透骨消痛颗粒水提与醇提物含药血清干预骨髓基质干细胞活性的实验研究  

Tougu Xiaotong Granule water/alcohol extract-containing serum interferes the activity of bone marrow stromal stem cells

作　　者：刘伯龄[1] 李西海[2] 梁珪清[1] 陈齐勇[1] 吴广文[2] 李照辉[1] 

机构地区：[1]福建中医药大学附属福州中西医结合医院(福州市第二医院),福建省福州市350007 [2]福建中医药大学中西医结合研究院,福建省福州市350108

出　　处：《中国组织工程研究》2012年第41期7717-7722,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助(81001530);福建省自然科学基金(2011J05074);福州市科技计划项目(2011-S-67-1)~~

摘　　要：背景:骨髓基质干细胞根据环境而有不同的分化路径,在特定的环境中有分化为透明软骨细胞的趋势,可能为治疗骨性关节炎提供新思路。目的:观察透骨消痛颗粒水提与醇提物含药血清对骨髓基质干细胞活性的影响。方法:制备透骨消痛颗粒水、醇提物,108只SD大鼠随机摸球法均分为低、中、高剂量对照组、水、醇提物组,腹主动脉采血制备含药血清。取4周龄SD大鼠四肢骨髓基质干细胞,第3代同步化后用含药血清进行干预,MTT检测其活性,流式细胞仪检测其周期分布,Real time PCR检测Cyclin D1 mRNA表达,Western Blot检测Cyclin D1蛋白表达。结果与结论:透骨消痛颗粒水、醇提物含药血清干预48h后,软骨细胞的A值中高剂量组明显高于对照组(P<0.01,P<0.05);各组G0/G1期细胞,水、醇提物组明显低于对照组(P<0.01),水提物组明显低于醇提物组(P<0.05);各组增殖指数及Cyclin D1 mRNA及蛋白表达水、醇提物组明显高于对照组(P<0.01),水提物组明显高于醇提物组(P<0.05)。说明透骨消痛颗粒水醇提物含药血清通过上调Cyclin D1的表达,加速骨髓基质干细胞周期的进程,从而促进其增殖,且透骨消痛颗粒水提物的效果优于醇提物。BACKGROUND：Bone marrow stromal stem cells have multilineage differentiation capacity and can differentiate into transparent chondrocytes,which can provide new thoughts for treatment of osteoarthritis.OBJECTIVE：To investigate the effects of Tougu Xiaotong Granule water/alcohol extract-containing serum on the activities of bone marrow stromal stem cells.METHODS：Tougu Xiaotong Granule water/alcohol extracts were prepared.A total of 108 SD rats were randomly divided into control group,Tougu Xiaotong Granule water extract group and Tougu Xiaotong Granule alcohol extract group.Then each group was divided into three subgroups as per low,medium,and high dose.Abdominal aorta blood was taken to prepare drug-containing serum.Passage 3 bone marrow stromal stem cells from 4-week-old SD rats were interfered with Tougu Xiaotong Granule water/alcohol extract-containing serum,and then cell activity was detected by MTT.Cell cycle was determined by flow cytometry.Cyclin D1 mRNA expression was detected by real-time PCR.Cyclin D1 protein expression was determined by western blot analysis.RESULTS AND CONCLUSION：After 48 hours of intervention by Tougu Xiaotong Granule water/alcohol extract,the ratio of cells in the G0/G1 period in the Tougu Xiaotong Granule water extract group and Tougu Xiaotong Granule alcohol extract group was significantly lower than that in the control group（P 0.01）,the ratio of cells in the G0/G1 period in the Tougu Xiaotong Granule water extract group was significantly lower than that in the Tougu Xiaotong Granule alcohol extract group（P 0.05）.Proliferation index and cyclin D1 mRNA and protein expression in the Tougu Xiaotong Granule water extract group were significantly higher than in the control group（P 0.01）,and these indices were significantly higher in the Tougu Xiaotong Granule water extract group than in the Tougu Xiaotong Granule alcohol extract group（P 0.05）.These findings suggest that Tougu Xiaotong Granule water/alcohol extract-containing serum promotes the proliferation of bon

关 键 词：透骨消痛颗粒 骨性关节炎 含药血清 骨髓基质干细胞 细胞周期 干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]