外源性神经生长因子对雪旺细胞中NF-κB mRNA表达的影响

Influence of exogenous nerve growth factor on expression of NF-κB mRNA of Schwann cells

目的：观察外源性神经生长因子（NGF）对雪旺细胞（SCs）中核转录因子κB（NF-κB）mRNA表达的影响。方法体外培养SCs，用S-100免疫组化染色鉴定细胞纯度。取第4代优质细胞，随机分为实验组（ NGF）、阻断组（NGF＋anti-NGFR）、对照组（PBS），提取各组总mRNA，分别用紫外分光光度计、1．5％琼脂糖凝胶电泳观察结果，通过实时荧光定量PCR检测NF-κB mRNA的表达。结果体外培养的SCs与倒置显微镜下观察到的活体细胞形态一致，S-100免疫组化染色显示，SCs纯度达到91％。紫外分光光度计观察A260/A280为1．8～2．2，1．5％琼脂糖凝胶电泳可见28、18 s两条清晰条带，证实提取的mRNA质量良好。实时荧光定量PCR检测发现，与对照组、阻断组相比，实验组NF-κB mRNA的表达显著增高（P均＜0．01）。结论外源性NGF可以上调SCs中NF-κB mRNA的表达。

Objective To observe the influence of exogenous nerve growth factor （ NGF） on the expression of nuclear transcription factor-κB （NF-κB） mRNA of Schwann cells （SCs）.Methods Schwann cells were isolated and cultured in vitro.The positive cells were counted by using S-100 immunohistochemical staining .The fourth generation of high-quality cells were randomly divided into the experimental group （NGF）, blocking group （NGF＋anti-NGFR） and control group （PBS）.The total mRNA in each group were extracted and observed by ultraviolet spectrophotometer and 1.5%agarose gel electrophoresis , respectively , and the expression of NF-κB mRNA were detected by real-time fluorescence quantitative PCR.Results The living cell morphologies of SCs cultured in vitro were consistent with those observed under inverted microscope.S-100 immunohistochemical staining showed that the purity of SCs was 91%.Ultraviolet spectrophotometer observed that A260/A280 was 1.8-2.2, 1.5% agarose gel electrophoresis showed that there were 28 and 18 s clear bands, which confirmed the quality of extracted mRNA was good .Real time fluorescence quantitative PCR in detection of expres-sion of NF-κB mRNA showed that when compared with the control group and blocking group , the expression of NF-κB mR-NA in the experimental group was significantly increased （all P〈0.01）.Conclusions Exogenous NGF can up-regulate the expression of NF-κB mRNA.