葛根素对高浓度糖皮质激素环境下MC3T3-E1前成骨细胞生物学性能的影响  

Effects of Puerarin on Biological Properties of MC3T3-E1 under High Concentration of Glucocorticoid Circumstance

作　　者：陈振宇 潘颖菁[2] 张艳[3] 贺玺[3] 黄达鸿[2] CHEN Zhenyu;PAN Yingjing;ZHANG Yan;HE Xi;HUANG Dahong(Chengnan Branch, Foshan Stomatology Hospital, School of Stomatology and Medicine, Foshan University, Foshan 528000,China;Center of Prosthodontics and Oral Implantology, Foshan Stomatology Hospital, School of Stomatology and Medicine, Foshan University, Foshan 528000, China;Hospital of Stomatology, Jilin University, Changchun 130021,China)

机构地区：[1]佛山科学技术学院附属口腔医院·佛山市口腔医院城南分院,广东佛山528000 [2]佛山科学技术学院附属口腔医院·佛山市口腔医院修复种植科,广东佛山528000 [3]吉林大学口腔医院,吉林长春130021

出　　处：《口腔医学研究》2020年第6期544-547,共4页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:51541206)。

摘　　要：目的:通过体外细胞培养以及相关干预,探讨葛根素对高浓度糖皮质激素环境下MC3T3-E1生物学性能的影响。方法:体外培养MC3T3-E1前成骨细胞,分为一对照组与五组实验组,在高浓度糖皮质激素(10^-6 mmol/L)环境下使用梯度浓度葛根素(0、1×10^-5、1×10^-6、1×10^-7、1×10^-8、1×10^-9 mol/L)进行干预;使用CCK-8试剂盒分别检测培养1 d、4 d、7 d时的细胞增殖、碱性磷酸酶试剂盒检测细胞培养1 d、4 d、7 d时的ALP活性,培养7 d时实时定量PCR检测细胞OPG mRNA的表达,培养24 h鬼笔环肽染色并在激光共聚焦显微镜下观察细胞骨架形态变化。结果:CCK-8结果显示,对照组与各实验组差异具有统计学意义(P<0.05),培养4 d时各实验组之间比较1×10^-7mol/L组细胞增殖最明显(P<0.05);ALP检测结果显示,同一时间点各实验组ALP表达均有提升,且与对照组差异有统计学意义(P<0.05);激光共聚焦鬼笔环肽染色结果显示,在1×10^-7mol/L组细胞铺展面积最大,结构清晰;PCR结果显示,各实验组OPG mRNA的表达量均高于对照组并与对照组差异有统计学意义(P<0.05)。结论:葛根素能够在高浓度糖皮质激素环境下促进MC3T3-E1增殖、伸展及分化。Objective:To investigate the effect of puerarin on the biological properties of MC3T3-E1 under high concentration of glucocorticoid circumstance through in vitro cell culture and related interventions.Methods:MC3T3-E1 preosteoblasts were cultured in vitro and divide into control group and five experimental groups,which were intervened with gradient concentration of puerarin(0,10^-5,10^-6,10-7,10^-8,10^-9 mol/L)under high concentration of glucocorticoid circumstance(10-6 mmol/L).Cell proliferation was detected after 1 d,4 d,and 7 d of cell culture by using the CCK-8 kit,activity of ALP was detected after 1 d,4 d,and 7 d of cell culture by using alkaline phosphatase kit,and the expression of OPG mRNA was detected by quantitative real-time PCR after 7 d of cell culturing.The cells were stained with phalloidin after 24 h of culture and observed the cytoskeletal morphology changes under the confocal laser microscope.Results:CCK-8 showed that the difference between the control group and the experimental group was statistically significant(P<0.05),and the most obvious effect of cell proliferation was 10^-7 mol/L group on the 4th day(P<0.05).ALP activity test showed that the expression of ALP in all the experimental groups increased at the same time and there was statistical significance in the difference between the experimental group and the control group(P<0.05).The results of confocal laser scanning indicated that,in group of 10^-7 mol/L,the cells spread more widely and the structure was clear.PCR results showed that the expression of OPG mRNA in each experimental group was higher than that in the control group(P<0.05).Conclusion:Puerarin,under high concentration of glucocorticoid circumstance,can promote cell proliferation,extension,and differentiation of MC3T3-E1 cells.

关 键 词：葛根素 糖皮质激素 成骨细胞 增殖 分化 

分 类 号：R28[医药卫生—中药学]