基于牵张成骨技术比较两种补肾法在成血管-成骨耦联机制中的作用差异
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摘要
目的:观察两种补肾法在牵张成骨(DO)中对血管生成及新骨生成的影响,评估成骨效能,并对作用机制进行探讨。方法:50只SD大鼠随机分为5组:补肾阳组、补肾阴组、西药组、模型组和空白组,每组10只。构建胫骨DO模型,给予补肾阳法和补肾阴法干预,4周和8周后分别行X线、HE和Masson染色评估成骨情况,血管造影观察牵张区血管形成,ELISA检测骨髓中VEGF、Ang-2、Runx-2和BMP-2含量水平,qPCR检测骨中对应因子的mRNA表达。结果:X线、HE和Masson染色示补肾阳组和补肾阴组的成骨效果明显优于对照组与模型组,两种补肾法均可提高Lane-Sandhu、Huddleston分值(P<0.05)。与对照组与模型组比较,补肾阳、补肾阴两组可显著增加血管体积分数(P<0.05),上调VEGF、Ang-2和Runx-2、BMP-2及相应mRNA的表达(P<0.05),其中补肾阳组VEGF、Ang-2及其mRNA表达最高,而补肾阴组Runx-2、BMP-2及其mRNA表达最高。结论:补肾阳法和补肾阴法均可上调DO中成血管-成骨因子表达,促进血管生成和新骨形成,从而改善成骨效能、促进骨修复,但二者的作用机制存在差异。
Objective: To observe the effects of two types of tonifying kidney on angiogenesis and new bone formation in distraction osteogenesis(DO), evaluate the osteogenesis efficacy and explore the mechanism. Methods: Fifty 10-week SD rats were randomly divided into five groups: tonifying kidney-yang group, tonifying kidney-yin group, western medicine group, model group and control group. DO models of SD rats were established, and after 4 and 8 weeks, osteogenesis was evaluated by X-ray, HE and Masson staining, respectively. Angiography was used to observe the angiogenesis in the distraction area. The levels of VEGF, Ang-2, Runx-2 and BMP-2 in bone marrow were detected by ELISA. The mRNA expressions of corresponding factors were detected by q PCR. Results: X-ray, HE and Masson staining showed that the osteogenesis effects of tonifying kidney-yang and kidney-yin were significantly better than that of model group and control group(P<0.05). Both kidney-reinforcing methods could improve the LaneSandhu and Huddleston scores(P<0.05) and up-regulate the expressions of VEGF, Ang-2 and Runx-2, BMP-2 and corresponding m RNA(P<0.05). Among them, the expressions of VEGF, Ang-2 and their mRNA were highest in the tonifying kidney-yang group,whilie the expression of Runx-2, BMP-2 and their mRNA were highest in the tonifying kidney-yin group. Conclusion: Both tonifying kidney-yang and kidney-yin could promote the expressions of angiogenic-osteogenic factors, promote angiogenesis and new bone formation, and improve the osteogenic efficacy in DO, but the mechanisms of the two methods are different.
Objective: To observe the effects of two types of tonifying kidney on angiogenesis and new bone formation in distraction osteogenesis(DO), evaluate the osteogenesis efficacy and explore the mechanism. Methods: Fifty 10-week SD rats were randomly divided into five groups: tonifying kidney-yang group, tonifying kidney-yin group, western medicine group, model group and control group. DO models of SD rats were established, and after 4 and 8 weeks, osteogenesis was evaluated by X-ray, HE and Masson staining, respectively. Angiography was used to observe the angiogenesis in the distraction area. The levels of VEGF, Ang-2, Runx-2 and BMP-2 in bone marrow were detected by ELISA. The mRNA expressions of corresponding factors were detected by q PCR. Results: X-ray, HE and Masson staining showed that the osteogenesis effects of tonifying kidney-yang and kidney-yin were significantly better than that of model group and control group(P<0.05). Both kidney-reinforcing methods could improve the LaneSandhu and Huddleston scores(P<0.05) and up-regulate the expressions of VEGF, Ang-2 and Runx-2, BMP-2 and corresponding m RNA(P<0.05). Among them, the expressions of VEGF, Ang-2 and their mRNA were highest in the tonifying kidney-yang group,whilie the expression of Runx-2, BMP-2 and their mRNA were highest in the tonifying kidney-yin group. Conclusion: Both tonifying kidney-yang and kidney-yin could promote the expressions of angiogenic-osteogenic factors, promote angiogenesis and new bone formation, and improve the osteogenic efficacy in DO, but the mechanisms of the two methods are different.
