【摘要】 目的：探讨麻杏配伍类方（麻黄汤、三拗汤、麻杏石甘汤）在小鼠哮喘模型中对温度敏感瞬时受体电位（TRP）离子通道调控作用的差异性。方法：雌性C57BL/6小鼠60只，分为空白组、模型组、地塞米松组(0.75 mg·kg^-1)、麻黄汤组(4.1 g·kg^-1)、三拗汤组(2.7 g·kg^-1)、麻杏石甘汤组(6.4 g·kg^-1)，每组10只。采用卵清蛋白（OVA）复制哮喘小鼠模型，模型建立后第19～28天，各给药组灌胃给药(10 mL·kg^-1)，空白组予生理盐水，每天1次。观察各组小鼠外周血嗜酸性粒细胞（EOS）和支气管肺泡灌洗液（BALF）中白细胞（WBC）水平、呼吸间歇（Penh）值变化、肺组织病理损伤，蛋白免疫印迹法（Western blot）和实时荧光定量聚合酶链式反应（Real-time PCR）检测小鼠肺组织高温敏感通道TRPV1、TRPV3和低温敏感通道TRPA1、TRPM8蛋白与mRNA表达。结果：与空白组比较，模型组小鼠表现出典型的哮喘表型，外周血EOS和BALF中WBC水平、Penh值明显升高（P<0.05,P<0.01），肺组织损伤严重。与模型组比较，三方干预后均可不同程度减轻模型小鼠的哮喘表型（P<0.05,P<0.01）。与空白组比较，模型组小鼠肺组织TRPV1、TRPA1蛋白表达显著上调（P<0.01）；与模型组比较，麻杏石甘汤、三拗汤组TRPV1、TRPA1蛋白表达明显下调（P<0.05,P<0.01），且两方组间比较，麻杏石甘汤组TRPV1蛋白表达降低更显著（P<0.01），三拗汤组TRPA1蛋白表达降低更显著（P<0.01），麻黄汤组未见明显调控作用。进一步检测结果表明，与空白组比较，模型组TRPV3、TRPM8蛋白表达显著上调（P<0.01）；与模型组比较，麻杏石甘汤、三拗汤组TRPV3、TRPM8蛋白表达显著下调（P<0.01），两方组间比较，麻杏石甘汤组TRPV3蛋白表达降低更明显（P<0.05），三拗汤组TRPM8蛋白表达降低更显著（P<0.01）。与空白组比较，模型组小鼠肺组织TRPV1、TRPV3、TRPA1、TRPM8 mRNA水平显著升高（P<0.01），三拗汤、麻杏石甘汤显著降低模型小鼠肺组织中TRPV1、TRPV3、TRPA1、TRPM8 mRNA水平（P<0.01），麻杏石甘汤组对高温敏感通道蛋白TRPV1、TRPV3的调控作用明显优于三拗汤组（P<0.05,P<0.01），三拗汤组对低温敏感通道蛋白TRPA1、TRPM8的调控作用明显优于麻杏石甘汤组（P<0.05,P<0.01）。结论：麻黄汤、三拗汤、麻杏石甘汤均可减轻哮喘小鼠哮喘气道炎症，麻黄汤未表现出对TRPV1、TRPA1的调控作用，麻杏石甘汤偏于调节高温敏感TRPV1、TRPV3通道，三拗汤偏于调节低温敏感TRPA1、TRPM8通道。更多还原

【Abstract】 Objective: To investigate the differences in the regulatory effects of formulas containing Ephedrae Herba and Armeniacae Semen Amarum（Mahuangtang, Sanaotang, and Maxing Shigantang） on thermosensitive transient receptor potential ion channels（thermo TRPs） in the mouse model of asthmatic airway inflammation. Methods: Sixty female C57 BL/6 mice were allocated into blank, model, dexamethasone(0.75 mg·kg^-1), Mahuangtang(3.8 g·kg^-1), Sanaotang(2.8 g·kg^-1), and Maxing Shigantang(6.6 g·kg^-1) groups（n=10）. The mouse model of asthma was established with ovalbumin（OVA） and treated with normal saline（blank group） or corresponding drugs(10 mL·kg^-1), once a day, 19-28 days after modeling. The levels of eosinophils（EOS） in peripheral blood and white blood cell（WBC） in bronchoalveolar lavage fluid（BALF）, changes in enhanced pause（Penh）, and pathological damage of lung tissue were observed in each group. Western blot and real-time PCR were employed to quantify the protein and mRNA levels, respectively, of high-temperature thermosensitive channels（TRPV1 and TRPV3） and low-temperature thermosensitive channels（TRPA1 and TRPM8） in the lung tissue. Results: Compared with the blank group, the model group showed a typical asthma phenotype, including elevations in the level of EOS in peripheral blood, level of WBC in BALF, and value of Penh（P<0.05,P<0.01）, and severe lung tissue damage. Compared with the model group, the three formulas alleviated the asthma phenotype to varying degrees（P<0.05,P<0.01）. Compared with the blank group, the model group showed upregulated protein levels of TRPV1 and TRPA1 in the lung tissue（P<0.01）. Compared with the model group, Maxing Shigantang and Sanaotang groups showed down-regulated protein levels of TRPV1 and TRPA1（P<0.05, P<0.01）. Moreover, Maxing Shigantang and Sanaotang groups showed more significant down-regulation in protein levels of TRPV1 and TRPA1, respectively（P<0.01）, while no obvious regulatory effect was observed in the Mahuangtang group. Compared with those in the blank group, the protein levels of TRPV3 and TRPM8 were up-regulated in the model group（P<0.01）. Compared with the model group, Maxing Shigantang and Sanaotang down-regulated the protein levels of TRPV3 and TRPM8（P<0.01）. Moreover, Maxing Shigantang and Sanaotang exerted stronger down-regulating effects on TRPV3（P<0.05） and TRPM8（P<0.01）, respectively. Compared with the blank group, the model group presented up-regulated mRNA levels of TRPV1, TRPV3, TRPA1, and TRPM8 in the lung tissue（P<0.01）, and such up-regulations were significantly decreased by Maxing Shigantang and Sanaotang（P<0.01）. Moreover, Maxing Shigantang outperformed Sanaotang in regulating high-temperature thermosensitive channels TRPV1 and TRPV3（P<0.05, P<0.01）. The regulation effect of the, Maxing Shigantang on high-temperature thermosensory channel proteins of TRPV1 and TRPV3 was better than that of the Sanaotang P<0.05 P<0.01 while the Sanaotang outperformedhad a significant regulatory effect on Maxing Shigantang in regulating the low-temperature thermosensory thermosensitive channel proteins of TRPA1 and TRPM8 which was better than that of the Maxing Shigantang（P<0.05,P<0.01）. Conclusion: The experimental results showed that Mahuangtang, Sanaotang, and Maxing Shigantang all had protective effects on asthma airway inflammation. while Mahuangtang did not show the regulatory effect on TRPV1 and or TRPA1. Maxing Shigantang preferred to regulate high-temperature thermosensory thermosensitive channels of TRPV1 and TRPV3 channels, and Sanaotang preferred to regulate low-temperature thermosensory thermosensitive channels of TRPA1 and TRPM8.更多还原