黄芪多糖缓释纤维对成骨细胞MC3T3-E1增殖分化的影响  

Effects of Astragalus Polysaccharides Sustained-release Fibers on the Proliferation and Differentiation of Osteoblast Cells MC3T3-E1

作　　者：王相峰[1] 刘童斌[2] 陈亚丹[1] 付秀娟[1] 

机构地区：[1]吉林大学第二医院,长春130041 [2]吉林大学口腔医院,长春130041

出　　处：《中国药房》2015年第4期484-487,共4页China Pharmacy

摘　　要：目的:研究黄芪多糖(AP)缓释纤维对成骨细胞MC3T3-E1增殖分化的影响。方法:将AP分别与左旋聚乳酸(PLLA)结合制备成AP质量浓度分别为0、25、50、100μg/ml的静电纺丝缓释纤维,即PLLA、PLLA/AP1、PLLA/AP2、PLLA/AP3,以异硫氰酸荧光素为指示剂于荧光显微镜下观察MC3T3-E1细胞在上述4种缓释纤维上的黏附情况;采用紫外分光光度法在490 nm波长处测定PLLA/AP1、PLLA/AP2、PLLA/AP3中AP的含量,考察其21 d内的体外释放情况;采用MTT法检测4种缓释纤维作用24、48、72 h时MC3T3-E1细胞的增殖情况;采用酶标仪法检测4种缓释纤维对MC3T3-E1细胞作用3、5、7 d时的碱性磷酸酶(ALP)活性表达情况。结果:镜下观察PLLA纤维连续,较多空隙,表面光滑;PLLA/AP1、PLLA/AP2、PLLA/AP3纤维呈纵横交错的三维、多空网络结构,且3种AP载药量下纤维形态未见明显改变。PLLA/AP1、PLLA/AP2、PLLA/AP3能持续释放AP 21 d,且最初的24 h内释药呈突释状态。与PLLA比较,PLLA/AP2、PLLA/AP3能促进MC3T3-E1细胞的增殖,PLLA/AP1仅作用48 h时能促进MC3T3-E1细胞的增殖,差异具有统计学意义(P<0.05);PLLA/AP1、PLLA/AP2、PLLA/AP3均能增强MC3T3-E1细胞中ALP活性表达,差异具有统计学意义(P<0.05),且与AP浓度呈正相关。结论:AP缓释纤维具有一定的缓释作用,且对MC3T3-E1细胞的增殖、分化具有剂量依赖性的促进作用。OBJECTIVE： To study the effects of Astragalus polysaccharides （AP） sustained-release fibers on the proliferation and differentiation of osteoblast cells MC3T3-E1. METHODS： 0, 25, 50 and 100 μg/ml AP were dispersed into Poly-/-lactide acid （PLLA） to fabricate electrospun fibers, i.e. PLLA, PLLA/AP1, PLLA/AP2 and PLLA/AP3. The adhesion of MC3T3-E1 to 4 kinds of sustained-release fiber were observed by fluorescence microscope using fluorescein isothiocyanate as indicator. The contents of AP in PLLA/AP1, PLLA/AP2 and PLLA/AP3 were determined by UV spectrophotometry at 490 nm. The in vitro release of AP was investigated within 21 d. The proliferation of MC3T3-E1 treated with 4 kinds of sustained-release fibers for 24, 48 and 72 h were measured by MTT assay. The expression of alkaline phosphatase （ALP） of MC3T3-E1 cells was detected by ELISA after treat- ed with 4 kinds of sustained-release fibers for 3, 5 and 7 d. RESULTS： PLLA fibers were continuous and smooth in appearance, and had multiple gaps. PLLA/AP1, PLLA/AP2 and PLLA/AP3 fibers showed the maze of three-dimensional multiple-gap reticular structure, and there was no obvious change of fiber morphology under the drug-loading amount of 3 kinds of AE AP could be con- tinuously released by PLLA/AP1, PLLA/AP2 and PLLA/AP3 for 21 d, and showed burst release within first 24 h. Compared with PLLA, PLLA/AP2 and PLLA/AP3 could promote the proliferation of MC3T3-E1, and the promotion effect of PLLA/AP1 on the proliferation of MC3T3-E1 only needed 48 h; there was statistical significance （P〈0.05）. PLLA/AP1, PLLA/AP2 and PLLA/AP3 could enhance the expression of ALP in MC3T3-E1, which was positively associated with AP dose; there was statistical signifi- cance （P〈0.05）. CONCLUSIONS： AP can effectively control the release of AP and promote the proliferation and differentiation of MC3T3-E1 in dose-dependent manner.

关 键 词：黄芪多糖 左旋聚乳酸 缓释纤维 MC3T3-E1细胞 增殖 分化 

分 类 号：R944.9[医药卫生—药剂学] R965[医药卫生—药学]