维A酸通过上调脑源性神经营养因子促进小鼠胚胎干细胞向神经细胞分化的研究  

Retinoic acid promotes differentiation of mouse embryonic stem cells into neural cells by up-regulating brain derived neurotrophic factor

作　　者：宋祖荣[1] 陈涛 SONG Zu-rong;CHEN Tao(College of Pharmacy,Anhui Xinhua University,Hefei 230088,Anhui Province,China)

机构地区：[1]安徽新华学院药学院,安徽合肥230088

出　　处：《中国临床药理学杂志》2022年第6期491-494,503,共5页The Chinese Journal of Clinical Pharmacology

基　　金：安徽省教育厅自然研究重点课题基金资助项目(KJ2018A0583);安徽新华学院校级自然研究重点课题基金资助项目(2017zr010);国家级创新创业训练计划创新训练基金资助项目(201712216034)。

摘　　要：目的 探讨维A酸对小鼠胚胎干细胞发生神经分化的影响及机制。方法 用1μmol·L^(-1)维A酸对小鼠胚胎干细胞E14Tg2A.4处理0和5 d,分别标记为对照组和实验组。用脂质体法将si-NC、si-BDNF、pcDNA、pcDNA-BDNF、si-BDNF+NS(转染si-BDNF并用0.9%NaCl处理5 d)、si-BDNF+RA(转染si-BDNF并用1μmol·L^(-1)维A酸处理5 d)转染至E14Tg2A.4细胞,转染成功后分别标记为si-NC组、si-BDNF组、pcDNA组、pcDNA-BDNF组、si-BDNF+NS组、si-BDNF+RA组。用实时荧光定量聚合酶链反应法检测sox2、nestin和tuj1 mRNA的表达水平,用Western blot法检测脑源性神经营养因子(BDNF)蛋白的表达水平。结果 对照组、实验组、si-NC组、si-BDNF组、pcDNA组、pcDNA-BDNF组、si-BDNF+NS组和si-BDNF+RA组的sox2 mRNA相对表达量分别为1.00±0.06,0.16±0.01,1.00±0.08,0.32±0.03,1.00±0.07,0.32±0.03,1.01±0.07和0.72±0.06,nestin mRNA相对表达量分别为1.00±0.08,1.94±0.14,1.01±0.06,1.98±0.16,1.01±0.06,1.98±0.16,1.02±0.08和1.32±0.09,tuj1 mRNA相对表达量分别为1.01±0.08,2.64±0.23,1.01±0.09,1.98±0.14,0.99±0.09,1.98±0.14,1.02±0.07和1.61±0.12,BDNF蛋白相对表达量分别为1.00±0.09,3.16±0.28,1.00±0.07,0.35±0.03,1.01±0.06,3.86±0.34,1.02±0.09和1.81±0.13。实验组的上述指标与对照组比较,si-BDNF组的上述指标与si-NC组比较,pcDNA-BDNF组的上述指标与pcDNA组比较,si-BDNF+RA组的上述指标与si-BDNF+NS组比较,差异均有统计学意义(均P<0.05)。结论 维A酸可通过上调BDNF表达,促进小鼠胚胎干细胞发生神经分化。Objective To investigate the effect and mechanism of retinoic acid on neural differentiation of mouse embryonic stem cells.Methods Mouse embryonic stem cells E14Tg2A.4 were treated with 1μmol·L^(-1)retinoic acid for 0 and 5 days,and they were labeled as control and experimental groups.Si-NC,si-BDNF,pcDNA,pcDNA-BDNF,si-BDNF+NS (transfected with si-BDNF and treated with0.9%NaCl for 5 days) and si-BDNF+RA (transfected with si-BDNF and treated with 1μmol·L^(-1)retinoic acid for 5 days) were transfected into E14Tg2A.4 cells by lipofectamine.After successful transfection,they were labeled as si-NC,si-BDNF,pcDNA,pcDNA-BDNF,si-BDNF+NS and si-BDNF+RA groups.Quantitative real-time transcription-polymerase chain reaction was used to detect the expression levels of sox2,nestin and tuj1 mRNA,and the expression level of BDNF protein in cells was determined by Western blot.Results The relative expression levels of sox2 mRNA in the control,experimental,si-NC,si-BDNF,pcDNA,pcDNA-BDNF,si-BDNF+NS and siBDNF+RA groups were 1.00±0.06,0.16±0.01,1.00±0.08,0.32±0.03,1.00±0.07,0.32±0.03,1.01±0.07 and 0.72±0.06,the relative expression levels of nestin mRNA were 1.00±0.08,1.94±0.14,1.01±0.06,1.98±0.16,1.01±0.06,1.98±0.16,1.02±0.08 and 1.32±0.09,the relative expression levels of tuj1 mRNA were 1.01±0.08,2.64±0.23,1.01±0.09,1.98±0.14,0.99±0.09,1.98±0.14,1.02±0.07and 1.61±0.12,the relative expression levels of BDNF protein were 1.00±0.09,3.16±0.28,1.00±0.07,0.35±0.03,1.01±0.06,3.86±0.34,1.02±0.09 and 1.81±0.13.The differences were statistically significant between experimental group and control group,between si-BDNF group and si-NC group,between pcDNA-BDNF group and pcDNA group,between si-BDNF+RA group and si-BDNF+NS group (all P<0.05).ConclusionRetinoic acid can promote the neural differentiation of mouse embryonic stem cells by up-regulating the expression of BDNF.

关 键 词：维A酸 脑源性神经营养因子 小鼠胚胎干细胞 神经分化 

分 类 号：R97[医药卫生—药品]