畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (12): 2745-2752.doi: 10.11843/j.issn.0366-6964.2018.12.024

• 临床兽医 • 上一篇    下一篇

羊骨髓肽及其钙螯合物对成骨细胞增殖和活性的影响

金淑秀1, 王钰慧2, 霍乃蕊1*, 郑明学1, 韩克光1, 古少鹏1, 田文霞1, 张鼎1   

  1. 1. 山西农业大学 动物科技学院, 太谷 030801;
    2. 山西农业大学 食品科学与工程学院, 太谷 030801
  • 收稿日期:2018-06-11 出版日期:2018-12-23 发布日期:2018-12-23
  • 通讯作者: 霍乃蕊,女,山西平遥人,博士,教授,主要从事动物疾病预防与动物性食品安全研究,E-mail:tgnrhuo@163.com
  • 作者简介:金淑秀(1994-),女,山西祁县人,硕士生,主要从事兽医公共卫生与人畜共患传染病防治的研究,E-mail:517509072@qq.com
  • 基金资助:

    国家自然科学基金(31201347);山西省回国留学人员项目(2014-042)

Function of Bone Marrow Peptide and Its Calcium-Chelated Form in the Proliferation and the Osteogenic Activity of Cultured Osteoblasts

JIN Shu-xiu1, WANG Yu-hui2, HUO Nai-rui1*, ZHENG Ming-xue1, HAN Ke-guang1, GU Shao-peng1, TIAN Wen-xia1, ZHANG Ding1   

  1. 1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;
    2. College of Food Science and Engineering, Shanxi Agricultural University, Taigu 030801, China
  • Received:2018-06-11 Online:2018-12-23 Published:2018-12-23

摘要:

成骨细胞在骨代谢过程中发挥成骨作用,笔者制备羊骨髓肽钙螯合物(MP-Ca),探究其对体外培养成骨细胞的增殖及活性的影响,并将其效果同羊骨髓肽(MP)进行比较。制备的MP-Ca经傅里叶红外光谱仪进行扫描鉴定;从新生SD大鼠颅骨中分离纯化的成骨细胞经碱性磷酸酶(ALP)和茜素红染色鉴定;在培养液中添加不同浓度的受试物,CCK-8法确定最佳作用浓度。以17-β雌二醇(102 μg·mL-1)为阳性对照,通过测定各组细胞OD450 nm和ALP、骨钙素(BGP)的含量,考察MP、MP-Ca在最适作用浓度(103μg·mL-1)下对成骨细胞增殖和活性的影响。结果表明,MP和MP-Ca的红外光谱图未完全重叠,吸收峰发生了明显的位移变化,说明MP与CaCl2反应形成了MP-Ca。分离的成骨细胞经ALP染色呈阳性,胞内可见大量黑褐色颗粒;茜素红染色后细胞呈深红色,并形成钙化结节。MP-Ca处理组的OD450 nm和ALP、BGP水平均显著(P<0.05或P<0.01)高于MP处理组,但均显著(P<0.05或P<0.01)低于雌激素组。综上,尽管效果不及雌激素,MP-Ca和MP均可促进体外培养成骨细胞的增殖并增强其成骨活性,效果MP-Ca优于MP。

Abstract:

Osteoblasts play bone-forming roles in the process of bone metabolism. Calcium-chelated bone marrow peptides (MP-Ca) was prepared and its effects on the proliferation and osteogenic activity of osteobalst (OB) were investigated and compared with calcium-free peptides of sheep bone marrow (MP). Prepared MP-Ca was identified via Fourier transform infrared spectrometer. The osteoblasts isolated from the skull of newborn SD rats were identified through alkaline phosphatase (ALP) and alizarin red staining. Different concentrations of test substance, including the positive control 17 β-estradiol, were added to the medium and the CCK-8 method was used to determine the optimal concentration for the subsequent study. The proliferation and the osteogenic activity of OB were expressed as OD450 nm and the content of ALP and osteocalcin (BGP) in cell lysate prepared from MP-Ca, MP, estrogen treated OB cells at optimal concentrations. MP and MP-Ca showed no completely overlapped infrared spectra and apparent shifts of absorption peaks were observed, which indicated that MP-Ca was different form MP and it was prepared successfully. After ALP staining the isolated and purified OB cells appeared positive reaction. The results showed that OD450 nm, ALP and BGP levels of the MP-Ca treated cells were significantly higher than those of the MP treatment (P<0.05 or P<0.01), but they were significantly lower than those of 17β-estradiol treated group (P<0.05 or P<0.01). It can be concluded that although inferior to 17β-estradiol, MP and MP-Ca were effective in promoting the proliferation of osteoblasts in vitro and enhance their bone formation activity, and MP-Ca was superior to MP.

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