摘要
本研究旨在探讨异槲皮苷对犬骨髓间充质干细胞(cBMSCs)成骨分化的影响及其机制。研究分为对照组和异槲皮苷组,采用MTT法检测细胞活力;茜素红染色检测cBMSCs的钙质沉积情况;荧光定量PCR检测成骨相关基因碱性磷酸酶(ALP)和特异AT序列结合蛋白2(SATB2)的表达;ELISA法检测细胞上清液中ALP含量;荧光定量PCR检测JNK信号通路相关基因SOS1(son of sevenless homolog 1)和激活转录因子2(ATF 2)的表达。结果显示,与对照组相比,添加异槲皮苷可显著促进cBMSCs的增殖,使cBMSCs的矿化面积增大,ALP和SATB2的mRNA水平及细胞上清液中ALP蛋白含量升高,SOS1和ATF2 mRNA水平降低。本研究结果表明,异槲皮苷对cBMSCs的增殖及成骨分化有明显促进作用,且促进cBMSCs成骨分化的机制可能是通过抑制JNK信号通路实现的。
This study aimed to explore the effect of isoquercitrin on the osteogenic differentiation of canine bone marrow mesenchymal stem cells(cBMSCs)and its mechanism.The experiment was divided into the control group and the isoquercitrin group.Cell viability was detected by MTT method.Alizarin red staining was used to detect calcium deposits in cBMSCs.Fluorescence quantitative PCR was used to detect osteogenic-related genes alkaline phosphatase(ALP)and the expression of specific AT sequences binding protein 2(SATB2).ELISA method was used to detect ALP content in cell supernatant.The expression of JNK signaling pathway related genes SOS1(son of sevenless homolog 1)and activating transcription factor 2(ATF2)was detected by fluorescence quantitative PCR.The results showed that compared with the control group,the addition of isoquercitrin could significantly promote the proliferation of cBMSCs,increase the mineralization area of cBMSCs,increase the mRNA levels of ALP and SATB2 and the ALP protein content in the cell supernatant,and decrease the mRNA levels of SOS1 and ATF2.The results of this study indicate that isoquercitrin can significantly promote the proliferation and osteogenic differentiation of cBMSCs,and the mechanism of promoting the osteogenic differentiation of cBMSCs may be achieved by inhibiting the JNK signaling pathway.
作者
汪娅媛
郭娟
田仰清
严涵
昂艳芬
闫晓霞
严玉霖
WANG Yayuan;GUO Juan;TIAN Yangqing;YAN Han;ANG Yanfen;YAN Xiaoxia;YAN Yulin(College of Animal Medical,Yunnan Agricultural University,Kunming 650201,China;Yunnan Xishuangbanna Prefecture Animal Disease Prevention and Control Center,Jinghong,Yunnan 666100,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2021年第11期2190-2195,共6页
Chinese Journal of Veterinary Science
基金
校企合作基金资助项目(2017530117000288)。
