骨康方含药血清对新生大鼠成骨细胞增殖的影响
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摘要
目的探讨骨康方含药血清对大鼠成骨细胞增殖的影响。方法选取30只健康雌性大鼠,按照随机数字表法分为空白对照组、阳性对照组和骨康组,每组10只。空白对照组用于制备空白血清,阳性对照组用于制备骨化三醇含药血清,骨康组用于制备骨康方含药血清。另选1日龄雄性大鼠5只制备成骨细胞,分别用3组血清进行培养。实验结束后比较3组成骨细胞的增殖及周期情况。结果 5%和10%含药血清培养时,阳性对照组和骨康组培养第7天时成骨细胞增值率最高(P<0.05)。20%含药血清培养时,阳性对照组和骨康组培养第5天时成骨细胞增值率最高(P<0.05)。10%含药血清培养时,骨康组对成骨细胞增殖的平均影响率较阳性对照组高(P<0.05)。不同浓度含药血清培养时,阳性对照组和骨康组处于G0/G1期比例低于空白对照组,而S期和G2/M期比例以及PI均高于空白对照组(P<0.05);且骨康组较阳性对照组变化更为明显(P<0.05)。阳性对照组和骨康组在20%含药血清培养时处于G0/G1期比例均最低,而S期和G2/M期比例以及PI最高(P<0.05)。结论骨康方含药血清对成骨细胞的增殖具有促进作用,且高浓度作用更为显著。
Objective To investigate effect of Gukang containing serum on proliferation of osteoblasts in neonatal rats. Methods A total of 30 healthy neonatal female rats were randomly divided into blank control group, positive control group and Gukang group(n=10 for each group). Blank control group was used to prepare blank serum, and positive control group was used to prepare calcitriol containing serum, while Gukang group was used to prepare Gukang containing serum. Another 5 male rats of 1-day-old were selected to prepare osteoblasts and cultured with serum samples in three groups. At the end of the experiment, proliferation and cycle conditions of osteocytes in three groups were compared. Results Proliferation rates of osteoblasts in positive control and Gukang groups were the highest on the 7~(th) d of cultivation when cultured by 5% and 10% drug-containing serum(P<0.05). Proliferation rates of osteoblasts in positive control and Gukang groups were the highest on the 5~(th) d of cultivation when cultured by 20% drug-containing serum(P<0.05). Average effect of Gukang on osteoblast proliferation was higher than that in positive control group when cultured by 10% drug-containing serum(P<0.05). In positive control and Gukang groups, percentages of osteocytes in G0/G1 phase were lower, while percentages in S phase, G2/M phase and PI were higher than those in blank control group when cultured by different drug-containing serums(P<0.05), and the changes of osteocytes in Gukang group were more obvious than those in positive control group(P<0.05). In positive control and Gukang groups, percentages in G0/G1 phase were the lowest, while percentages of S phase, G2/M phase and PI were the highest when cultured by 20% drug-containing serum(P<0.05). Conclusion Gukang containing serum may promote proliferation of osteoblasts, and the effect is more obvious when cultured by high concentration.
Objective To investigate effect of Gukang containing serum on proliferation of osteoblasts in neonatal rats. Methods A total of 30 healthy neonatal female rats were randomly divided into blank control group, positive control group and Gukang group(n=10 for each group). Blank control group was used to prepare blank serum, and positive control group was used to prepare calcitriol containing serum, while Gukang group was used to prepare Gukang containing serum. Another 5 male rats of 1-day-old were selected to prepare osteoblasts and cultured with serum samples in three groups. At the end of the experiment, proliferation and cycle conditions of osteocytes in three groups were compared. Results Proliferation rates of osteoblasts in positive control and Gukang groups were the highest on the 7~(th) d of cultivation when cultured by 5% and 10% drug-containing serum(P<0.05). Proliferation rates of osteoblasts in positive control and Gukang groups were the highest on the 5~(th) d of cultivation when cultured by 20% drug-containing serum(P<0.05). Average effect of Gukang on osteoblast proliferation was higher than that in positive control group when cultured by 10% drug-containing serum(P<0.05). In positive control and Gukang groups, percentages of osteocytes in G0/G1 phase were lower, while percentages in S phase, G2/M phase and PI were higher than those in blank control group when cultured by different drug-containing serums(P<0.05), and the changes of osteocytes in Gukang group were more obvious than those in positive control group(P<0.05). In positive control and Gukang groups, percentages in G0/G1 phase were the lowest, while percentages of S phase, G2/M phase and PI were the highest when cultured by 20% drug-containing serum(P<0.05). Conclusion Gukang containing serum may promote proliferation of osteoblasts, and the effect is more obvious when cultured by high concentration.
引文
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[2] 张淼,玉山江·艾克木,屈小青,等.糖尿病肾病合并骨质疏松症的发病机制及临床治疗[J].甘肃医药,2016,35(1):18-20.
[3] 曲方园,陈玲,付志丹,等.运用中医肾主骨的理论治疗膝骨性关节炎临床研究[J].中国伤残医学,2016,24(10):120-121.
[4] 杨桂莲,陈明.从现代医学角度认识中医“肾主骨”之理论内涵[J].内蒙古中医药,2016,35(17):159.
[5] 张冬生.骨康方加减治疗老年人骨质疏松症疗效评价[J].养生保健指南,2016,20(24):7,25.
[6] 张毅.分析骨康方联合手法复位治疗老年肱骨外科颈骨折的临床疗效[J].医药前沿,2017,7(16):145-146.
[7] 刘炎芬,崔邦胜.骨康方治疗老年性骨质疏松症疗效观察[J].广西中医药大学学报,2017,20(2):34-35.
[8] 殷玉琨,李杰,宋洪丽,等.益肾骨康方促进小鼠体外成骨作用的研究[J].中华中医药学刊,2017,35(2):451-453.
[9] 林一峰,魏合伟,黄宏兴,等.骨康含药血清对新生大鼠成骨细胞增殖影响的研究[J].中医药学刊,2003,21(7):1116-1117.
[10] 庄洪,魏合伟,林一峰,等.中药骨康含药血清中类雌二醇样物质含量的测定[J].中医正骨,2003,15(11):14-15.
[11] 林一峰,刘庆思.骨康含药血清对成骨细胞PDGF-AA表达影响的实验研究[J].中国骨质疏松杂志,2006,12(5):459-461.
[12] 魏合伟,李钊,张志海,等.骨康含药血清对新生大鼠成骨细胞碱性磷酸酶活性的影响[J].中医药学刊,2005,23(1):77-78.
[13] 魏合伟,李钊,庄洪,等.中药骨康含药血清对TNF-α致大鼠成骨细胞凋亡的影响[J].中医正骨,2005,17(3):6-8.
[14] 万雷,黄宏兴,柴生颋,等.骨康含药血清对细胞共育体系中OPG、RANKL的影响[J].辽宁中医药大学学报,2012,14(8):78-80.
[15] 赵可伟,唐菊英,肖林,等.中药骨康含药血清培养脂肪干细胞移植促进骨质疏松性骨折愈合过程中cbfa1和OCN的表达变化[J].中国骨质疏松杂志,2013,19(6):561-567.
[16] 赵可伟,潘君贻,肖林,等.中药骨康含药血清培养脂肪干细胞移植促进骨质疏松性骨折愈合的研究[J].热带医学杂志,2013,13(2):183-187,229.
[17] 宋瑄,林楠,李伟.鲑鱼降钙素鼻喷剂治疗女性围绝经期骨质疏松症临床疗效及安全性研究[J].中华保健医学杂志,2017,19(6):532-533.
[18] 王斌,罗毅文,胡年宏.骨康方含药血清诱导大鼠骨髓基质细胞向成骨细胞方向分化的实验研究[J].中国中医骨伤科杂志,2007,15(11):32-36.
[19] 余文珍,施红.复方中药含药血清研究现状[J].中医药通报,2003,2(2):113-116.
[20] 邵敏,赵静,王斌.不同中药含药血清对体外培养成骨细胞影响的实验研究[J].中国中医骨伤科杂志,2006,14(5):19-21.
[21] 邵敏,庄洪,赵静,等.含药血清对成骨细胞矿化结节及Ⅰ型胶原形成的影响[J].中医正骨,2003,15(4):3-4.
[22] 熊学华,余克强,刘庆思.中药骨康对去势大鼠骨吸收与骨形成影响的实验研究[J].现代康复,2001,5(11B):111-112.
[23] 李颖,黄宏兴,庄洪,等.中药骨康对去势大鼠骨质疏松作用机制的实验研究[J].中华中医药杂志,2009,24(2):160-163.
[24] 熊学华,冯新送,刘庆思.中药骨康对去势大鼠血清降钙素及骨生物力学影响的实验研究[J].中国中医骨伤科杂志,2005,13(3):7-9.
[25] Gu C, Fu L, Yuan X, et al. Promoting effect of pinostrobin on the proliferation, differentiation, and mineralization of murine pre-osteoblastic MC3T3-E1 cells[J].Molecules, 2017,22(10):1735.
[26] 万雷,黄宏兴,刘庆思,等.骨康对绝经后骨质疏松症患者骨密度、血清E2、SOD、MDA的影响[J].医药前沿,2012,2(14):93-94.
