川续断皂苷Ⅵ通过JNK信号通路促进骨髓间充质干细胞成骨分化  

Asperosaponin Ⅵ Promoting Differentiation of Bone Marrow Mesenchymal Stem Cells into Osteoblasts Through JNK Signaling Pathway

作　　者：黄媛[1] 徐艳[2] 易学良[1] 王雪 胡刚 唐雨蝶 金昊 张晓[2] HUANG Yuan;XU Yan;YI Xue-Liang;WANG Xue;HU Gang;TANG Yu-Die;JIN Hao;ZHANG Xiao(Chengdu Medical College,Chengdu 610500 Sichuan,China;Basic Medical Experiment Teaching Center of Chengdu Medical College,Chengdu 610500 Sichuan,China)

机构地区：[1]成都医学院,四川成都610500 [2]成都医学院基础医学实验教学中心,四川成都610500

出　　处：《广州中医药大学学报》2018年第5期887-893,共7页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：成都医学院2016大学生创新实验训练计划项目(编号:201613705023)

摘　　要：【目的】探讨川续断皂苷Ⅵ诱导大鼠骨髓间充质干细胞(BMSCs)向成骨细胞方向分化的机制。【方法】全骨髓贴壁法培养大鼠BMSCs。实验分为诱导组,1μmol/L川续断皂苷Ⅵ组,1μmol/L川续断皂苷Ⅵ+5μmol/L SP抑制组,1μmol/L川续断皂苷Ⅵ+10μmol/L SP抑制组,1μmol/L川续断皂苷Ⅵ+20μmol/L SP抑制组等5组。采用茜素红染色和骨钙素定量法检测细胞成骨分化程度;采用碱性磷酸酶(ALP)染色测定ALP活性;采用生物信息学方法预测c-Jun氨基末端激酶(JNK)相关因子。【结果】与诱导组比较,1μmol/L续断皂苷Ⅵ组茜素红染色明显增加,ALP活性增强,骨钙素表达量增加(P<0.05)。与1μmol/L川续断皂苷Ⅵ组比较,1μmol/L川续断皂苷Ⅵ+5μmol/L SP抑制组、1μmol/L川续断皂苷Ⅵ+10μmol/L SP抑制组、1μmol/L川续断皂苷Ⅵ+20μmol/L SP抑制组茜素红染色程度降低,ALP活性减弱,骨钙素含量显著减少(P<0.05)。生物信息学表明c-Jun和转录激活因子4(ATF4)、骨形态发生蛋白2(BMP2)之间存在直接的相互联系。【结论】川续断皂苷Ⅵ诱导BMSCs成骨分化的机制可能与JNK信号通路的激活有关。Objective To explore the mechanism for differentiation of osteoblasts induced by Asperosaponin Ⅵ in bone marrow mesenchymal stem cells（BMSCs）of rats. Methods BMSCs were cultured with whole bone marrow adherent method. The achieved cells were divided into 5 groups, namely induction group, 1 μmol/L asperosaponin Ⅵ group,1 μmol/L asperosaponin Ⅵ plus 5 μmol/L SP inhibition group,1 μmol/L asperosaponin Ⅵ plus 10 μmol/L SP inhibition group,1 μmol/L asperosaponin Ⅵ plus 20 μmol/L SP inhibition group. Alizarin red staining and osteocalcin quantitation were used to assay the degree of osteogenic differentiation. Alkaline phosphatase（ALP）staining was used to detect ALP activity. Bioinformatics methodology was used to predict the related factors of C-Jun N-terminal kinase（JNK）. Results Compared with the induction group, in 1 μmol/L asperosaponin Ⅵ group,the degree of alizarin red staining was deepen,the activity of ALP was increased,and the expression level of osteocalcin was significantly increased（P〈0.05）. Compared with 1 μmol/L asperosaponin Ⅵ group, the degree of alizarin red staining was shallower, the activity of ALP and the expression level of osteocalcin were decreased significantly in the other 3 asperosaponin Ⅵ plus SP inhibition groups（P〈0.05）.Bioinformatics indicated that there was a direct correlation among c-Jun,activating transcription factor 4（ATF4）and bone morphogenetic protein 2（BMP2）. Conclusion The mechanism for osteogenic differentiation of BMSCs induced by asperosaponin Ⅵ may be related to the activation of JNK signaling pathway.

关 键 词：骨质疏松症 川续断 川续断皂苷Ⅵ JNK信号通路 骨髓间充质干细胞 成骨分化 

分 类 号：R285.5[医药卫生—中药学]