小鼠脊髓背根神经节细胞与人微血管内皮细胞共培养对细胞增殖的影响  

Effect on cell proliferation of mouse dorsal root ganglion cells co-culture with human microvascular endothelial cells

作　　者：原泉[1] 程扬[2] 杨亚帆[1] 孙立[3] 郭金明[1] 

机构地区：[1]中国医科大学附属盛京医院骨科,沈阳110004 [2]沈阳市骨科医院 [3]中国医科大学附属第一医院肾内科

出　　处：《中华实验外科杂志》2015年第1期135-138,共4页Chinese Journal of Experimental Surgery

基　　金：辽宁省自然科学基金资助项目（201102252）;辽宁省社会发展攻关计划资助项目（201225094）;辽宁省高等学校优秀人才支持计划资助项目（LNET）

摘　　要：目的 观察小鼠脊髓背根神经节细胞（DRGC）与人微血管内皮细胞（HMVEC）共培养对细胞增殖的影响,并探讨其机制.方法 分离培养小鼠DRGC,并与HMVEC进行共培养,设为DRGC组、HMVEC组、DRGC＋ HMVEC组.噻唑蓝（MTT）法检测细胞增殖,实时定量聚合酶链反应（Real-time PCR）及Western blot检测血管内皮生长因子（VEGF）、神经生长因子（NGF）、增殖细胞核抗原（PCNA）和细胞周期蛋白D1（Cyclin Dl） mRNA及蛋白的表达.结果 细胞培养24h后,DRGC＋ HMVEC组相对于DRGC组和HMVEC组的细胞增殖率分别为136.29％和137.45％,细胞增殖能力显著提高（P＜0.05）.VEGF、NGF、PCNA和Cyclin Dl mRNA和蛋白的表达在DRGC＋HMVEC组中最高,与其他两组比较差异有统计学意义（P＜0.05）.结论 共培养DRGC和HMVEC能够促进VEGF和NGF的表达,并可能通过调控PCNA和Cyclin D1的表达促进共培养体系细胞增殖能力.Objective Mouse dorsal root ganglion cells （DRGC） co-culture with human microvascular endothelial cells （HMVEC）,then study the effect on cell proliferation and investigate the possible mechanisms.Methods Separate mouse DRGC,then co-culture with HMVEC.Experiment was set as DRGC,HMVEC,DRGC ＋ HMVEC.Cell proliferation was detected by methyl thiazol tetrazolium （MTF）approach.Expression levels of vascular endothelial growth factor （VEGF）,nerve growth factor （NGF）,proliferating cell nuclear antigen （PCNA） and Cyclin D1 mRNA and protein were measured by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting.Results DRGC was separated successfully.After 24 hours co-culture of DRGC with HMVEC,cell relative proliferation rate was 136.29% compared with DRGC single culture,and 137.45% compared with HMVEC single culture.Cell proliferation ability of DRGC ＋ HMVEC group increased significantly （P 〈 0.05）,and mRNA and protein expression level of VEGF,NGF,PCNA and Cyclin D1 are also significantly increased （P 〈0.05）.Conclusion VEGF and NGF promote proliferation of mouse DRGC and HMVEC in co-culture by regulating the expression of PCNA and Cyclin D1 possibly.

关 键 词：小鼠脊髓背根神经节细胞 人微血管内皮细胞 神经生长因子 血管内皮细胞生长因子 增殖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]