CAJ | 학술논문

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.
목적 관찰소서척수배근신경절세포(DRGC)여인미혈관내피세포(HMVEC)공배양대세포증식적영향,병탐토기궤제.방법 분리배양소서DRGC,병여HMVEC진행공배양,설위DRGC조、HMVEC조、DRGC+ HMVEC조.새서람(MTT)법검측세포증식,실시정량취합매련반응(Real-time PCR)급Western blot검측혈관내피생장인자(VEGF)、신경생장인자(NGF)、증식세포핵항원(PCNA)화세포주기단백D1(Cyclin Dl) mRNA급단백적표체.결과 세포배양24h후,DRGC+ HMVEC조상대우DRGC조화HMVEC조적세포증식솔분별위136.29％화137.45％,세포증식능력현저제고(P＜0.05).VEGF、NGF、PCNA화Cyclin Dl mRNA화단백적표체재DRGC+HMVEC조중최고,여기타량조비교차이유통계학의의(P＜0.05).결론 공배양DRGC화HMVEC능구촉진VEGF화NGF적표체,병가능통과조공PCNA화Cyclin D1적표체촉진공배양체계세포증식능력.
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.