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中华胸部外科电子杂志 ›› 2017, Vol. 04 ›› Issue (01) : 1 -6. doi: 10.3877/cma.j.issn.2095-8773.2017.01.01

所属专题: 文献

论著

内皮素受体在人食管下括约肌上的分布及其作用
张珂1, 刘俊峰2,(), 何士骄3, 樊崇利4, 吴艳刚4, 包永松4, 宋晓辉4   
  1. 1. 071000 保定,河北大学附属医院胸外科
    2. 050019 石家庄,河北医科大学第四医院胸外科
    3. 071100 保定市清苑区人民医院肿瘤科
    4. 071100 保定市清苑区人民医院骨科
  • 收稿日期:2016-11-05 出版日期:2017-02-28
  • 通信作者: 刘俊峰
  • 基金资助:
    国家自然科学基金(30371413)

Distribution and role of endothelin receptors in human lower esophageal sphincter

Ke Zhang1, Junfeng Liu2,(), Shijiao He3, Chongli Fan4, Yangang Wu4, Yongsong Bao4, Xiaohui Song4   

  1. 1. Department of Thoracic Surgery, Affiliated Hospital of Hebei University, Baoding 071000, China
    2. Department of Thoracic Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050019, China
    3. Department of Oncology, The People’s Hospital of Qingyuan District of Baoding City, Baoding 071100, China
    4. Department of Orthopaedics, The People’s Hospital of Qingyuan District of Baoding City, Baoding 071100, China
  • Received:2016-11-05 Published:2017-02-28
  • Corresponding author: Junfeng Liu
  • About author:
    Corresponding author: Liu Junfeng, Email:
引用本文:

张珂, 刘俊峰, 何士骄, 樊崇利, 吴艳刚, 包永松, 宋晓辉. 内皮素受体在人食管下括约肌上的分布及其作用[J]. 中华胸部外科电子杂志, 2017, 04(01): 1-6.

Ke Zhang, Junfeng Liu, Shijiao He, Chongli Fan, Yangang Wu, Yongsong Bao, Xiaohui Song. Distribution and role of endothelin receptors in human lower esophageal sphincter[J]. Chinese Journal of Thoracic Surgery(Electronic Edition), 2017, 04(01): 1-6.

目的

通过检测各内皮素受体(ETR)亚型在人食管下括约肌(LES)上,特别是套索纤维和钩状纤维上的分布规律,探讨内皮素对LES收缩和舒张功能的作用。

方法

在罹患食管中上段癌患者的术后标本上,取LES的套索纤维和钩状纤维,并分别以食管下段和胃底部的环形肌肉作为对照。采用蛋白印迹法(Western Blot)、RT-PCR和半定量PCR(semi-qPCR)分析ETR在人LES上的分布规律。

结果

ETR的A亚型(ETA)和B亚型(ETB)被确认都存在于LES上。ETR两个受体亚型的蛋白质和mRNA表达水平比较,差异均有统计学意义(F=533.841,P<0.01;F=72.291,P<0.01)。ETA显著高于ETB表达量(约为5∶1),差异有统计学意义;而同一受体亚型在钩状纤维(C)、套索纤维(S)、食管下端环形肌(E)、胃底环形肌(G)4个位点上的表达量比较差异无统计学意义(F=0.398,P=0.5329)。

结论

ET受体,特别是ETA受体亚型在人LES的舒张、收缩功能调节中发挥着重要作用,是食管运动障碍性疾病治疗的靶向受体。

Objective

To detect the distribution of endothelin receptors(ETR) in the human lower esophageal sphincter (LES), especially on the clasp and sling smooth muscle fibers of the LES, and investigate the effects of endothelins on the contraction and diastole of LES.

Methods

The sling and clasp tissues from LES were obtained from esophageal carcinoma patients, and the circular muscle tissues from the esophagus and gastric fundus were served as controls. Western Blot, RT-PCR and semi-qPCR were employed to determine the distribution of ETR in human LES.

Results

Endothelin receptor type A (ETA) and endothelin receptor type B (ETB) were found in human LES. There were significant differences in the expression of protein and mRNA between two isoforms of ETR (F=533.841, P<0.01; F=72.291, P<0.01). The expression level of ETA was significantly higher than that of ETB(about 5: 1). For distribution of each isoform, there was no significant difference in the expression level among four sites of clasp fibers(C), sling fibers(S), lower esophageal sphincter(E) and gastric fundus circular muscle(G)(F=0.398, P=0.5329).

Conclusions

ETRs contribute to the adjustment of contraction and diastole of human LES, especially the ETA. ETRs are the potent therapeutic targets for esophageal mobility diseases.

图1 食管胃结合部标本沿胃大弯切开后固定于蜡盘上,锐性分离掉掉黏膜和黏膜下组织
表1 RT-PCR引物
表2 实时定量PCR引物
图2 同一肌条上,ETA和ETB的表达量有显著差异,且4个检测位点情况类似。N:空白对照;β-actin:内参
图3 检测的4个肌条上,同一受体的表达量无显著差异。C:钩状纤维(clasp);S:套索纤维(sling) ;E:食管下端环形肌(esophagus);G:胃底环形肌(gastric fundus)
图4 qPCR各扩增曲线分离
图5 qPCR检测显示,同一肌条上2个受体亚型表达量有显著差异(F=72.291,P=0.000);同一受体亚型在检测的各位点间无统计学差异(F=0.398,P=0.533)。C:钩状纤维(clasp);S:套索纤维(sling) ;E:食管下端环形肌(esophagus);G:胃底环形肌(gastric fundus)
图6 Western Blot 检测结果。在同一肌条上,两种亚型受体表达量有显著差异(F=533.841,P<0.01);同一受体亚型在4种不同肌条上的表达量无统计学差异(F= 0.398,P=0.755)。C:钩状纤维(clasp);S:套索纤维(sling) ;E:食管下端环形肌(esophagus);G:胃底环形肌(Gastric fundus)
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