骨康胶囊组方药物对成骨细胞SaOS-2增殖分化及矿化的影响
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摘要
目的:研究骨康胶囊组方药物续断、三七、芭蕉根对体外培养人成骨细胞Sa OS-2增殖、分化和矿化的影响。方法:以不含药培养基培养Sa OS-2作为对照组,给药组Sa OS-2分别用1、10、100、200、400、800 mg/L续断、芭蕉根及三七水提物作用,MTT法测定Sa OS-2细胞增殖情况,磷酸苯二钠比色法检测碱性磷酸酶(ALP)活性,ELISA法检测钙离子和骨钙素(OTC)的分泌以及用茜素红染色法测定成骨细胞矿化结节数。结果:与对照组比较,续断各浓度水提物可显著促进Sa OS-2细胞的增殖、增强ALP活性且对细胞Ca2+的分泌有促进作用(P<0. 05或P <0. 01),浓度为100、200 mg/L的续断水提物对Sa OS-2细胞分泌OTC能力及对细胞的矿化有促进作用(P <0. 05或P <0. 01);三七水提物除了浓度为200 mg/L时有明显促进OTC分泌的作用(P <0. 05)外,浓度为10、100 mg/L的三七水提物及各浓度的芭蕉根水提物对成骨细胞增殖、分化和矿化作用均不明显(P>0. 05)。结论:骨康胶囊治疗骨质疏松作用可能与其组方续断的促进成骨细胞的增殖、分化及矿化的作用有关。
Objective: To investigate in vitro effects of components( including Dipsacus asper,Panax notoginseng,Musa basjoo Sieb) of Gu Kang capsule on cell proliferation,differentiation and mineralization of human osteoblast SaOS-2 cells. Methods: SaOS-2 cells were cultured and treated with different concentrations of water extracts of Dipsacus asper,Panax notoginseng,Musa basjoo Sieb. Cell proliferation was determined using MTT assay. Alkaline phosphatase( ALP) activity was measured by benzene disodium phosphate colorimetriy,and secretions of Ca2 +,osteocalcin( OTC) were detected by ELISA assay. Osteoblast mineralization nodules were measured by alizarin red staining. Results: Compared with untreated group,water extract of Dipsacus asper at 10,100 and 200 mg/L concerntrations significantly promoted the proliferation of osteoblasts,enhanced ALP activity and Ca2 +secretion in vitro( P< 0. 05,P < 0. 01). Water extract of Dipsacus asper at 100 and 200 mg/L concerntrations could promote the secretion of OTC and osteoblasts mineralization( P < 0. 05,P < 0. 01),while its effects were not obvious at 10 mg/L concentration( P > 0. 05). Water extract of Panax notoginseng at 200 mg/L concerntration could significantly promote OTC secretion( P < 0. 05). However,water extract of Panax notoginseng at 10 and 100 mg/L showed no significant effect on the proliferation,differentiation and mineralization of SaOS-2 cells( P > 0. 05). Furthermore,water extract of Musa basjoo Sieb had no significant effect on proliferation,differentiation and mineralization of SaOS-2 cells. Conclusions: The anti-osteoporosis effect of Gukang Capsule may be associated with its roles of Dipsacus asper in promoting the proliferation,differentiation and mineralization of osteoblasts SaOS-2.
Objective: To investigate in vitro effects of components( including Dipsacus asper,Panax notoginseng,Musa basjoo Sieb) of Gu Kang capsule on cell proliferation,differentiation and mineralization of human osteoblast SaOS-2 cells. Methods: SaOS-2 cells were cultured and treated with different concentrations of water extracts of Dipsacus asper,Panax notoginseng,Musa basjoo Sieb. Cell proliferation was determined using MTT assay. Alkaline phosphatase( ALP) activity was measured by benzene disodium phosphate colorimetriy,and secretions of Ca2 +,osteocalcin( OTC) were detected by ELISA assay. Osteoblast mineralization nodules were measured by alizarin red staining. Results: Compared with untreated group,water extract of Dipsacus asper at 10,100 and 200 mg/L concerntrations significantly promoted the proliferation of osteoblasts,enhanced ALP activity and Ca2 +secretion in vitro( P< 0. 05,P < 0. 01). Water extract of Dipsacus asper at 100 and 200 mg/L concerntrations could promote the secretion of OTC and osteoblasts mineralization( P < 0. 05,P < 0. 01),while its effects were not obvious at 10 mg/L concentration( P > 0. 05). Water extract of Panax notoginseng at 200 mg/L concerntration could significantly promote OTC secretion( P < 0. 05). However,water extract of Panax notoginseng at 10 and 100 mg/L showed no significant effect on the proliferation,differentiation and mineralization of SaOS-2 cells( P > 0. 05). Furthermore,water extract of Musa basjoo Sieb had no significant effect on proliferation,differentiation and mineralization of SaOS-2 cells. Conclusions: The anti-osteoporosis effect of Gukang Capsule may be associated with its roles of Dipsacus asper in promoting the proliferation,differentiation and mineralization of osteoblasts SaOS-2.
