CAJ | 학술논문

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.
목적:탐토갈근소대대서성골세포체외증식적영향.방법:신생SD대서,분리로개골성골세포진행원대、전대배양;취생장상태량호적제3대세포진행실험;도치상차현미경관찰세포형태특점급생장상태;I형효원면역세포화학염색、감성린산매염색、개결절염색등방법진행성골세포감정;이분별함유1×10-5mol/mL지1×10-10mol/mL농도적갈근소배양기작위실험조;불함갈근소적배양기작위대조조,관찰접충제1、3、5、7천4개시간점성골세포생장상태.통과CCK-8법검측성골세포적생장、증식정황,병회제생장곡선.결과:CCK-8법검측성골세포종제3천기세포증식수량개시증가,재제5、7천시,증식도1×10-10mol/L지1×10-7mol/L범위내,차수착갈근소농도적증고,성골세포증식작용증강병정제량의뢰성.결론:갈근소가능통과자격소수체개도촉진성골세포증식급골형성작용.당갈근소농도재1×10-10mol/L지1×10-7mol/L농도범위내시,기대성골세포촉증식작용최강.
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. Immunohistochemistry was used to examine the expression of typeI 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 1í10-5 mol/mL to 1í10 -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 cells grew fast obviously. The proliferation of cells ranging from 1í10-10 mol/L to 1í10-7 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 1í10-10 mol/L to 1í10-7 mol/L Puerarin was the most accelerated significantly.