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浙江大学学报(农业与生命科学版)  2015, Vol. 41 Issue (1): 7-14    DOI: 10.3785/j.issn.1008-9209.2014.03.231
于张颖, 谌迪, 王一然, 沈立荣*
Effect of major royal jelly proteins (MRJPs) on  proliferation activity of Chang's  liver cell line and their mechanism of action
Yu Zhangying, Chen Di, Wang Yiran, Shen Lirong*
(School of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China)
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摘要: 将王浆主蛋白(major royal jelly proteins,MRJPs)添加到细胞培养基中,用MTT法比较纯MRJPs与胎牛血清(fetal bovine serum,FBS)、不同MRJPs/FBS比例对张氏肝细胞增殖的影响,用流式细胞仪检测MRJPs各处理对细胞周期的影响。结果表明:仅添加MRJPs不能促进细胞增殖;完全培养基(10% FBS)中MRJPs的最佳添加量为0.5 mg/mL;MRJPs(5 mg/mL)/FBS的添加比例为60/40时混合培养基对细胞的促增殖效果与完全培养基相比差异无统计学意义(P>0.05)。流式细胞仪检测结果显示,MRJPs与FBS配合使用可促使细胞S期及G0 /G1期比例增大;推测MRJPs的作用可能与促进DNA合成、前期RNA和核糖体合成有关。
Abstract: Major royal jelly proteins (MRJPs) are the soluble proteins in royal jelly, which account for 82%90% of total royal jelly proteins. MRJPs are consisted of ten members, i.e. MRJP1-MRJP9 and MRJPψ, whose biological functions include enhancing cell proliferation, inducing differentiation, modulating immune responses, accelerating wound healing, etc. MRJP1 with 57 ku in molecular mass was found possessing proliferation-promoting activities, enhancing proliferation of primary cultured rat hepatocytes and increasing albumin production in the absence of serum. MRJPs could significantly stimulate the proliferation and growth of endothelial progenitor cells (EPC). The recombinant MRJP1 could significantly stimulate growth of Tn-5B-4 cell from Trichoplusia ni, and affect cell shape and adhesion to the substrate. Similar activities were also found in normal human neonatal skin fibroblasts NB1 RGB cell, MC3T3-E1 cell and Jurkat cell, etc. All these findings showed the potential of MRJPs in cell culture as cell growth factor. Besides, it seemed that MRJP1 could promote liver regeneration and might have a cytoprotective action on hepatocytes. Here, we used Chang’s liver cell from human as object to observe the proliferation action of MRJPs on the cell line, and to explore the potential of MRJPs substituting fetal bovine serum (FBS) as growth factor; the active mechanism of MRJPs was also investigated. MRJPs were extracted by centrifuging from fresh royal jelly.The optimum concentration was selected by MTT assaying of the proliferation rates of the cell (PRC) cultured with serum-free mediums containing 0.05, 0.1, 0.2, 0.3 and 0.5 mg/mL of MRJPs at 5th day. The possibility of MRJPs with optimum concentration (0.5 mg/mL ) to replace FBS was investigated via comparison of the effects on PRC at the 2nd day, 5th day and 8th day in both serum-free and serum-existing mediums with a positive control containing 10% FBS and a negative control containing 10% phosphate buffer solution (PBS). It was showed that MRJPs could only be used to culture the cell line when it was mixed with FBS. Then, the effects of different MRJPs/FBS (M/F) ratios (0/100, 30/70, 60/40, 90/10, 100/0) on PRC at the 2nd day, 5th day and 8th day were investigated, respectively. It was shown that the M/F ratios with 30/70 and 60/40 were better for PRC of the cell line in relative to the complete medium with 10% FBS (M/F=0/100). The cell cycle distributions of the cell line cultured with different M/F ratios (0/100, 30/70, 60/40, 100/0) were detected by flow cytometry. It was shown that the proliferation index (PI), S phase and G0/G1 phase of the cell in the medium with M/F of 60/40 at the 5th day were not significantly different from the cell in complete medium (P>0.05), indicating that MRJPs might promote DNA synthesizing in S phase or RNA and protein synthesizing in G0/G1 phase. In conclusion, MRJPs mixed with FBS are suggested partially to replace FBS to culture Chang’s liver cell line in practice.
出版日期: 2015-01-20
CLC:  Q 253  
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于张颖, 谌迪, 王一然, 沈立荣. 王浆主蛋白(MRJPs)对张氏肝细胞的促增殖作用及其机制[J]. 浙江大学学报(农业与生命科学版), 2015, 41(1): 7-14.

Yu Zhangying, Chen Di, Wang Yiran, Shen Lirong. Effect of major royal jelly proteins (MRJPs) on  proliferation activity of Chang's  liver cell line and their mechanism of action. Journal of Zhejiang University (Agriculture and Life Sciences), 2015, 41(1): 7-14.


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