引文
[1]郭丽艳,王金兰,谭音玲,等.沙格列汀对高糖环境下成骨细胞增殖、分化和凋亡的影响[J].山东医药,2017,57(28):16-19.
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[8]HOWLETT C R,CAVE J,WILLIAMSON M,et al. Min-eralization in in vitro cultures of rabbit marrow stromalcells.[J]. Clin Orthop Relat Res,1986,213(213):251-263.
[9]户小伟,李巍,韦达隆,等.碱性成纤维细胞生长因子对兔骨髓间充质干细胞向成骨细胞分化的影响[J].山东医药,2016,56(48):8-11.
[10]唐琪,陈莉丽,严杰.骨碎补提取物促小鼠成骨细胞株MC3T3-E1细胞增殖、分化和钙化作用的研究[J].中国中药杂志,2004,29(2):164-168.
[11]程志安,吴燕峰,黄智清,等.续断对成骨细胞增殖、分化、凋亡和细胞周期的影响[J].中医正骨,2004,16(12):1-3.
[12]吴丽萍,陶天遵,石义刚,等.三七总甙对成骨细胞增殖、分化及OPG表达影响的研究[J].中国骨质疏松杂志,2004,10(2):239-241.
[13]MCCORMICK R K. Osteoporosis:integrating biomarkersand other diagnostic correlates into the management ofbone fragility.[J]. Alternative Medicine Review,2007,12(2):113-145.
[2]翟远坤,牛银波,潘亚磊,等.柚皮苷对体外培养乳鼠颅骨成骨细胞增殖和分化成熟的影响[J].中国中药杂志,2013,38(1):105-111.
[3]易显富,张泽月,程蕊,等. 1α,25-二羟维生素D3对体外培养的成骨细胞分化及矿化成熟影响[J].浙江临床医学,2014,16(2):169-172.
[4]李娜,颜冬梅,张金莲,等.炮制方法对补骨脂中抗骨质疏松效应成分含量的影响[J].世界科学技术-中医药现代化,2017,19(1):127-132.
[5]安亚兰,王建舫,许水明,等.补骨脂水提液及补骨脂素对体外培养成骨细胞的影响[J].畜牧兽医学报,2009,40(2):266-271.
[6]刘晓艳,董莉,刘亭,等.酢浆草提取物对成骨细胞增殖及分化的影响[J].中国实验方剂学杂志,2015,21(1):117-120.
[7]杨健,陆定艳,彭潇,等.不同来源、不同炮制方法的补骨脂对成骨细胞Sa OS-2骨形成功能的影响[J].海南医学,2018,29(15):2140-2143.
[8]HOWLETT C R,CAVE J,WILLIAMSON M,et al. Min-eralization in in vitro cultures of rabbit marrow stromalcells.[J]. Clin Orthop Relat Res,1986,213(213):251-263.
[9]户小伟,李巍,韦达隆,等.碱性成纤维细胞生长因子对兔骨髓间充质干细胞向成骨细胞分化的影响[J].山东医药,2016,56(48):8-11.
[10]唐琪,陈莉丽,严杰.骨碎补提取物促小鼠成骨细胞株MC3T3-E1细胞增殖、分化和钙化作用的研究[J].中国中药杂志,2004,29(2):164-168.
[11]程志安,吴燕峰,黄智清,等.续断对成骨细胞增殖、分化、凋亡和细胞周期的影响[J].中医正骨,2004,16(12):1-3.
[12]吴丽萍,陶天遵,石义刚,等.三七总甙对成骨细胞增殖、分化及OPG表达影响的研究[J].中国骨质疏松杂志,2004,10(2):239-241.
[13]MCCORMICK R K. Osteoporosis:integrating biomarkersand other diagnostic correlates into the management ofbone fragility.[J]. Alternative Medicine Review,2007,12(2):113-145.