及骨溶解中的作用[D].第四军医大学,2011.
[15]丁从珠.青藤碱联合甲氨蝶呤抵制类风湿关节炎骨破坏的作用及机制研究[D].南京中医药大学,2012.
[16]丁从珠,姚瑶,方芸,等.青藤碱对胶原诱导的关节炎大鼠血清OPG/RANKL、IL-17含量的影响[J].南京中医药大学学报,2012,28(4):330-333.
[17]Tang P,Xiong Q,Ge W,Zhang L.The role of Micro RNAs in Osteoclasts and Osteoporosis[J].RNA Biology,2014,11:1355-1363.
[18]THolding CA,Findlay DM,Stamenkov R,et al.The correlation of RANK,RANKL and TNFalpha expression with bone loss volume and polyethylene wear debris around hip implants[J].Biomaterials,2006;27:5212-5219.