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目的:观察不同压力对家犬气道黏液层黏蛋白(MUC)分泌的影响,并初步探讨其参与机制。方法:24只健康家犬行双腔支气管导管插管,并随机分为4组(n=6)。A组家犬正常呼吸的频率及压力双侧通气(A1,A2);B组一侧不通气(B1),另一侧予以过量通气(B2);C组预先张力敏感性阳离子通道4(TRPV4)阻断剂钉红(RR)后按A组通气模式通气(C1,C2);D组预先TRPV4阻断剂RR处理后按B组通气模式通气(D1,D2)。通气12 h后,ELISA检测3组支气管灌洗液(BALF)中MUC(2,5AC和5B)蛋白含量;RT-PCR检测支气管肺组织匀浆中MUC(2,5AC和5B)mRNA转录水平。结果:与正常通气的A2组比较,过量通气的B2组家犬BALF中3种MUC蛋白含量显著升高,且以MUC5AC为主(P<0.05);与A1组比较,未通气的B1组则明显下降(P<0.05)。与A1,A2组比较,给予RR处理后的C1,C2组蛋白含量均明显降低(P<0.05)。与过量通气的B2组比较,给予TRPV4特异性阻断剂处理后再过量通气的D2组3种MUC蛋白含量亦明显下降(P<0.05)。与A2组比较过量通气的B2组肺组织MUC(2,5AC和5B)mRNA表达均明显上调(P<0.05);与A1,A2比较,给予RR处理后C1,C2组肺组织3种MUC mRNA转录水平显著下降(P<0.05);给予RR预处理后与B2组相比,D2组内MUC mRNA转录水平显著下调(P<0.05)。结论:节律性压力波参与维持家犬气道上皮黏液层黏蛋白分泌平衡调控,其机制主要通过TRPV4通道实现。
Objective: To observe the effects of different pressures on mucus laminin (MUC) secretion in the airway of dogs and to explore its mechanism. Methods: Twenty-four healthy domestic dogs underwent double lumen bronchial catheterization and were randomly divided into 4 groups (n = 6). A group of dogs normal breathing frequency and pressure of bilateral ventilation (A1, A2); B group on the one side without ventilation (B1), the other side to be over-ventilation (B2); C group pre-tension sensitive cation channel 4 (TRPV4 ) Blocking agent nail red (RR) ventilation group A ventilation mode (C1, C2); Group D pre-TRPV4 blocker RR treatment group B ventilation mode ventilation (D1, D2). The contents of MUC (2,5AC and 5B) in bronchoalveolar lavage fluid (BALF) were detected by ELISA 12 h after ventilation. The mRNA transcription levels of MUC (2,5AC and 5B) in bronchoalveolar homogenate were detected by RT-PCR. Results: Compared with A2 group, the content of three MUC proteins in BALF of over-aeration group B2 was significantly increased (P <0.05). Compared with group A1, the non-ventilated group B1 Decreased significantly (P <0.05). Compared with group A1 and group A2, the contents of C1 and C2 protein in RR group were significantly decreased (P <0.05). Compared with over-aerated group B2, the contents of three MUC proteins in D2 group that were over-aerated with TRPV4-specific blockers were also significantly decreased (P <0.05). The mRNA expression of MUC (2,5AC and 5B) in group B2 was significantly increased compared with group A2 (P <0.05). Compared with group A1 and group A2, MUC mRNA expression in lungs of group C1 and C2 (P <0.05). After pretreatment with RR, the transcription level of MUC mRNA in D2 group was significantly lower than that in B2 group (P <0.05). CONCLUSION: Rhythmic pressure wave is involved in the regulation of the balance of mucin secretion in the airway epithelial mucosa in dogs. The mechanism is mainly through TRPV4 channel.