虾肽对MC3T3-E1成骨细胞增殖、分化及矿化的影响  

Effects of Shrimp Peptide on the Proliferation, Differentiation and Mineralization of MC3T3-E1 Osteoblasts

作　　者：相兴伟 邵婉 舒聪涵 李锦弘 陈慧[1] 邓尚贵 周宇芳 Xiang Xingwei;Shao Wan;Shu Conghan;Li Jinhong;Chen Hui;Deng Shanggui;Zhou Yufang(College of Food Science and Technology,Zhejiang University of Technology,Key Laboratory of Marine Fishery Resources Exploitment&Utilization of Zhejiang Province,Hangzhou 310014;Zhejiang Marine Development Research Institute,Zhoushan 316021,Zhejiang;College of Food and Pharmacy,Zhejiang Ocean University,Zhoushan 316022,Zhejiang)

机构地区：[1]浙江工业大学食品科学与工程学院,浙江省深蓝渔业资源高效开发利用重点实验室,杭州310014 [2]浙江省海洋开发研究院,浙江舟山316021 [3]浙江海洋大学食品与药学学院,浙江舟山316022

出　　处：《中国食品学报》2022年第10期116-125,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家重点研发计划项目(2020YFD0900901);浙江省基础公益研究计划项目(LGN20C200002)。

摘　　要：目的:从红虾副产物中提取虾肽,研究其促MC3T3-E1成骨细胞增殖、分化及矿化的活性。方法:用MTT法检测不同浓度虾肽对MC3T3-E1成骨细胞存活率的影响;诱导成骨细胞分化,在3 d和7 d时,测定其碱性磷酸酶(ALP)活性,在7 d和14 d时,测定其细胞骨钙素(OCN)及Ⅰ型胶原蛋白(COL-Ⅰ)含量,21 d时茜素红染色测定细胞诱导培养矿化程度;采用qPCR和Western blot方法检测虾肽对OPG/RANKL/RANK信号通路中骨形成关键基因和蛋白OPG、RANKL以及RUNX2表达的影响。结果:虾肽质量浓度为0.02,0.05,0.1 mg/mL时,显著促进MC3T3-E1细胞的增殖;ALP、OCN及COL-Ⅰ的含量相较于对照组均增加,矿化结节面积显著增大(P<0.05);此外,虾肽上调了ALP、OCN、COL-Ⅰ基因的表达,促进OPG和RUNX2基因及蛋白表达的水平,抑制RANKL基因及蛋白表达的水平。结论:虾肽能促进MC3T3-E1成骨细胞增殖分化与矿化,激活OPG/RANKL/RANK信号通路,从而促进成骨细胞的分化及骨形成。Objective: To extract shrimp peptides from the by-products of red shrimp, and study its activity in promoting the proliferation, differentiation and mineralization of MC3T3-E1 cells. Methods: Firstly, the MTT method was used to detect the effect of different concentrations of shrimp peptides on the survival rate of MC3T3-E1 cells. After the concentration was selected, the osteoblasts were induced to differentiate and culture, and the alkaline phosphatase(ALP) activity was measured at 3 and 7 days of culture. Measure the content of osteocalcin(OCN) and type Ⅰ collagen(COL-Ⅰ) at 7 d and 14 d. Alizarin red staining is used to detect the degree of mineralization of cells induced and cultured at 21 d.QPCR and Western blot are used to study the expression of key genes and the proteins OPG, RANKL and RUNX2 in bone formation in the OPG/RANKL/RANK signaling pathway. Results: Shrimp peptide mass concentrations of 0.02, 0.05 mg/mL, and 0.1 mg/mL significantly promoted cell proliferation, ALP activity, OCN and COL-Ⅰ content increased compared to the control group, and the area of mineralized nodules increased significantly(P < 0.05). Shrimp peptide upregulates the expression of ALP, OCN, and COL-Ⅰ genes, promotes the expression of OPG and RUNX2 genes and protein levels, and inhibits the expression of RANKL genes and protein levels. Conclusion: Shrimp peptide can promote the proliferation, differentiation and mineralization of MC3T3-E1 osteoblasts, and activate the OPG/RANKL/RANK signaling pathway to promote osteoblast differentiation and bone formation.

关 键 词：虾肽 MC3T3-E1成骨细胞 分化 矿化 

分 类 号：TS254.9[轻工技术与工程—水产品加工及贮藏工程]