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目的:探讨食管下段平滑肌中是否表达Kir3及其各亚型之间表达的差异性,同时明确食管环形平滑肌(CM)与纵行平滑肌(LM)之间表达的区别。方法:根据24h食管pH值检测、食管镜检查及HE染色结果,分辨、筛选正常人食管平滑肌,按人Kir3.1～3.4序列设计并合成各自的高效引物,利用RT-PCR、蛋白质印迹法检测人食管平滑肌细胞中Kir3的表达及其亚型Kir3.1～3.4表达的差异。结果:在人食管LM、CM层平滑肌细胞中mRNA的表达分别为0.121±0.015及0.124±0.017(Kir3.2)、0.255±0.018及0.295±0.028(Kir3.3)和0.685±0.040及0.693±0.037(Kir3.4)。蛋白表达分别为0.053±0.010及0.068±0.009(Kir3.2)、0.160±0.021及0.192±0.032(Kir3.3)和0.488±0.040及0.504±0.033(Kir3.4)。Kir3.4表达最丰富,与Kir3.2与Kir3.3的表达差异有统计学意义,P<0.05;LM和CM组的表达差异无统计学意义,P>0.05。结论:人食管平滑肌细胞仅表达Kir3中的Kir3.2、Kir3.3和Kir3.4,其中Kir3.4的表达最丰富;人食管LM和CM Kir3.2、Kir3.3和Kir3.4的表达无显著差异性。
Objective: To investigate whether the expression of Kir3 and its subtypes in the lower esophageal smooth muscle express the difference between the expression of esophageal ring smooth muscle (CM) and longitudinal smooth muscle (LM). Methods: According to 24h esophageal pH value examination, esophagoscopy and HE staining results, the normal human esophageal smooth muscle was identified and screened. According to Kir3.1 ~ 3.4 sequences of human Kir3.1 ~ 3.4, efficient primers were designed and synthesized. RT-PCR and Western blotting The Expression of Kir3 in Human Esophageal Smooth Muscle Cells and the Expression of Kir3.1 ~ 3.4 in Its Subtypes. Results: The mRNA expression of human smooth muscle cells in LM and CM of human esophagus was 0.121 ± 0.015 and 0.124 ± 0.017 (Kir3.2), 0.255 ± 0.018 and 0.295 ± 0.028 (Kir3.3) and 0.685 ± 0.040 and 0.693 ± 0.037 (Kir3.4). Protein expression was 0.053 ± 0.010 and 0.068 ± 0.009 (Kir3.2), 0.160 ± 0.021 and 0.192 ± 0.032 (Kir3.3) and 0.488 ± 0.040 and 0.504 ± 0.033 (Kir3.4), respectively. The expression of Kir3.4 was the most abundant, with significant difference between Kir3.2 and Kir3.3 expression (P <0.05). There was no significant difference between the two groups (P> 0.05). CONCLUSION: Human esophageal smooth muscle cells express only Kir3.2, Kir3.3 and Kir3.4 in Kir3, and Kir3.4 is the most abundant in human esophageal smooth muscle cells. Expression of human LM and CM Kir3.2, Kir3.3 and Kir3.4 No significant difference.