PGP9.5、SYn和α-actin在先天性肛门直肠畸形动物模型盆底肌的表达及意义
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摘要
目的检测PGP9.5、SYn和α-actin在胎鼠正常盆底肌和肛门直肠畸形(ARMs)中的蛋白表达,探讨其表达意义。方法通过全反式维甲酸(ATRA)诱导大鼠胚胎产生ARMs动物模型,应用免疫组化方法检测正常与ARMs胎鼠直肠末端组织PGP9.5、SYn、α-actin在E16、18、20胎鼠盆底肌发育过程中的表达。结果正常组共产胎鼠91只,未见畸形、死胎、吸收胎;ATRA诱导大鼠共产胎鼠126只,除无肛畸形外,短尾、无尾畸形58例,存在显性脊柱裂、脑膜膨出等神经发育缺陷有29例,ARMs发生率为74.60%。E16、18、20胎鼠盆底肌:正常组PGP9.5、SYn和α-actin呈阳性表达;ARMs组PGP9.5、SYn和α-actin呈弱阳性表达,分布稀疏。PGP9.5、SYn强阳性表达率除E16 2组间差异无统计学意义外(P﹥0.05),E18、20差异有统计学意义(P<0.05)。α-actin在E16、18、20两种组织间强阳性表达率,差异有统计学意义(P<0.05)。结论 PGP9.5、SYn和α-actin分布异常是ARMs动物模型直肠末端壁的重要病理改变之一。
Objective To explore the PGP9. 5,SYn and α- actin protein expression in the pelvic floor muscles and ARMs in normal fetuses,and its meaning of expression. Methods ARMs animal models were produced by all trans retinoic acid( ATRA)- induced rat embryos,using immunohistochemical methods to detect PGP9. 5,SYn,α- actin expression in fetal pelvic floor muscle of normal and anorectal malformation fetal embryos in E16,18,20. Results Normal group produced a total of 91 fetuses,no malformation,stillbirth,fetal absorption. ATRA induced rats produced a total of 126 fetuses,except for anorectal malformation,and the number of short-tailed and no tail deformity was 58. 29 cases got dominant spinal bifida or neurodevelopmental defects meninges. The incidence of ARMs was 74. 60%; Pelvic floor muscles of E16,18,20 fetal: the expression of PGP9. 5,SYn and α- actin of normal group was positive; the expression of PGP9. 5,SYn and α- actin of ARMs group were weakly positive,and distributed sparse. The strong positive expression rate of PGP9. 5,SYn between the two groups was not statistically significant for E16( P > 0. 05),and the differences of E18,20 were statistically significant( P < 0. 05). The strong positive expression rate of α- actin between the two groups of E16,18,20 was statistically significant( P < 0. 05). Conclusion The abnormal distribution of PGP9. 5,SYn and α- actin of terminal rectum wall is one of the important pathological changes in animal models of ARMs.
Objective To explore the PGP9. 5,SYn and α- actin protein expression in the pelvic floor muscles and ARMs in normal fetuses,and its meaning of expression. Methods ARMs animal models were produced by all trans retinoic acid( ATRA)- induced rat embryos,using immunohistochemical methods to detect PGP9. 5,SYn,α- actin expression in fetal pelvic floor muscle of normal and anorectal malformation fetal embryos in E16,18,20. Results Normal group produced a total of 91 fetuses,no malformation,stillbirth,fetal absorption. ATRA induced rats produced a total of 126 fetuses,except for anorectal malformation,and the number of short-tailed and no tail deformity was 58. 29 cases got dominant spinal bifida or neurodevelopmental defects meninges. The incidence of ARMs was 74. 60%; Pelvic floor muscles of E16,18,20 fetal: the expression of PGP9. 5,SYn and α- actin of normal group was positive; the expression of PGP9. 5,SYn and α- actin of ARMs group were weakly positive,and distributed sparse. The strong positive expression rate of PGP9. 5,SYn between the two groups was not statistically significant for E16( P > 0. 05),and the differences of E18,20 were statistically significant( P < 0. 05). The strong positive expression rate of α- actin between the two groups of E16,18,20 was statistically significant( P < 0. 05). Conclusion The abnormal distribution of PGP9. 5,SYn and α- actin of terminal rectum wall is one of the important pathological changes in animal models of ARMs.
引文
[1]李康.正常及先天性直肠肛门畸形大鼠盆底结构的磁共振扫描及三维结构重建[D].武汉:华中科技大学,2013.
[2]赫文波.直肠肛门畸形远端神经节细胞分布及GDNF、GFRal、RET表达情况研究[D].天津:天津医科大学,2013.
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[5]Venkatesh B,Tay BH,Elgar G,et al.Isolation characterization and evolution of nine pufferfish(fugu rubripes)actin genes[J].J Mol Biol,1996,259:655.
