摘要
目的研究麦角甾苷对大鼠骨髓基质细胞增殖和成骨分化的影响及机制。方法密度梯度离心法分离培养大鼠骨髓基质细胞并鉴定,细胞随机分为高、中、低浓度实验组和对照组,分别用100,50,25和0μmol·L^(-1)的麦角甾苷干预培养。培养24,4,72 h后,用细胞计数法(CCK-8)检测细胞增殖情况。培养24,48,96 h后进行碱性磷酸酶活性检测,72 h后进行碱性磷酸酶染色。以蛋白质印迹法(Western blot)及实时聚合酶链式反应(Real time PCR)分别检测成骨相关蛋白及基因表达情况。结果培养72 h后,高剂量实验组骨髓基质细胞增殖能力为1.22±0.05,低于对照组的1.50±0.09,差异有统计学意义(P<0.05)。培养24,48,96 h后,高、中、低浓度实验组碱性磷酸酶表达量较对照组增加,差异均有统计学意义(均P<0.05)。培养72 h后,碱性磷酸酶染色结果显示,高、中、低浓度实验组培养皿颜色均深于对照组。Western blot检测结果显示,高、中、低浓度实验组骨形态发生蛋白2(BMP2)、Smad1、Smad5、Smad8、Runx2、Osterix蛋白表达水平均高于对照组(P<0.05),而各剂量实验组间差异无统计学意义(P>0.05)。Real time PCR检测结果显示,高、中、低浓度实验组BMP2、Smad1、Smad5、Smad8、Runx2、Osterix基因表达均高于对照组(P<0.05),而各浓度实验组间差异无统计学意义(P>0.05)。结论麦角甾苷对体外培养的大鼠骨髓基质细胞增殖并无显著影响,并可以通过BMP2-Smads-Runx2-Osterix通路促进BMSCs成骨分化。
Objective To study the effects of verbascoside on the proliferation and osteogenic differentiation of rat bone marrow stromal cells(BMSCs） and its mechanism. Methods BMSCs were obtained and identified from rat bone marrow by density gradient centrifugation.BMSCs were randomly divided into high,medium,low dose test groups and control group,and treated with 100,50,25,0 μmol·L-1 verbascoside. The cell counting kit-8(CCK-8） method was used to evaluate the BMSCs proliferation at 24,48,72 h after administration the medication. The alkaline phosphatase(ALP） activities were detected at 24,48,96 h after medication. The ALP staining was performed after 72 h of medication. Western blot and real time-polymerase chain reaction(PCR） were used to detect the osteogenic related proteins and genes expression. Results After 72 h,the proliferation of BMSCs in high dosetest group was 1. 22 ± 0. 05,lower than that in control group,which was 1. 50 ± 0. 09(P〈0. 05）. The ALP expression levels in high,medium and low dose test groups were higher than control group at 24,48,96 h. After 72 h,ALP staining showed that the color of culture dishes in high,medium and low dose test groups were deeper than that in control group. Western blot showed that osteogenic related proteins of bone morphogenetic protein-2(BMP2）,Smad1,Smad5,Smad8,Runx2,Osterix in high,medium and low dose test groups were higher than those in control group(P〈0. 05）,but there was no significant difference among those three test groups(P〈0. 05）. Real time-PCR showed that the osteogenic related genes of BMP2,Smad1,Smad5,Smad8,Runx2,Osterix expression levels in high,medium and low dose test groups were higher than those in control group(P〈0. 05）. However,there was no significant difference among three test groups(P〉0. 05）. Conclusion The verbascoside at different concentrations has no obvious effect on BMSCs proliferation and promote osteogenic differentiation of BMSCs through BMP2-Smads-Runx2-Osterix pathway.
作者
宋欧荻
秦书俭
於绍龙
屈惠莹
田原
SONG Ou- di;QIN Shu- jian;YU Shao - long;Qu Hui - ying;TIAN Yuan(Preclinical Medicine College, Jinzhou Medical University, Jinzhou 121001, Liaoning Province, China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2018年第10期1206-1209,共4页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(31170930)
关键词
麦角甾苷
骨髓基质细胞
增殖
成骨分化
verbascoside
bone marrow stromal cells
proliferation
osteogenic differentiation