[2] 张淼,玉山江·艾克木,屈小青,等.糖尿病肾病合并骨质疏松症的发病机制及临床治疗[J].甘肃医药,2016,35(1):18-20.
[3] 曲方园,陈玲,付志丹,等.运用中医肾主骨的理论治疗膝骨性关节炎临床研究[J].中国伤残医学,2016,24(10):120-121.
[4] 杨桂莲,陈明.从现代医学角度认识中医“肾主骨”之理论内涵[J].内蒙古中医药,2016,35(17):159.
[5] 张冬生.骨康方加减治疗老年人骨质疏松症疗效评价[J].养生保健指南,2016,20(24):7,25.
[6] 张毅.分析骨康方联合手法复位治疗老年肱骨外科颈骨折的临床疗效[J].医药前沿,2017,7(16):145-146.
[7] 刘炎芬,崔邦胜.骨康方治疗老年性骨质疏松症疗效观察[J].广西中医药大学学报,2017,20(2):34-35.
[8] 殷玉琨,李杰,宋洪丽,等.益肾骨康方促进小鼠体外成骨作用的研究[J].中华中医药学刊,2017,35(2):451-453.
[9] 林一峰,魏合伟,黄宏兴,等.骨康含药血清对新生大鼠成骨细胞增殖影响的研究[J].中医药学刊,2003,21(7):1116-1117.
[10] 庄洪,魏合伟,林一峰,等.中药骨康含药血清中类雌二醇样物质含量的测定[J].中医正骨,2003,15(11):14-15.
[11] 林一峰,刘庆思.骨康含药血清对成骨细胞PDGF-AA表达影响的实验研究[J].中国骨质疏松杂志,2006,12(5):459-461.
[12] 魏合伟,李钊,张志海,等.骨康含药血清对新生大鼠成骨细胞碱性磷酸酶活性的影响[J].中医药学刊,2005,23(1):77-78.
[13] 魏合伟,李钊,庄洪,等.中药骨康含药血清对TNF-α致大鼠成骨细胞凋亡的影响[J].中医正骨,2005,17(3):6-8.
[14] 万雷,黄宏兴,柴生颋,等.骨康含药血清对细胞共育体系中OPG、RANKL的影响[J].辽宁中医药大学学报,2012,14(8):78-80.
[15] 赵可伟,唐菊英,肖林,等.中药骨康含药血清培养脂肪干细胞移植促进骨质疏松性骨折愈合过程中cbfa1和OCN的表达变化[J].中国骨质疏松杂志,2013,19(6):561-567.
[16] 赵可伟,潘君贻,肖林,等.中药骨康含药血清培养脂肪干细胞移植促进骨质疏松性骨折愈合的研究[J].热带医学杂志,2013,13(2):183-187,229.
[17] 宋瑄,林楠,李伟.鲑鱼降钙素鼻喷剂治疗女性围绝经期骨质疏松症临床疗效及安全性研究[J].中华保健医学杂志,2017,19(6):532-533.
[18] 王斌,罗毅文,胡年宏.骨康方含药血清诱导大鼠骨髓基质细胞向成骨细胞方向分化的实验研究[J].中国中医骨伤科杂志,2007,15(11):32-36.
[19] 余文珍,施红.复方中药含药血清研究现状[J].中医药通报,2003,2(2):113-116.
[20] 邵敏,赵静,王斌.不同中药含药血清对体外培养成骨细胞影响的实验研究[J].中国中医骨伤科杂志,2006,14(5):19-21.
[21] 邵敏,庄洪,赵静,等.含药血清对成骨细胞矿化结节及Ⅰ型胶原形成的影响[J].中医正骨,2003,15(4):3-4.
[22] 熊学华,余克强,刘庆思.中药骨康对去势大鼠骨吸收与骨形成影响的实验研究[J].现代康复,2001,5(11B):111-112.
[23] 李颖,黄宏兴,庄洪,等.中药骨康对去势大鼠骨质疏松作用机制的实验研究[J].中华中医药杂志,2009,24(2):160-163.
[24] 熊学华,冯新送,刘庆思.中药骨康对去势大鼠血清降钙素及骨生物力学影响的实验研究[J].中国中医骨伤科杂志,2005,13(3):7-9.
[25] Gu C, Fu L, Yuan X, et al. Promoting effect of pinostrobin on the proliferation, differentiation, and mineralization of murine pre-osteoblastic MC3T3-E1 cells[J].Molecules, 2017,22(10):1735.
[26] 万雷,黄宏兴,刘庆思,等.骨康对绝经后骨质疏松症患者骨密度、血清E2、SOD、MDA的影响[J].医药前沿,2012,2(14):93-94.