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[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 marrow mesenchymal stem cells,BMSCs)后对其生长、旁分泌和过表达VEGF165蛋白水平的影响。[方法]将细胞生长曲线测定与细胞分泌VEGF165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,AdVEGF165转染BMSCs后和或与HA复合培养6 h,然后观察细胞形态、生长曲线、VEGF165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1~5 d均高于A组与B组;A组(34.52±2.13)与B组(34.90±1.78)均有较低水平的旁分泌VEGF165蛋白,而Ad-VEGF165转然后的C组(1 487.29±36.08)与D组(1 474.48±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 observe the effect of hyaluronic acid on the growth, paracrine and overexpression of VEGF165 protein in bone marrow mesenchymal stem cells (BMSCs) treated with clinical dose of hyaluronic acid. [Methods] The cell growth curve and the level of VEGF165 secreted by cells were divided into 4 groups: group A (DMEM / F12), group B (HA), group C (HA + Ad- VEGF165), group D ), Each set of 6 complex hole. BMSCs were isolated by density gradient centrifugation and flow cytometry was performed and cell phenotypes were identified. The third generation BMSCs were taken as experimental subjects. AdVEGF165 was transfected into BMSCs or cultured with HA for 6 h. Then the changes of cell morphology, growth curve and VEGF165 secretion were observed. [Results] There were no significant differences and changes in cell morphology between groups. The OD values of cell growth curve in groups C and D were higher than those in groups A and B on the 1st to 5th day, while those on the 1st and 5th groups in group A (34.52 ± 2.13) and group B 34.90 ± 1.78) had lower levels of paracrine VEGF165 protein, while the secretion of VEGF165 in group C (1 487.29 ± 36.08) and group D (1 474.48 ± 49.13) after Ad-VEGF165 switch was significantly higher, but there was no significant difference between the two groups Statistical significance, but with A group, B group compared with the difference was statistically significant. [Conclusion] Sodium hyaluronate at the therapeutic dose has no significant effect on the growth, proliferation, paracrine and overexpression of VEGF165 protein in BMSCs.