先天性肥厚性幽门狭窄与神经生长因子及其受体表达的相关性研究

Correlation analysis between congenital hypertrophic pyloric stenosis and the nerve growth factor and its receptor expressions

目的研究先天性肥厚性幽门狭窄(CHPS)与神经生长因子(NGF)及其受体(TrkA与p75NTR)表达的相关性。方法用N-硝基-左旋精氨酸甲酯(L-NAME)构建CHPS大鼠模型。对照组予以等量0.9%Na Cl,尾静脉注射。实验组予以20 mg·kg^(-1)·d^(-1)L-NAME,尾静脉注射。2组均从妊娠第2天开始给药至第21天。用HE染色后,在显微镜下测量幽门肌层厚度;用定量实时荧光聚合酶链反应检测幽门组织中的NGF及其受体TrkA与p75NTR的mRNA表达水平;用Western Blot法检测幽门组织中的NGF及其受体TrkA与p75NTR的蛋白表达水平。结果实验组和对照组的幽门肌层厚度分别为(0.51±0.11)和(0.29±0.09)mm,NGF mRNA分别为(2.51±0.39)和(1.01±0.02),TrkA mRNA分别为(0.48±0.22)和(1.02±0.01),p75NTR mRNA分别为(0.52±0.25)和(1.01±0.01),NGF蛋白分别为(0.43±0.05)和(0.23±0.04),TrkA蛋白分别为(0.21±0.14)和(0.53±0.16),p75NTR蛋白分别为(0.22±0.19)和(0.71±0.11),差异均有统计学意义(均P<0.05)。结论幽门组织中NGF、TrkA与p75NTR表达的变化可能与CHPS发病存在相关性。

Objective To investigate the correlations between congenital hypertrophic pyloric stenosis（ CHPS） and nerve growth factor（ NGF）and its receptor（ TrkA and p75 NTR） expressions. Methods CHPS model was established by administering N-nitro-L-arginine methylester（ L-NAME）. Control group was treated with equal 0. 9% Na Cl,tail vein injection. Test group was given L-NAME 20 mg ·kg^（-1）·d^（-1）,tail vein injection. Two groups were administered from day 2 to day 21. After HE staining,pyloric muscle thickness was measured with microscope; the expression levels of NGF,TrkA and p75 NTR mRNA were measured with quantitative reverse transcription and polymerase Chain reaction; the expression levels of NGF,TrkA and p75 NTR protein were measured with Western Blotting. Results The main indexes of test and control groups were compared： pyloric muscle thickness were（ 0. 51 ± 0. 11） and（ 0. 29 ± 0. 09） mm, NGF mRNA were（ 2. 51 ± 0. 39） and（ 1. 01 ± 0. 02）,TrkA mRNA were（ 0. 48 ± 0. 22） and（ 1. 02 ± 0. 01）,p75 NTR mRNA were（ 0. 52 ± 0. 25） and（ 1. 01 ± 0. 01）,NGF protein were（ 0. 43 ± 0. 05） and（ 0. 23 ± 0. 04）,TrkA protein were（ 0. 21 ± 0. 14） and（ 0. 53 ± 0. 16）,p75 NTR protien were（ 0. 22 ± 0. 19） and（ 0. 71 ± 0. 11）,the differences were statistically significant（ all P〈0. 05）. Conclusion The changes of NGF,TrkA and p75 NTR expression were likely associated with CHPS pathology.