[1]马笃军 彭力平△ 王立新 余阗 裴军宇 李明华 徐宁达 肖伟.牛膝醇提物诱导兔骨髓间充质干细胞软骨定向分化的实验研究[J].中国中医骨伤科杂志,2017,25(02):6-11.
　MA Dujun PENG Liping WANG Lixin YU Tian PEI Junyu LI Minghua XU Ningda XIAO Wei.Experimental Research on Chondrogenic Differentiation of Bone Marrow Mesenchymal Stem Cells of Rabbits Induced by Alcohol Extractive of Achyranthis Bidentatae[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2017,25(02):6-11.
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牛膝醇提物诱导兔骨髓间充质干细胞软骨定向分化的实验研究()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第25卷
期数:
2017年02期
页码:
6-11
栏目:
实验研究
出版日期:
2017-02-15

文章信息/Info

Title:
Experimental Research on Chondrogenic Differentiation of Bone Marrow Mesenchymal Stem Cells of Rabbits Induced by Alcohol Extractive of Achyranthis Bidentatae
文章编号:
1005-0205(2017)02-0006-06
作者:
马笃军1 彭力平1△ 王立新1 余阗1 裴军宇1 李明华2 徐宁达3 肖伟1
1广州中医药大学深圳临床医学院(广东 深圳,518033) 2深圳北京大学-香港科技大学医学中心生物医学研究所 3广州中医药大学深圳医院 △通信作者 E-mail:plp001@sina.com
Author(s):
MA Dujun1 PENG Liping1△ WANG Lixin1 YU Tian1 PEI Junyu1 LI Minghua2 XU Ningda3 XIAO Wei1
1Shenzhen Clinical College,Affiliated to Guangzhou University of Traditional Chinese Medicine,Shenzhen 518033,Guangdong China; 2Biomedical Research Institute,Medical Center,Shenzhen Peking University-The Hong Kong University of Science and Technology,Shenzhen 518000,Guangdong China; 3Shenzhen Hospital of Guangzhou University of Traditional Chinese Medicine,Shenzhen 518034,Guangdong China.
关键词:
牛膝醇提物;  怀牛膝;  骨髓间充质干细胞;  软骨分化
Keywords:
alcohol extractive of achyranthis bidentatae;  achyranthis bidentatae;  bone marrow mesenchymal stem cells;  chondrogenic differentiation
分类号:
R-33
文献标志码:
A
摘要:
目的:观察牛膝醇提物诱导兔骨髓间充质干细胞(BMSCs)软骨定分化的作用。方法:密度梯度离心联合骨髓贴壁法分离培养新西兰大白兔BMSCs,取P3代细胞随机分成5组(空白组、完全诱导组、牛膝醇提物低剂量组、牛膝醇提物中剂量组、牛膝醇提物高剂量组),连续诱导培养21 d,倒置相差显微镜观察细胞形态,免疫细胞学检测Ⅱ型胶原蛋白荧光表达,qRT-PCR检测软骨分化标记基因Sox9、蛋白聚糖、Ⅱ型胶原mRNA表达情况,Western Blot检测Sox9、蛋白聚糖、Ⅱ型胶原蛋白表达水平。结果:牛膝醇提物高、中、低剂量组与空白组相比Ⅱ型胶原蛋白荧光阳性表达明显; 牛膝醇提物高、中剂量组与空白组相比软骨分化标记基因Sox9、蛋白聚糖、Ⅱ型胶原明显增加(P<0.05),Western Blot验证Sox9、蛋白聚糖、Ⅱ型胶原蛋白表达明显(P<0.05),其中牛膝醇提物高剂量组效果最佳(P<0.01),与完全诱导组比较差异无统计学意义(P>0.05)。结论:牛膝醇提物能够诱导兔BMSCs 成软骨分化,其具体作用机制有待进一步研究。
Abstract:
Objective:To observe the effect of chondrogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)of rabbits in vitro induced by alcohol extractive of Achyranthis bidentatae(AEAB).Methods:The BMSCs from New Zealand rabbits were isolated and cultivated by percoll density gradient centrifugation combined with bone marrow adherent.BMSCs at passage 3 were randomized divided into 5 groups:blank group,the complete induction group,low-dose treatment group by AEAB,middle-dose group therapy by AEAB,high-dose treatment group by AEAB.They were induced for 21 days.The morphology was observed by inverted microscope.The expression of type Ⅱ collagen fluorescence was detected by immune cytology; the expression of cartilage differentiation marker genes of Sox9,aggrecan,and type Ⅱ collagen mRNA were detected by QRT-PCR; the expression levels of Sox9,aggrecan,and type Ⅱ collagen were also tested by Western blot.Results:Compared with the blank group,the type Ⅱ collagen fluorescence was positive expressed in the treatment group(by AEAB treating of low dose,medium dose,and high dose).Compared with the blank group,the expression of the cartilage differentiation marker genes,such as Sox9,aggrecan,and type Ⅱ collagen mRNA in the treatment group were increased significantly(P<0.05).The Western blotting confirmed expression of the Sox9,aggrecan,and type Ⅱ collagen(P<0.05),best effect was in the high-dose treatment group by AEAB(P<0.01),and there was no significant difference compared with the complete induction group(P>0.05).Conclusion:AEAB can induce BMSCs of rabbit into chondrogenic differentiation in vitro,but its specific mechanism needs further studied.

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备注/Memo

备注/Memo:
基金项目:深圳市科技计划(卫生系统)项目课题(201302124) 深圳市福田区卫生公益性科研项目(FTWS201318)
更新日期/Last Update: 2017-09-08