牛骨胶原肽对成骨细胞分化的影响  

Effect of bovine collagen peptides on the differentiation of osteoblasts

作　　者：刘俊丽[1] 张兵[2] 宋淑军[1] 马铭[2] 司少艳[1] 王毅虎[2] 吴继功[1] 郭燕川[2] 

机构地区：[1]解放军第306医院特种病医学实验中心,北京100101 [2]中国科学院理化技术研究所,光化学转换与功能材料重点实验室,北京100190

出　　处：《中国骨质疏松杂志》2014年第7期723-727,共5页Chinese Journal of Osteoporosis

基　　金：中国科学院理化技术研究所协作课题(Grant No.LHS-DBSW2013-01);总装后勤部课题资助项目

摘　　要：目的研究3 mg/ml牛骨胶原肽(Bovine collagen peptides,BCP)对人成骨细胞(Human osteoblast,HOB),小鼠前成骨细胞系MC3T3分化的影响。方法 Western blot检测MC3T3细胞中BCP对Runx2表达的影响;分离培养HOB,利用对硝基苯磷酸比色法检测3 mg/ml BCP对碱性磷酸酶(Alkaline phosphatase,ALP)含量的影响;骨钙素(osteocalcin,OC)ELISA法测定BCP对OC含量的影响;茜素红矿化染色检测BCP对HOB矿化,然后用5%高氯酸进行脱色,用吸光度法检测BCP对人成骨细胞矿化程度。结果 Western blot检测表明,14 d后BCP处理的MC3T3细胞中Runx2蛋白表达水平(0.178±0.201)与CN组(0.146±0.582,P>0.05)比较,有增高趋势。ALP染色结果表明,BCP混合物处理组的ALP的染色面积(33859±8221)在第10d与CN组(19900±2796)相比,显著增加(P<0.05),表明BCP混合物能促进人成骨细胞早期的ALP表达量。BCP混合物处理组的OC吸光度值(0.137±0.014)在第14d高于CN组(0.086±0.023,P<0.05),表明BCP混合物能促进人成骨细胞晚期阶段的OC含量。茜素红矿化染色结果表明,3mg/ml BCP能显著促进HOB的矿化骨基质的形成。当使用5%的高氯酸脱色后,BCP处理组在490 nm处测吸光度值(0.579±0.093)显著高于CN组(0.193±0.021,P<0.01),表明BCP混合物能促进人成骨细胞的矿化。结论 BCP在成骨细胞分化和矿化骨基质的形成中发挥了积极作用。综合上述结果,本文为BCP混合物在骨关节炎和骨质疏松症潜在的预防和治疗提供了分子机理。Objective To investigate the effect of 3 mg/ml bovine collagen peptides （BCP） on the differentiation of human osteoblasts （HOB） and mouse MC3T3-E1 pre-osteoblasts. Methods The effect of BCP on the expression of runt-related transcription factor 2 （Runx2） in MC3T3-E1 cells was detected using Western blotting. HOBs were isolated and cultured with BCP. The concentration of alkaline phosphatase （ALP） was detected using colorimetric p-nitrophenyl phosphate assay. The concentration of osteocalcin （OC） was detected using ELISA. After Alizarin red staining and decolorization with 5% perchloric acid, the effect of BCP on the mineralization in HOBs was detected using spectrophotometric method. Results The result of Western blotting revealed that the expression of Runx2 in BCP-treated MC3T3 cells at the 14th day was higher than that in control cells （0. 178 ± 0. 201 vs. 0. 146 ± 0. 582）. There was an increasing trend, but no significance was observed （P 〉 0.05 ）. The results of ALP staining showed that the ALP activity in the BCP-treated cells （33859 ± 8221.7） was much higher than that in the control cells （19900 ±2796, P 〈0. 05） at the 10th day, indicating that BCP could promote the ALP activity in MC3T3-E1 cells at the early stage, The OC activity in BCP-treated cells （0. 137 ±0. 014） was much higher than that in the control cells （0. 086 ±0. 023, P 〈 0.05） at the 14th day, indicating that BCP could promote the OC activity in MC3T3-E1 cells at late stage. Calcium deposit test showed that BCP significantly increased mineralization in HOB cells at the 14th day. After decolorization with 5% perchloric acid, the absorbance at 490 nm in BCP-treated cells （0. 579 ± 0. 093 ） was much higher than that in the control ceils （0. 193 ± 0. 021, P 〈 0.01 ） at the 14th day, indicating that BCP could promote the mineralization in HOB ceils. Conclusion BCP plays a positive role in osteoblast differentiation and the mineralized bone matrix formation. Taking all the experim

关 键 词：牛骨胶原肽 成骨细胞 分化 

分 类 号：R855.22[医药卫生—航空、航天与航海医学]