CAJ | 학술논문

目的：检测维甲酸（Retinoic acid，RA）诱导小鼠胚胎腭裂模型中胎鼠舌体发育过程中肌相关 microRNAs（MyomiRs）、成肌调节因子（Myogenic regulatory factors，MRFs）以及 Pax 基因的表达变化，探究 MyomiRs 在舌肌分化过程中的调控作用，推测RA 致胎鼠腭裂伴发舌异常的可能机制。方法：建立 RA 诱导小鼠胚胎腭裂模型，分别在 E13．5、E14．5、E15．5收集胎鼠舌体组织，用 SYBR GreenⅠ实时定量 PCR 检测舌体中 MRFs 和 Pax 基因的表达；用 TaqMan 探针实时定量 PCR 检测舌体中 MyomiRs 的表达。结果：胎鼠舌体发育过程中，正常组 miR-1和 miR-206相对表达量均持续上升，RA 诱导组二者变化趋势与正常组相似，但相对表达量均低于正常组，miR-1的结果在 E14．5和 E15．5具有统计学意义（P ＜0．01），miR-206的结果在 E13．5具有统计学意义（P ＜0．05）。正常组和 RA 诱导组胎鼠舌体中 MyoD 和 Myf5相对表达量都在 E14．5达到峰值，随后下降。RA 诱导组 MyoD 的表达在 E14．5显著低于正常组（P ＜0．05），在 E15．5显著高于正常组（P ＜0．01）；RA 诱导组 Myf5的表达在 E15．5显著低于正常组（P ＜0．05）。正常组和 RA 诱导组胎鼠舌体中 Pax3表达均在 E14．5达到峰值，Pax7表达均在 E15．5达到峰值。RA 诱导组 Pax3的表达在 E14．5显著高于正常组（P ＜0．05）；Pax7的表达则在 E13．5显著高于正常组（P ＜0．01）。结论：在舌肌分化过程以及RA 诱导腭裂胎鼠的舌发育异常中，miR-1／miR-206与 Pax3／Pax7及 Myf5／MyoD 的表达趋势具有相关性。RA 可能通过下调 miR-1／miR-206而靶向上调 Pax3／Pax7，进而下调 MyoD ／Myf5表达，从而抑制舌肌分化，导致舌肌发育异常。
목적：검측유갑산（Retinoic acid，RA）유도소서배태악렬모형중태서설체발육과정중기상관 microRNAs（MyomiRs）、성기조절인자（Myogenic regulatory factors，MRFs）이급 Pax 기인적표체변화，탐구 MyomiRs 재설기분화과정중적조공작용，추측RA 치태서악렬반발설이상적가능궤제。방법：건립 RA 유도소서배태악렬모형，분별재 E13．5、E14．5、E15．5수집태서설체조직，용 SYBR GreenⅠ실시정량 PCR 검측설체중 MRFs 화 Pax 기인적표체；용 TaqMan 탐침실시정량 PCR 검측설체중 MyomiRs 적표체。결과：태서설체발육과정중，정상조 miR-1화 miR-206상대표체량균지속상승，RA 유도조이자변화추세여정상조상사，단상대표체량균저우정상조，miR-1적결과재 E14．5화 E15．5구유통계학의의（P ＜0．01），miR-206적결과재 E13．5구유통계학의의（P ＜0．05）。정상조화 RA 유도조태서설체중 MyoD 화 Myf5상대표체량도재 E14．5체도봉치，수후하강。RA 유도조 MyoD 적표체재 E14．5현저저우정상조（P ＜0．05），재 E15．5현저고우정상조（P ＜0．01）；RA 유도조 Myf5적표체재 E15．5현저저우정상조（P ＜0．05）。정상조화 RA 유도조태서설체중 Pax3표체균재 E14．5체도봉치，Pax7표체균재 E15．5체도봉치。RA 유도조 Pax3적표체재 E14．5현저고우정상조（P ＜0．05）；Pax7적표체칙재 E13．5현저고우정상조（P ＜0．01）。결론：재설기분화과정이급RA 유도악렬태서적설발육이상중，miR-1／miR-206여 Pax3／Pax7급 Myf5／MyoD 적표체추세구유상관성。RA 가능통과하조 miR-1／miR-206이파향상조 Pax3／Pax7，진이하조 MyoD ／Myf5표체，종이억제설기분화，도치설기발육이상。
Objective:To research the mechanisms of the muscle-specific microRNAs in the tongue′s differentiation of the retinoic acid induced tongue dysplasia by measuring the expression of MyomiRs,MRFs and Pax3 /Pax7.Methods:A retinoic acid induced tongue dysplasia mouse model was established.The tongues were collected at E13.5,E14.5 and E15.5.The expression levels of MyomiRs, MRFs and Pax3 /Pax7 were detected by real-time quantitative PCR.Results:During the tongue development,the expression of miR-1 and miR-206 kept on rising both in the tongues of RA-treated mice and normal ones.While the expression of miR-1 and miR-206 in the RA-treated mice′s tongues was less than in the normal ones.The results of miR-1 at E14.5 and E15.5 had statistical significance (P <0.01) and the results of miR-206 at E13.5 had statistical significance (P <0.05).The expression of MyoD and Myf5 in the normal and RA-treated mice′s tongues reached the peak at E14.5 then decreased.At E13.5 and E14.5,the expression of MyoD in the RA-treated mice′s tongues was less than the controls.But at E15.5,the expression of MyoD in the RA-treated mice′s tongues was higher (P <0.01).And at E14.5 and E15.5,the expression of Myf5 in the RA-treated mice′s tongues was lessthan the normal subjects.During the tongue myo-genesis of normal and RA-treated mice the expression of Pax3 in the tongue reached the peak at E14.5,and the expression of Pax7 in both of them reached the peak at E15.5.Compared to the normal mice,the expression of Pax3 was higher at E14.5 (P <0.05),and the ex-pression of Pax7 was higher at E13.5 (P <0.01)in the RA-treated mice′s tongues.Conclusions:During the tongue myogenesis,miR1 /miR-206,Pax3 /Pax7 and Myf5 /MyoD were associated.In the retinoic acid induced tongue dysplasia,the correlation still existed.RA may down-regulate miR-1 /miR-206 which targeted on Pax3 /Pax7,following down-regulate the expression of MyoD /Myf5 to inhibit the myogen-esis in tongues of RA-treated mice,resulted in tongue dysplasia.