临床剂量浓度透明质酸对骨髓间充质干细胞旁分泌及过表达VEGF165作用影响的实验研究  

Effect of paracrine and over- expression VEGF165 of bone marrow mesenchymal stem cell under the intervention of sodium hyaluronate culture

作　　者：张聪[1] 刘洪美[2] 李庆伟[1] 陈国武[1] 梁啸[1] 孟纯阳[1] 

机构地区：[1]济宁医学院附属医院脊柱外科,山东济宁272000 [2]济宁医学院附属医院病理教研室,山东济宁272000

出　　处：《中国矫形外科杂志》2014年第9期839-842,共4页Orthopedic Journal of China

基　　金：山东省自然科学基金资助项目(编号:ZR2010HM090)

摘　　要：[目的]观察临床剂量浓度透明质酸干预骨髓间充质干细胞（bone maltow mesenchymal stem cells，BMSCs）后对其生长、旁分泌和过表达VEGFl65蛋白水平的影响。[方法]将细胞生长曲线测定与细胞分泌VEGFl65水平测定均分为4组：A组（DMEM／F12），B组（HA），C组（HA＋Ad—VEGF165），D组（Ad—VEGF165），每组6个复孔。采用密度梯度离心法分离BMSCs和流式细胞术并进行细胞表型鉴定。取第3代BMSCs做为实验对象，Ad—VEGF165转染BMSCs后和或与HA复合培养6h，然后观察细胞形态、生长曲线、VEGFl65分泌水平变化。[结果]各组细胞形态未见明显差异和变化；C组与D组细胞生长曲线OD值在第1～5d均高于A组与B组；A组（34．524-2．13）与B组（34．90±1．78）均有较低水平的旁分泌VEGF65蛋白，而Ad—VEGF165转然后的C组（1487．29±36．08）与D组（1474．48±49．13）VEGF165分泌水平明显增高，但两组差异无统计学意义，而与A组、B组相比较差异均有统计学意义。[结论]治疗剂量透明质酸钠对BMSCs生长、增殖、旁分泌及过表达VEGF165蛋白无明显影响。[ Objective] To determine the effect of therapeutic level of hyaluronic acid on cultured BMSCs for its growing status and levels of secreting VEGFI65 protein. [ Methods] The cell growth curve assay and cell secretion VEGF165 level measurement were divided into four groups, i.e. , group A （ DMEM/F12 ）, group B （ HA ）, group C （ HA ＋ Ad - VEGF165）, group D （Ad - VEGF165） . BMSCs was separated by density gradient centrifugation and cell phenotype was iden- tified by flow cytometry, respectively. The third passage BMSCs as subject, the cell morphology, growth curve and VEGF165 secretion levels changes were observed while BMSCs transfected by Ad - VEGF165 and/or in HA cultured after 6 h. [ Results] All groups of cells morphology were not obviously changed and OD value in group C and group D in 1 - 5 days were higher than in group A and group B. Group C （ 1 487. 29 ± 36.08） and group D （ 1 474.48 ± 49. 13 ） VEGF165 secretion level significantly increased, there was no statistically significant difference in these groups but statistical difference was found as they were compared with group A and group B. [ Conclusion] The effects of therapeutic dose HA on cultured BMSCs growing status, prolif- eration and VEGF165 secretion are minimal.

关 键 词：透明质酸 骨髓间充质干细胞 血管内皮生长因子165 旁分泌蛋白 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]