摘要
目的考察骨肽原液对胎儿骨髓间充质干细胞(fetal marrow mesenchymal stem cells,F-BMSCs)成骨分化的影响,并对其相关作用机制进行初步探讨。方法选取4个不同厂家生产的骨肽原液,对F-BMSCs干预的同时进行成骨诱导。诱导后在不同时间点进行ALP活性及茜素红染色检测,使用定量PCR进行骨分化标志物基因OPN、OCN、Runx2、ALP表达的检测,使用Western blot进行特异性标记蛋白Runx2和OCN表达的检测。结果经成骨诱导后,与诱导组相比,骨肽供试品组厂家C骨肽原液处理的F-BMSCs ALP活性明显增加(P<0.05);厂家B、L骨肽原液处理的F-BMSCs ALP活性高于诱导组,但差异无显著性(P>0.05)。F-BMSCs经茜素红染色,肉眼观察厂家B、C、L骨肽原液处理的F-BMSCs红色明显增强;显微镜下厂家B、C、L骨肽原液处理的F-BMSCs茜素红染色阳性结节状覆盖面积明显多于诱导组。厂家B骨肽原液处理的F-BMSCs成骨分化标志物基因OPN、OCN、Runx2、ALP的mRNA表达量增加,明显高于诱导组;Runx2、OCN蛋白表达亦明显高于诱导组。结论骨肽原液对F-BMSCs的成骨分化具有明显促进作用。
Aim To investigate the effect of ossotide stock solution on osteogenic differentiation of fetal bone marrow mesenchymal stem cells,and to explore its related mechanism.Methods Osteoinduction was performed while treating with F-BMSCs by ossotide from four different manufacturers.ALP activity and alizarin red staining were performed.Quantitative expressions of OPN,OCN,Runx2 and ALP were detected by quantitative PCR.The expression of specific marker proteins Runx2 and OCN was assessed by Western blot.Results After osteogenic induction,compared with induction group,ALP activity of manufacturer C inossotide test group significantly increased(P<0.05);ALP activity of manufacturers B,L was higher than that of induction group,but there was no significant difference(P>0.05).The F-BMSCs were stained with alizarin red,the color of the manufacturers B,C and L in test group were significantly enhanced.Under the microscope,the alizarin red staining positive nodular coverage area of manufacturers B,C and L significantly increased.The mRNA expression levels of osteogenic differentiation marker genes OPN,OCN,Runx2,and ALP increased in ossotide test group(manufacturer B),and were significantly higher than those in induction group;Runx2 and OCN protein expressions were also significantly higher than those in induction group.Conclusion The ossotide stock solution has a marked promoting effect on the osteogenic differentiation of F-BMSCs.
作者
张媛
吴彦霖
纳涛
杨泽岸
胡文言
高华
ZHANG Yuan;WU Yan-lin;NA Tao;YANG Ze-an;HU Wen-yan;GAO Hua(Division of Pharmacology,Institute for Chemical Drug Control,National Institutes for Food and Drug Control,Beijing 102629,China;China Biochemical Pharmaceutical Industry Association,Beijing 100068,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2020年第6期791-797,共7页
Chinese Pharmacological Bulletin
基金
国家药典委员会国家药品标准提高研究课题(No 2018H001)。
关键词
骨肽原液
间充质干细胞
成骨分化
碱性磷酸酶
茜素红
骨钙素
ossotide stock solution
mesenchymal stem cells
osteogenic differentiation
alkaline phosphatase
alizarin red
osteocalcin
