流体切应力促永生化大鼠BRL-3A肝细胞增殖的研究  

Effect of fluid shear stress on proliferation of immortalized rats BRL-3A liver cell lines

作　　者：马心逸 喻智勇[1] 韦嘉[1] 马铎[1] 邓 刘巧云[1] 高则海[1] 蔡强[1] 支春平[1] 

机构地区：[1]昆明医科大学第四附属医院普通外科三科650021

出　　处：《中华实验外科杂志》2016年第6期1534-1537,共4页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金面上项目（81060041）;云南省科技厅一昆明医科大学联合基金（2014FB073）

摘　　要：目的观察不同大小流体切应力对永生化大鼠BRL-3A肝细胞株增殖动力学的影响。方法构建流体切应力加载细胞培养装置，以永生化大鼠BRL-3A肝细胞株为研究对象，实验分为未添加肝细胞生长因子（HGF）的A组和添加HGF的B组，各组加载0、12、24dyn／cm^2大小的流体切应力，采用直接细胞计数和分光光度计测定细胞计数试剂盒（CCK-8）的方法检测12、24、48、168h细胞增殖动力学的变化。结果未加载流体切应力时，B组细胞增殖优于A组；加载流体切应力越大，A组和B组细胞均增殖更明显；在同一时间点加载同样大小切应力时，B组细胞增殖由于A组（P〈0．05）。加载大小为24dyn／cm^2的切应力培养细胞24h，A和B组的细胞计数和CCK-8分光光度计吸光度（A）值均达到最高值，细胞计数分别为A组（10．73±2．54）×10^6个和B组（15．93±2．00）×10^6个（P〈0．05），CCK-8分光光度计A值分别为A组（2．04±0．51）和B组（3．09±0．40，P〈0．05）；24—72h逐渐下降，但仍高于未加载应力组（P〈0．05），168h则接近或略高于未加载应力组。结论流体切应力对永生化大鼠BRL-3A肝细胞株增殖有促进作用，而且，理化因素（流体切应力联合HGF）共同作用下肝细胞增殖更明显。Objective To investgate the effect of shear stress on proliferation of immortalized rat BRL-3A liver cell lines. Methods Cell culture device which could load with fluid shear stress was built, in which the immortalized rat BRL- 3A liver cell lines were cultured. The experiment was divided into group A （added hepatocyte growth factor （HGF）） and group B （did not add HGF） , in each grouP〈0, 12 dyn/cm2 and 24 dyn/cm2 fluid shear stress was loaded separately. Then the cell proliferation dynamics was detected by direct cell counting or cell counting kit-8 （CCK-8） technology in 12, 24, 48, 168 h. Results Without loading fluid shear stress, the proliferation of group B was better than group A ; the high- er the fluid shear stress was loaded, the more ceils proliferated; When loaded the same scale fluid shear stress at the same time point, the proliferation of group B was better than group A （ P 〈 0. 05 ）. With load- ing fluid shear stress of 24 dyn/cm2 at the 24 h time point, all groups reached the peak of cell proliferation, in which group B was still better than Group A（cell counting was （ 15.93 ±2.00） ×10^6 cells in group B and （ 10.73 ±2. 54）×10^6 cells in group A, as well as A of CCK - 8 was 3.09 ± 0.40 in group B and 2. 04 ±0. 51 in group A respectively （P 〈0. 05） ; The cell proliferation dropped gradually during 24 -72 h, but it still higher than the group which without loading fluid shear stress （ P 〈 0. 05 ）, it got close to or a lit- tle higher than the non -stress group at 168 h time point. Conclusion Fluid shear stress could promote the proliferation of immortalized rats BRL -3A liver cell lines, the effect of cell proliferation was more ap- parent under the interaction of physical and chemical factors （fluid shear stress combined with HGF）.

关 键 词：肝再生 永生化大鼠BRL-3A肝细胞株 流体切应力 肝细胞生长因子 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]