摘要
目的:观察运动性免疫失衡对调控骨髓早期B细胞分化发育的特异性转录因子的影响。方法:SPF级雄性SD大鼠128只,随机分为绝对安静组(Absolute control,简称AC组),运动前安静组(Control,简称A组)、运动后即刻组(Immediate简称J组)、运动后3h组(3Hours,简称3H组),进行6周递增负荷跑台运动,分别在第0、2、4、6周末用实时荧光定量PCR、Western Blot或脱钙股骨免疫组化检测其骨髓中转录因子E2A、EBF、PAX5的蛋白及基因表达。结果:6周递增负荷运动中,在第0周,各组间E2A、EBF、PAX5蛋白表达均无显著性差异(P>0.05),而J组的EBF基因表达已呈显著性下降(P<0.05);第2周,与A组相比,J组E2A、EBF蛋白表达均显著性下降(P<0.05),而PAX5蛋白仅3H组呈显著性上升(P<0.05),并且3H组仅EBF基因表达具显著性低下(P<0.05);第4周,J组EBF蛋白有非常显著性下降(P<0.01),3H组有显著性上升(P<0.05),EBF、PAX5基因表达呈显著性下降(P<0.05);第6周,PAX5蛋白在3H组有显著性上升(P<0.05)。EBF,PAX5与Pro B、Pre B数量百分比之间呈较高正性相关系数(EBF蛋白表达与Pro B、Pre B的相关性分别为0.82、0.89;PAX5蛋白表达与Pro B、Pre B的相关性分别为0.81、0.87;EBFmR-NA与Pro B、Pre B的相关性分别为0.89、0.92)。结论与建议:1)6周递增负荷运动过程中,E2A蛋白受运动的影响,波动幅度较小,EBF、PAX5则波动幅度较大。EBF、PAX5与骨髓pro B、pre B细胞数的相关度再次提示转录因子EBF、PAX5对发育中pro B、pre B细胞数量起主导调控作用。2)E2A、PAX5的蛋白与基因表达不同步,提示基因表达对蛋白质合成只是具有指导性合成的作用,还与合成酶的活性、合成底物的充足与否等多种因素有关,此外,还可能涉及基因转录后调控机制,有待进一步研究证实。3)EBFmRNA在WK0呈现波动,提示EBF基因的转录对运动刺激更为敏感,并发现EBF比上级调控因子E2A和下游调控因子PAX5对B细胞发育的主导调控作用更加关键。
Objective：The purpose of this study was to explore the effects of exercise- induced immune imbalance on the transcription factor E2A,EBF,PAX5 of early precursor B lymphocytes development in rat bone marrow.Method：128 Sprague–Dawley rats were randomly assigned to absolute control （AC）,control （A）,immediateness （J） and 3 hours （3H） after exercises,respectively.E2A,EBF,PAX5 protein and gene expression were measured by Real-time fluorescent quantitation PCR,Western Blot or decalcified femur immunohistochemisty at the end of 0 week,2nd,4th and 6th week.Result：WK 0：EBFmRNA at WK 0 J had shown a significant decrease （P〈0.05） compared with A group.WK 2：E2A,EBF protein expression at WK 2 I was significantly decreased （P〈0.05）,and PAX5 only at WK 2 3H was significantly increased （P〈0.05） compared with A group.WK 4：EBF protein expression at WK 4 J showed a very significant decrease （P〈0.01）,EBF mRNA,PAX5 mRNA was significantly decreased （P〈0.05） compared with A group.WK 6：PAX5 protein expression at WK 6 3H was significantly increased （P〈0.05）.EBF,PAX5 and Pro B,Pre B correlation coefficient （EBF ＆ Pro B,r=0.82;EBF ＆ Pre B,r=0.89;PAX5 ＆ Pro B,r=0.81;PAX5 ＆ Pre B,r=0.87;EBFmRNA ＆ Pro B,r=0.89;EBFmRNA ＆ Pre B,r=0.92）.Conclusion：1,EBF,PAX5 were more sensitive to exercise load than E2A.EBF,PAX5 protein and pro B,pre B cell mass in bone marrow to establish the correlation coefficient,once again prompted the transcription factor EBF,PAX5 were the prominent role of the dominant control of the development of pro B,pre B cells;2,EBFmRNA in WK0 showed a significant decrease.It suggests that EBF was even more crucial to the regulation of B cell development than E2A,PAX5.
出处
《体育科学》
CSSCI
北大核心
2012年第11期76-82,共7页
China Sport Science
基金
广东省自然科学基金资助项目(915106310100059)
山东体育学院重点课题(111101)
关键词
运动性免疫失衡
B淋巴细胞
发育
骨髓
转录因子
鼠
动物实验
exercise-induced immune imbalance
B lymphocytes
development
bone marrow
transcription factor；rat；animal experimen
