摘要
目的:探讨葛根素对大鼠成骨细胞体外增殖的影响。方法:新生SD大鼠,分离颅盖骨成骨细胞进行原代、传代培养;取生长状态良好的第3代细胞进行实验;倒置相差显微镜观察细胞形态特点及生长状态;I型胶原免疫细胞化学染色、碱性磷酸酶染色、钙结节染色等方法进行成骨细胞鉴定;以分别含有1×10-5mol/mL至1×10-10mol/mL浓度的葛根素培养基作为实验组;不含葛根素的培养基作为对照组,观察接种第1、3、5、7天4个时间点成骨细胞生长状态。通过CCK-8法检测成骨细胞的生长、增殖情况,并绘制生长曲线。结果:CCK-8法检测成骨细胞从第3天起细胞增殖数量开始增加,在第5、7天时,增殖到1×10-10mol/L至1×10-7mol/L范围内,且随着葛根素浓度的增高,成骨细胞增殖作用增强并呈剂量依赖性。结论:葛根素可能通过雌激素受体介导促进成骨细胞增殖及骨形成作用。当葛根素浓度在1×10-10mol/L至1×10-7mol/L浓度范围内时,其对成骨细胞促增殖作用最强。
Objective： To explore the effect of Puerarin on the proliferation of rat osteoblast in vitro. Methods： Newborn SD rat OBs were separated and cultured in vitro with the calvaria. The OBs of the third generation were cultured. The morphological characteristics of OBs were observed by inverted phase contrast microscope, lmmunohistochemistry was used to examine the expression of type I collagen. Alkaline phosphatase（ALP） and calcifying nodule were stained. The osteoblasts cultured for 1, 3, 5 and 7 day were divided into experimental group at the culture medium doses ranging from 1×10^-5 mol/mL to 1 ×10^-10 mol/mL Puerarin and control group at the common culture medium without any Puerarin. The growth and proliferation ability was analyzed by CCK-8 method and growth curve was drawn. Results： For the OBs growth and proliferation ability by CCK-8 method, three days later, the ceils grew fast obviously. The proliferation of eells ranging from 1×10^-10 mol/L to 1×10^-7 mol/L Puerarin was accelerated significantly after cultured for 5 and 7 days. Conclusion： Puerarin promotes proliferation of OBs and bone formation by the meditation of estrogen receptor. The osteoblasts of OBs treated doses ranging from 1×10^-10 mol/L to 1×10^-7mol/L Puerarin was the most accelerated significantly.
出处
《中医药导报》
2013年第3期77-79,共3页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
关键词
葛根素
大鼠
成骨细胞
增殖
体外实验
Puerarin
Rat
Osteoblast
Proliferation
Vitro experiment
