|
| 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 |
| |
| DOI：10.3969/j.issn.1006.7108.2014.07.001 |
| 中文关键词: 牛骨胶原肽 成骨细胞 分化 |
| 英文关键词:BCP Osteoblasts Differentiation |
| 基金项目: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 |
|
| 摘要点击次数: 959 |
| 全文下载次数: 754 |
| 中文摘要: |
| 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对喊性磷酸酶(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。 |
| 英文摘要: |
| 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 cells (0.193 ±0. 021. P < 0. 01) at the 14th day, indicating that BCP could promote the mineralization in HOB cells. Conclusion BCP plays a positive role in osteoblast differentiation and the mineralized bone matrix formation. Taking all the experiments together, our study indicates a molecular mechanism for the potential prevention and treatment of osteoarthritis and osteoporosis. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |