梅花鹿鹿茸I型胶原诱导骨髓基质干细胞成骨分化及其分子机制的研究  

An experimental study on the procedure and mechanism of sika pilose antler type I collagen induces BMSCs differentiation into osteoblasts

作　　者：王艳双[1,2] 罗速[2] 张大方[1] 曲晓波[1] 李娜[1] 谭寅凤 李枫[2] 王秀丽[2] 

机构地区：[1]长春中医药大学,吉林长春130117 [2]北华大学,吉林吉林132013

出　　处：《时珍国医国药》2013年第12期3056-3059,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81073158)

摘　　要：目的 探讨梅花鹿鹿茸Ⅰ型胶原诱导骨髓基质干细胞向成骨细胞分化及其分子机制.方法 采用CCK-8法选择诱导分化和促成骨的最佳浓度;采用分光光度法检测ALP活性,并用改良Gomori法进行ALP染色,采用茜素红法进行钙化结节染色.采用RT-PCR法检测成骨分化的关键因子RunX2 mRNA的表达,成骨标志物ALP mRNA的表达,并进行半定量分析.结果 2.5g·L-1的鹿茸Ⅰ型胶原促进BMSCs的增殖使ALP活性降低,5～40 g·L-1的鹿茸Ⅰ型胶原抑制BMSCs的增殖,呈时间、剂量依赖性,5g·L-1的鹿茸Ⅰ型胶原使ALP活性增加显著且促进钙化结节的形成.RT-PCR检测5g·L-1的鹿茸Ⅰ型胶原上调RunX2、ALP基因的表达,而2.5g·L-1的鹿茸Ⅰ型胶原则下调RunX2、ALP基因的表达.结论 梅花鹿鹿茸Ⅰ型胶原可以通过上调RunX2基因的表达诱导BMSCs向OB分化,且与浓度密切相关.Objective To investigate the procedure and mechanism by which velvet antler type Ⅰ collagen induces BMSCs differ- entiation into osteoblasts. Methods The best induced differentiation and osteogenic effects concentration were measured by CCK - 8 method ; alkaline phosphatase activity was detected by spectrophotometry and alkaline phosphatase staining with improved Go- mori method, calcified nodules staining with Allizarin red. The expression of Runx2 mRNA of the key factor osteogenic differentia- tion was measured by RT - PCR, the expression of ALP mRNA of osteoblast marker was measured by RT - PCR, and semi - quantitative analysis. Results The type Ⅰ collagen （ 2.5 g·L^ -l ） promoted proliferation of BMSCs, decreased alkaline phospha- tase activity; type Ⅰ collagen （ 5 - 40 g ·L^ -l ） restrained proliferation of BMSCs, and presented time - dependent and dose - de- pendent;type Ⅰ collagen （5 g ·L^ -l ） significantly promoted alkaline phosphatase activity and increased form of calcified nodules. The results demonstrated that type Ⅰcollagen （2.5 g·L^ -l） increased the expression of Runx2 and ALP , but type Ⅰ collagen （2. 5 g ·L^ -l ） reduced the expression of Runx2 and ALP. Conclusion The velvet antler type Ⅰ collagen induced BMSCs differentia- tion into osteoblasts by increased the expression of Runx2, and was bound up with concentration.

关 键 词：梅花鹿鹿茸I型胶原 BMSCs CCK-8 RUNX2 ALP 

分 类 号：R285[医药卫生—中药学]