摘要
研究了从鸡蛋黄中提取的黄高磷蛋白(PV)(0,50,100,200μg/mL和500μg/mL)对RAW264.7细胞增殖、周期、凋亡的影响,结果发现低浓度PV对RAW264.7细胞有促进增殖和抑制凋亡的作用,当PV质量浓度100μg/mL时效果最好。采用CCK-8法表征增殖活力,结果发现PV质量浓度为100μg/mL时增殖活力高出对照组64%~76%。分析PV作用24 h和48 h后对细胞周期的影响,不同浓度的PV对细胞增殖均有促进作用,与对照组相比低质量浓度PV(50,100、200μg/mL)处理细胞后S期细胞的百分比含量、增殖指数PI值大。凋亡结果表明,50,100,200μg/mL PV组凋亡率显著低于空白对照组,500μg/mL时凋亡率高于空白组。另外,在抗酒石酸酸性磷酸酶(TRAP)染色细胞分化形态学及TRAP活性方面,不同浓度的PV均可显著减少破骨细胞数量,降低TRAP活性(P<0.05),且呈浓度依赖性。其中,200μg/mL为最佳抑制质量浓度。PV并非通过抑制RAW264.7细胞生长而是通过显著降低TRAP的活性来抑制RAW264.7细胞分化成为破骨细胞。
The effects of different concentrations of PV separated from egg yolk(0,50,100,200 and 500μg/mL)on the proliferation,cell cycle and cell apoptosis of RAW264.7 cells for different time were studied.It was found that low concentration of PV could promote proliferation and inhibit the apoptosis of RAW264.7 cells.The effect of 100μg/mL PV concentration which can promote the proliferation and inhibit apoptosis is best.CCK-8 method was used to characterize the proliferation of RAW264.7 cells.It was found that the proliferation of RAW264.7 cells showed two aspects.The cell proliferation was promoted at low concentration,the cell proliferation was inhibited at high concentration.The results showed that the cell viability was 64%-76%higher than that of the control group at 100μg/mL.By analyzing the effect of PV on the cell cycle,found that the percentage of PV in the S phase was significantly higher than that in the control group(P<0.05),100μg/mL PV has the best effect.The apoptotic effects of PV group at 50,100 and 200μg/mL were significantly lower than that of control group.When the concentration of PV was 500μg/mL,the apoptotic rate was higher than that of the control group,which indicated that low concentration of PV could inhibit the apoptosis and high concentration of PV could induce apoptosis.In addition,different concentrations of PV can significantly reduce the number of osteoclasts and the activity of TRAP(P<0.05),thus inhibiting the RAW264.7 cell differentiation to form osteoclasts,and in a concentration-dependent manner,the best inhibitory concentration was 200μg/mL.In conclusion,PV was not by inhibiting the growth of RAW264.7 cells,but by inhibiting the differentiation of RAW264.7 cells significantly reducing the activity of TRAP,thereby inhibiting the formation of osteoclasts.
作者
介怡琳
李晓云
纪胜男
赵前程
马美湖
黄茜
Jie Yilin;Li Xiaoyun;Ji Shengnan;Zhao Qiancheng;Ma Meihu;Huang Xi(College of Food Science and Technology,Huazhong Agricultural University,National Research and Development Centre for Egg Processing,Key Laboratory of Egg Processing,Ministry of Agriculture and Rural Affairs,Wuhan 430070)
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2020年第8期29-40,共12页
Journal of Chinese Institute Of Food Science and Technology
基金
“十三五”国家重点研发计划项目(2018YFD0400302)
国家自然科学基金项目(31471602)
国家蛋鸡产业技术体系岗位科学家项目(CARS-40-K24)。
