蝉芩颗粒通过下调COX-2抑制TRPV1通道减轻感染后咳嗽大鼠气道神经源性炎症

Chanqin Granules Alleviates Airway Neurogenic Inflammation of Post-Infectious Cough Through Down-regulating COX-2 and Inhibiting TRPV1 Channel

【目的】观察蝉芩颗粒不同剂量对感染后咳嗽大鼠气道神经源性炎症的作用机制。【方法】将SD大鼠随机分为空白组,模型组,蝉芩颗粒低、中、高剂量组及阿斯美组。除空白组,其他各组采用烟熏结合内毒素滴鼻及辣椒素刺激复制感染后咳嗽模型。造模成功后,各给药组对应灌胃给药,模型组给予等体积生理盐水,每天1次,连续14 d,空白组不做任何处理。以酶联免疫吸附分析(ELISA)检测血清中P物质(SP)含量;逆转录聚合酶链反应(RT-PCR)法检测肺组织和背根神经节中环氧化酶2(COX-2)、前列腺素E2受体亚型3(EP3)、瞬时受体电位类香草酸受体亚型1(TRPV1) m RNA水平;蛋白免疫印迹(Western Blot)法检测TRPV1通道蛋白表达的影响。【结果】(1)ELISA结果:蝉芩颗粒各剂量组、阿斯美组血清中SP含量较模型组下降(P <0.05)。(2)RT-PCR结果:蝉芩颗粒高剂量组肺组织和背根神经节中COX-2 mRNA表达水平低于模型组(P <0.05);蝉芩颗粒各剂量组、阿斯美组肺组织和背根神经节中EP3 m RNA表达水平低于模型组,但差异无统计学意义(P> 0.05);蝉芩颗粒各剂量组、阿斯美组肺组织中TRPV1 m RNA表达水平低于模型组(P <0.01),蝉芩颗粒各剂量组背根神经节中TRPV1 mRNA表达水平低于模型组(P <0.05或P <0.01)。(3)Western Blot结果:蝉芩颗粒各剂量组、阿斯美组肺组织和背根神经节中TRPV1蛋白表达水平低于模型组(P <0.01)。【结论】蝉芩颗粒可减轻感染后咳嗽大鼠气道神经源性炎症,其机制可能与下调COX-2及TRPV1表达、减少速激肽SP的释放有关。

Objective To explore the action mechanism of Chanqin Granules for airway neurogenic inflammation of rats with post-infectious cough. Methods The SD rats were randomly divided into blank group,model group,low-,middle-and high-dose Chanqin Granules groups and Asme group. Apart from the blank group,the postinfectious cough rat model was established by fumigation combined with lipopolysaccharide nasal drops and capsaicin stimulation. After modeling, the medication groups were given intragastric administration of corresponding drug,the model group was given intragastric administration of the same volume of normal saline,one time per day,the treatment lasting 14 continuous days,additionally,the blank group had no treatment. The serum content of substance P(SP)was detected by enzyme-linked immunosorbent assay(ELISA),the mRNA levels of cyclooxygenase 2(COX-2),prostaglandin E2 receptor subtype 3(EP3)and transient receptor potential vanilloid 1(TRPV1)in lung tissue and dorsal root ganglion were measured by reverse transcription polymerase chain reaction(RT-PCR),and the expression level of TRPV1 protein was measured by Western blotting method.Results(1)ELISA results showed that the serum SP content in various Chanqin Granule groups and Asme group was decreased as compared with that in the model group(P < 0.05).(2)RT-PCR results showed that the mRNA expression level of COX-2 in lung tissue and dorsal root ganglion of high-dose Chanqin Granules group was decreased as comparion with that of the model group(P < 0.05);the mRNA expression level of EP3 in lung tissue and dorsal root ganglion of various Chanqin Granules groups and Asme group was lower than that of the model group,but there was no significant difference(P > 0.05);the m RNA expression level of TRPV1 in lung tissue of various Chanqin Granules groups and Asme group was lower than that of the model group(P < 0.01);the mRNA expression level of TRPV1 in dorsal root ganglion of various Chanqin Granules groups was lower than that of the model group(P < 0.05 or P < 0.01).(3)Western blotting results showed that TRPV1 protein expression level in lung tissue and dorsal root ganglion of various Chanqin Granules and Asme group was decreased as comparion with that of the model group(P < 0.01). Conclusion Chanqin Granules has an effect on alleviating airway neurogenic inflammation,and its mechanism is related with regulating the expression of COX-2 and TRPV1 and reducing the release of SP.