摘要
目的研究钙、磷对体外培养成骨细胞(OB)增殖、分化及矿化的作用,探讨钙、磷对骨代谢的影响。方法在体外培养OB的基础上,分别添加1、2、4mmol/L钙、1、2、4mmol/L磷及2mmol/L钙+1mmol/L磷(钙磷比2∶1)、1mmol/L钙+2mmol/L磷(钙磷比1∶2)。检测细胞增殖、碱性磷酸酶(ALP)活性、骨桥蛋白(OPN)和Ⅰ型胶原(Col-Ⅰ)含量及其mRNA表达情况。结果与对照组比,饲料中添加4mmol/L钙从D2开始促进OB增殖,差异显著(P<0.05),添加1、2mmol/L钙从D5开始差异显著(P<0.05),而钙磷比2:1与1:2到D8差异才显著(P<0.05)。除1mmol/L钙组外,各试验组从D2起均抑制细胞内ALP活性(P<0.05),在D5抑制其mRNA表达(P<0.01),在D2、D8促进其mRNA表达(P<0.05)。除1mmol/L钙组外,各试验组在D2均促进Col-Ⅰ分泌(P<0.01),但钙各组和1mmol/L磷组在D5、D8及2mmol/L磷组和钙磷比(1:2)组在D8均抑制其分泌(P<0.05)。各试验组,在D2、D8均促进Col-ⅠmRNA表达(P<0.01),在D5除钙磷比(1:2)组外均抑制其mRNA表达(P<0.01)。各试验组促进OPNmRNA表达(P<0.01),除1、2mmol/L钙组外,各实验组从D2起均促进其分泌(P<0.0)。结论添加钙及2种钙磷比(2:1,1:2)均能抑制成骨细胞早期分化,诱导其增殖及重叠生长,促进其基质成熟及矿化,而磷对其增殖作用不明显。
Objective To clarify the influence of Ca and P on bone metabolism by investigating theeir effects on proliferation, differentiation and mineralization of osteoblasts (OBs) in vitro. Method OBs were cultured in media with Ca(1, 2,4 mmol/L),P(1, 2,4 mmol/L),Ca:P 1:2 (1 mmol/L Ca+2 mmol/L P), and Ca:P 2:1 (2 mmol/L Ca+1 mmol/L P). Cell proliferation, activity of alkaline phosphatase (ALP), content of osteopontin (OPN) and collagen type I (Col-I) of OBs, and their corresponding mRNA expression were measured. Results Compared with control group,Ca at 4 mmol/L promoted OBs proliferation significanty (P〈0.05) from D2,while Ca at 1 and 2 mmol/L from D5(P〈0.05),and Ca :P (1:2, 2:1) until D8. (P〈0.05). All groups except 1 mmol/L Ca inhibited the activity of ALP (P〈0.05) on D2, and inhibited its mRNA expression on D5 (P〈0.01), but enhanced its mRNA expression on D2 and D8 (P〈0.05). All groups except 1 mmol/L Ca promoted Col-I secretion (P〈0.01) on D2 but all Ca groups and 1 mmol / L P group on D5,D8, and 2 mmol/L P, Ca:P (1:2) group on D8 inhibited its secretion (P〈0.05). All groups promoted Col-I mRNA expression on D2、D8 (P〈0.01),but inhibited its mRNA expression on D5 (P〈0.01) except Ca:P (1:2) group. All groups enhanced OPN secretion (P〈0.05) except 1,2 mmol/L Ca group on D2. All groups promoted OPN mRNA expression (P〈0.01). Conclusion Adding Ca, and two Ca:P(2∶1,1∶2)can inhibit early differentiation and promote proliferation of OBs and induce matrix maturation, but P shows no significant effects on OBs proliferation.
出处
《营养学报》
CAS
CSCD
北大核心
2009年第6期537-542,共6页
Acta Nutrimenta Sinica
基金
国家自然科学基金(No30440050
30571364)
江苏省自然科学基金(NoBK2008214)
关键词
SD大鼠
成骨细胞
增殖
分化
钙
磷
SD rat osteoblast proliferation mineralization calcium phosphorus