引文
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[22]陈伟健,晋大祥,谢炜星,等.Runx2基因参与骨代谢相关通路的研究进展.中国骨质疏松杂志,2018,24(4):557-560
[23]Abe E.Function of BMPs and BMP antagonists in adult bone.Ann N Y Acad Sci,2006,1068(1):41-53
[24]Zhang C,Meng C,Guan D.BMP2 and VEGF165 transfection to bone marrow stromal stem cells regulate osteogenic potential in vitro.Medicine,2018,97(5):e9787
[25]孙明,律娜,张令达,等.牵张成骨后移动骨段移位对早期血管生成的影响.安徽医科大学学报,2011,46(11):1130-1133
[2]姜自伟,曾景奇,黄枫,等.骨碎补总黄酮对大鼠胫骨牵张成骨效能的影响.中华中医药杂志,2018,33(2):661-663
[3]余翔,姜自伟,郑晓辉,等.补肾中药调控BMP-Smad信号通路促进牵张成骨的研究进展.辽宁中医杂志,2017,44(9):1991-1994
[4]李晋玉,赵学千,孙旗,等.骨碎补总黄酮的实验及临床研究概况.中国骨质疏松杂志,2018,24(10):1357-1364
[5]Maes C.Role and regulation of vascularization processes in endochondral bones.Calcif Tissue Int,2013,92(4):307-323
[6]Kusumbe A P,Ramasamy S K.Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone.Nature,2014,507(7492):323-328
[7]李悦,姜自伟,侯雪芹,等.两种补肾法治疗绝经后骨质疏松症的分子机制差异.中华中医药志,2015,30(5):1570-1574
[8]Ilizarov G A.Clinical and experimental data on bloodless lengthening of lower extremities.Eksp Khir Anesteziol,1969,14(4):27-32
[9]Bell A,Templeman D,Weinlein J C.Nonunion of the femur and tibia:An update.Orthop Clin NorthAm,2016,47(2):365-375
[10]刘凯,周诺.牵张成骨中骨形成相关信号通路的研究进展.中国矫形外科杂志,2018,26(3):230-233
[11]Sun Y,Xu J,Xu L,et al.MiR-503 promotes bone formation in distraction osteogenesis through suppressing Smurf1 Expression.Sci Rep,2017,7(1):409
[12]Ashman O,Phillips A M.Treatment of non-unions with bone defects:which option and why?Injury,2013,44(Suppl1):43-45
[13]姜自伟,曾景奇,李悦,等.骨碎补总黄酮对大鼠胫骨牵张末期BMP-2与Smad-1表达的影响.辽宁中医杂志,2018,45(1):166-168,227
[14]罗伟东,姜自伟,李悦,等.牵张应力下骨碎补总黄酮对成骨细胞增殖及相关蛋白含量的影响.新中医,2016,48(8):299-302
[15]高怡加,黄培镇,李悦,等.骨碎补总黄酮对牵张成骨过程中骨形态发生蛋白-2和转化生长因子-β1表达的影响.广州中医药大学学报,2016,33(5):679-683
[16]沈耿杨,任辉,张志达,等.龟板联合阿伦磷酸钠对激素性骨质疏松大鼠腰椎Runx2、CTSK表达的影响.中华中医药志,2017,32(5):2181-2185
[17]任辉,张志达,梁德,等.龟板改善激素性骨质疏松大鼠骨量、骨微细结构、骨生物力学和骨代谢的机制探讨.中华中医药杂志,2016,31(5):1858-1862
[18]Hu K,Olsen B R.Osteoblast-derived VEGF regulates osteoblast differentiation and bone formation during bone repair.J Clin Invest,2016,126(2):509-526
[19]Ardelt A A,McCullough L D,Korach K S,et al.Estradiol regulates angiopoietin-1 mRNA expression through estrogen receptor-alpha in a rodent experimental stroke model.Stroke,2005,36(2):337-341
[20]Yancopoulos G D,Davis S,Gale N W,et al.Vacular specific growth factors and blood vessel formation.Nature,2000,407(6081):242-248
[21]周大凯,李慧宁,马珊珊,等.Runx2基因高表达对脐血间充质干细胞成骨分化相关基因表达的影响.中国组织工程研究,2017,21(9):1444-1449
[22]陈伟健,晋大祥,谢炜星,等.Runx2基因参与骨代谢相关通路的研究进展.中国骨质疏松杂志,2018,24(4):557-560
[23]Abe E.Function of BMPs and BMP antagonists in adult bone.Ann N Y Acad Sci,2006,1068(1):41-53
[24]Zhang C,Meng C,Guan D.BMP2 and VEGF165 transfection to bone marrow stromal stem cells regulate osteogenic potential in vitro.Medicine,2018,97(5):e9787
[25]孙明,律娜,张令达,等.牵张成骨后移动骨段移位对早期血管生成的影响.安徽医科大学学报,2011,46(11):1130-1133