[6]姜霁哲.先天性肛门直肠畸形术后排便功能影响因素研究[D].银川:宁夏医科大学,2013.
[7]刘燕雄,单振潮,杜勇.全反式维甲酸诱导大鼠肠神经系统发育异常的实验研究[J].宁夏医科大学学报,2012,3(34):234-236.
[8]贾慧敏,陈青江,张涛,等.Cajal间质细胞及突触素在肛门直肠畸形动物模型末端直肠壁的表达及意义[J].中国医科大学学报,2009,1(2):36.
[9]毛羽晨,刘远梅,金祝,等.先天性肛门直肠畸形胎鼠直肠末端NGF及受体Trk A与P75的表达[J].中华小儿外科杂志,2012,33(11):854-858.
[10]Tsehematsch M,Klotz M,Probst C,et al.Synaptophysin is an autoantigen in paraneoplastic[J].Neuropathy J Neuroimmunol,2008,197(1):81-86.
[11]孔萌,刘远梅,金祝,等.嘌呤受体亚型(P2Y2)在肛门直肠畸形动物模型直肠末端的表达[J].遵义医学院学报,2014,6(37):308-311.
[12]李龙,张金哲.Pena手术及盆底肌肉折叠术在肛门闭锁术后大便失禁中的应用[J].中华小儿外科杂志,1999,20(5):278.
[13]李苗苗.盆底骨骼肌细胞受力前后的超微结构变化[D].太原:山西医科大学,2013.
[14]郭俊斌,李昭铸.大鼠无肛畸形形成的胚胎机制和α-SMA表达的研究[J].中国优生与遗传杂志,2012,12(20):32-34.
[15]张曦,岳峰,张炎,等.全反式维甲酸致先天性肛门直肠畸形胎鼠盆底肌肌肉、神经发育形态学的实验研究[J].宁夏医学杂志,2014,4(36):289-291.
[16]孔萌,刘远梅.先天性肛门直肠畸形的治疗[J].遵义医学院学报,2013,6(36):285-287.
[2]赫文波.直肠肛门畸形远端神经节细胞分布及GDNF、GFRal、RET表达情况研究[D].天津:天津医科大学,2013.
[3]Sidebotham EL,Woodward MN,Kenny SE,et al.As-sessment of protein gene product 9.5 as a marker ofneural crest derived precursor cells in the developingenteric nervous system[J].Pediatr Surg Int,2001,17:304-307.
[4]Vanden Berghe P Klingauf J.Spatial organization and dynamic properties of neurotransmitter release sites in the enteric nervous system[J].Neurosci,2007,145(1):88-99.
[5]Venkatesh B,Tay BH,Elgar G,et al.Isolation characterization and evolution of nine pufferfish(fugu rubripes)actin genes[J].J Mol Biol,1996,259:655.
[6]姜霁哲.先天性肛门直肠畸形术后排便功能影响因素研究[D].银川:宁夏医科大学,2013.
[7]刘燕雄,单振潮,杜勇.全反式维甲酸诱导大鼠肠神经系统发育异常的实验研究[J].宁夏医科大学学报,2012,3(34):234-236.
[8]贾慧敏,陈青江,张涛,等.Cajal间质细胞及突触素在肛门直肠畸形动物模型末端直肠壁的表达及意义[J].中国医科大学学报,2009,1(2):36.
[9]毛羽晨,刘远梅,金祝,等.先天性肛门直肠畸形胎鼠直肠末端NGF及受体Trk A与P75的表达[J].中华小儿外科杂志,2012,33(11):854-858.
[10]Tsehematsch M,Klotz M,Probst C,et al.Synaptophysin is an autoantigen in paraneoplastic[J].Neuropathy J Neuroimmunol,2008,197(1):81-86.
[11]孔萌,刘远梅,金祝,等.嘌呤受体亚型(P2Y2)在肛门直肠畸形动物模型直肠末端的表达[J].遵义医学院学报,2014,6(37):308-311.
[12]李龙,张金哲.Pena手术及盆底肌肉折叠术在肛门闭锁术后大便失禁中的应用[J].中华小儿外科杂志,1999,20(5):278.
[13]李苗苗.盆底骨骼肌细胞受力前后的超微结构变化[D].太原:山西医科大学,2013.
[14]郭俊斌,李昭铸.大鼠无肛畸形形成的胚胎机制和α-SMA表达的研究[J].中国优生与遗传杂志,2012,12(20):32-34.
[15]张曦,岳峰,张炎,等.全反式维甲酸致先天性肛门直肠畸形胎鼠盆底肌肌肉、神经发育形态学的实验研究[J].宁夏医学杂志,2014,4(36):289-291.
[16]孔萌,刘远梅.先天性肛门直肠畸形的治疗[J].遵义医学院学报,2013,6(36):285-287.