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目的研究大鼠急性肺栓塞模型肺组织中与酸碱平衡相关的碳酸酐酶3(CA3)和Na+/H+交换器(NHE1)的表达变化。方法建立大鼠急性肺栓塞模型,分别在急性肺栓塞后1、8、24和48 h开胸取出肺组织,然后提取肺组织的总RNA和总蛋白,以正常组为对照,采取半定量RT-PCR的方法研究CA3和NHE1在mRNA水平表达的变化;采用western-b lot方法进一步验证CA3和NHE1在蛋白水平表达的变化;同时采用免疫组织化学的方法检测大鼠肺组织中NHE1在肺栓塞前后表达的变化。结果在大鼠急性肺栓塞后的不同时间点,CA3的mRNA水平和蛋白水平均逐渐降低,而NHE1在mRNA水平和蛋白水平的表达都出现逐渐升高现象。免疫组化研究表明NHE1主要分布在支气管黏膜上皮和细支气管黏膜上皮的胞浆内,急性肺栓塞后NHE1在上皮细胞内的表达明显升高。结论大鼠急性肺栓塞后肺组织内CA3的表达降低,NHE1的表达升高,这一现象可能与机体的酸碱平衡调节相关。
Objective To investigate the expression changes of carbonic anhydrase 3 (CA3) and Na + / H + exchanger (NHE1) in the lung tissue of rats with acute pulmonary embolism. Methods Acute pulmonary embolism model was established in rats. Pulmonary tissues were obtained by thoracotomy at 1, 8, 24 and 48 h after acute pulmonary embolism. Total RNA and total protein in lung tissue were extracted. The normal group was used as a control and semi-quantitative RT -PCR was used to study the expression of CA3 and NHE1 at mRNA level. Western-b lot method was used to further verify the expression of CA3 and NHE1 at protein level. Immunohistochemistry was used to detect the expression of NHE1 in pulmonary embolism Changes before and after the expression. Results The mRNA and protein levels of CA3 decreased gradually at different time points after acute pulmonary embolism in rats. However, the expressions of NHE1 at mRNA and protein levels gradually increased. Immunohistochemical studies showed that NHE1 mainly distributed in the cytoplasm of bronchial mucosa epithelium and bronchiolar mucosal epithelium, NHE1 expression in epithelial cells was significantly increased after acute pulmonary embolism. Conclusions The expression of CA3 and the expression of NHE1 in the lungs of rats after acute pulmonary embolism are decreased, which may be related to the regulation of acid-base balance.