摘要
目的 观察吡咯喹啉醌（PQQ）对许旺细胞（Sc）的增殖作用,并探讨其对Sc c-fos、c-jun、CREB及PCNA表达的影响.方法 体外原代培养、纯化及鉴定Sc;行无血清培养细胞周期同步化,应用不同浓度PQQ（0、1、10、100、1 000、10 000 nmol/L）作用sc 72 h;流式细胞仪检测细胞周期比例;RT-PCR检测c-fos、c-jun、CREB的mRNA含量;Western blot技术检测PCNA蛋白表达.结果 PQQ处理组表现为G0/G1期细胞比例减少,S期和G2/M期细胞所占比例增加;100 nmol/L PQQ可使c-fos、c-jun、CREB mRNA含量分别增加0.33、0.42和0.52倍（P＜0.05）;PQQ浓度为1 000 nmol/L时,上述因子mRNA含量与对照组比较差异无统计学意义（P＞0.05）;PQQ浓度为10 000 nmol/L时,上述因子mRNA的含量均降低（P＜0.05）;PQQ浓度在1～100 nmol/L时,PCNA蛋白表达上调,且当PQQ浓度为100 nmol/L时PCNA蛋白上调效果最为明显,与对照组比较增加了1.17倍（P＜0.05）;当PQQ浓度为1000 nmol/L时PCNA的表达与对照组比较差异无统计学意义（P＞0.05）;当PQQ浓度为10 000 nmol/L时PCNA表达降低（P＜0.05）.结论 10～100 nmol/L PQQ可促进Sc增殖,且c-fos、c-jun、CREB、PCNA在PQQ促Sc增殖过程中表达上调.
Objective To investigate the effects of pyrroloquinoline quinine （ PQQ ） on proliferation and expression of c-fos, c-jun, CREB and PCNA in cultured Schwann cells. Methods Schwann cells were cultured and purified in vitro. The purity of Schwann cells was identified by immunofluorescence of S-100. After synchronization of cell cycle by serum-free medium, different concentration of PQQ（0,1,10,100,1 000,10 000 nmol/L ） were added into culture medium for 72 h. Flow cytometry was used to determine cell cycle. The content of c-fos, c-jun, and CREB mRNA were detected by RT-PCR, and the expression of PCNA protein was detected by Western blot. Results After PQQ treatment, the percentage of cells in G0/G1 phase decreased and the percentage of cells in S and G2/M phase increased. After treated by PQQ at concentration of 1-10 000nmol/L, content of c-fos,c-jun, CREB mRNA was increased by 0. 33 , 0. 42 and 0. 52 fold （P〈0. 05 ） . However, at concentration of 1 000 nmo1/L, there was no difference in mRNAs content when compare to control（P 〉0. 05）. And it showed a decline at concentration of 10 000 nmol/L（P〈0. 05）. PCNA protein expression was up-regulated at PQQ concentration of 1-100 nmol/L. At 100 nmol/L, the expression increased by 1.17 fold （P〈 0. 05 ） ; However, at 1 000 nmol/L, there was no difference in PCNA expression when compared to control. And 10 000 nmol/L of PQQ inhibited the expression of PCNA （P〈0. 05）. Conclusions When treated with PQQ at concentration of 10-100 nmol/L, the proliferation of Schwann cells increased and the expression of c-fos,c-jun,CREB and PCNA was up-regulated.
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2011年第4期298-303,共6页
Chinese Journal of Plastic Surgery
基金
国家自然科学基金资助项目（30600627,30570496）
武汉市青年科技晨光计划资助项目（200750731256）
关键词
吡咯喹啉醌
许旺细胞
细胞增殖
Pyrroloquinoline quinone
Schwann cell
Cell proliferation
