Shh和Fgf8介导的视黄酸对22q11.2微缺失综合征心肌细胞TBX1表达的影响  

Influences of Retioic Acid Induced by Shh and Fgf8 on TBX1 Expression in 22q11.2 Microdeletion Syndrome

作　　者：刘苗[1] 吴小艳[1] 徐佳伟[1] 金润铭[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院儿科,武汉430022

出　　处：《实用儿科临床杂志》2008年第20期1569-1572,共4页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金项目资助(30400484)

摘　　要：目的探讨Shh和Fgf8介导的视黄酸(RA)对22q11.2微缺失综合征中心肌细胞TBX1基因的调节机制。方法出生1～2 d SD乳鼠26只,分离培养乳鼠心肌细胞,48 h后用流式分选方法分选纯化心肌细胞,纯化的心肌细胞继续培养48 h。分为空白对照组;阴性对照组;处理1组:1×10-7mol/L RA;处理2组:3×10-7mol/L RA;处理3组:5×10-7mol/L RA。采用RT-PCR和Western blot方法分别检测乳鼠心肌细胞Shh、Fgf8 mRNA和蛋白相对量表达的变化。结果Shh和Fgf8在乳鼠心肌细胞有少量的表达,给予RA刺激后,与空白对照组比较,3×10-7mol/L RA使细胞内Shh mRNA和蛋白水平分别增加1.51倍和1.10倍(t=11.328,15.598 Pa<0.05),5×10-7mol/LRA使细胞内Shh mRNA和蛋白水平分别增加2.21倍和2.38倍(t=13.761,18.982Pa<0.05);3×10-7mol/L RA使细胞内Fgf8 mRNA和蛋白水平分别增加2.50倍和0.80倍(t=14.368,24.226 Pa<0.05),而5×10-7mol/LRA使细胞内Fgf8 mRNA和蛋白水平分别增加3.48倍和2.04倍(t=23.926,26.364 Pa<0.05)。结论RA可以上调乳鼠心肌细胞Shh和Fgf8的表达,且呈剂量依赖效应。Shh和Fgf8参与了RA对心肌细胞TBX1的调节过程。Objective To explore the regulatory mechanism of retioic acid induced by Shh and Fgf8 on the TBX1 expression in myocardial cells in the chromosome 22q11.2 microdelction syndrome. Methods Twenty - six Sprague - Dawley neonatal rats horned 1 - 2 days were obtained,and ventricular cardiomyocytes were isolated from neonatal rats and cultured for 48 h, then sorted the cardiomyocytes by means of fluorescence- activated cell corting（FACS）. The sorted cells were continuously cultured for 48 h and divided into 5 groups：control group; negative control group;RA stimulation 1 group：1 ×10 ^-7 mol/L RA;RA stimulation 2 group：3 × 10^-7 mol/L RA;RA stimulation 3 group：5 × 10^- 7 mol/L RA. The expressions of Shh and Fgf8 in the levels of mRNA and protein in myocardial cells of neonatal rat were measured using reverse transcription polymerase chain reaction and Western blot technique. Results There was basal expressions of Shh and Fgf8 in control group. After RA treatment, compared with control group, the expressions of Shh mRNA and protein in myocardial cells of neonatal rat were up - regulated by 1.51 -fold and 1.10- fold（t =11.328,15.598 Pa 〈0. 05,respectively） under 3 × 10^-7 mol/L RA,and 2.21 -fold and 2.38 - fold（t = 13. 761,18. 982 Pa〈 0.05） respectively under 5 × 10 ^-7 mol/L RA. Meanwhile, compared with control group, the expressions of Fgf8 in the level of mRNA and protein in myocardial cells of neonatal rat were up - regulated by 2.50 - fold and 0.8 - fold（ t = 14. 368, 24.226 Pa 〈0.05） respectively simulated by 3 ×10^-7 mol/L RA,and 3.48 -fold and 2.04 -fold（t =23.926,26.364 Pa 〈0.05） respectively simulated by 5 × 10 ^-7 mol/L RA. Conclusions Retioic acid can up - regulate the expressions of Shh and Fgf8 in myocardial cells of neonatal rat,and that is dose- dependent. Shh and Fgf8 are involved in the regulation process of retioic acid on the TBX1 expression.

关 键 词：视黄酸 TBX1基因 Shh基因 Fgf8基因 

分 类 号：R725.9[医药卫生—儿科]