目的 探讨健脾清化颗粒对胃食管反流病（Gastroesophageal reflux disease，GERD）脾虚湿热证大鼠食管黏膜肥大细胞（Mast cells， MCs）增殖活化的调节作用，及对类胰蛋白酶（mast cell tryptase，MCT）、蛋白酶激活受体-2（protease activated receptors-2，PAR2），瞬时受体电位香草酸受体1（transient receptor potential vanilloid1，TRPV1）表达的影响。方法 采用改良食管十二指肠侧侧吻合术叠加内外因湿热干预法建立GERD脾虚湿热证病证结合大鼠模型。将72只SD大鼠随机分为假手术组（12只）和模型组（60只），模型评价后将大鼠随机分为5组，分别是模型组、奥美拉唑组、健脾清化颗粒低剂量组、中剂量组、高剂量组，每组12只。各给药组连续灌胃14天，假手术组、模型组给予蒸馏水灌胃。采用HE染色观察大鼠食管黏膜的形态学改变；甲苯胺蓝染色观察大鼠食管黏膜MCs的形态和数量；免疫组化检测大鼠食管黏膜MCT、PAR2、TRPV1的表达情况。结果 与假手术组比，模型组大鼠体重、进食量下降（P<0.05），肛温、粪便含水量升高（P<0.01）；食管黏膜基底细胞增生，炎性细胞浸润，乳头肌延长；MCs数量增多（P<0.01）；免疫组化MCT、PAR2、TRPV1平均光密度值（mean optical density，MOD）增加（P<0.05）。与模型组比，健脾清化颗粒各组给药前后大鼠体重、进食量增加，肛温、粪便含水量下降（P<0.05或P<0.01），健脾清化颗粒各组食管黏膜MCs数量减少（P<0.01）；MCT、PAR2、TRPV1的表达减少（P<0.05或P<0.01）。结论 GERD脾虚湿热证模型大鼠食管黏膜MCs增殖活化，MCT、PAR2、TRPV1的表达增加。健脾清化颗粒能够改善大鼠食管黏膜MCs的增殖活化，下调MCT、PAR2、TRPV1的表达，对改善食管高敏感机制发挥一定的作用。

Objective To investigate the effect of Jianpi Qinghua Granule on the proliferation and activation of mast cells (MCs) and the expression of tryptase (MCT), protease activated receptor-2 (PAR2) and transient receptor potential vanillic acid receptor 1 (TRPV1) in rats with gastroesophageal reflux disease (GERD) of spleen deficiency and damp heat.Methods 72 SD rats were randomly divided into sham operation group (n=12) and model group (n=60). The model group was established by esophagoduodenostomy (EGDA) combined with intervention of damp-heat syndrome. After model evaluation, the rats were randomly divided into model group, low-dose group of Jianpi Qinghua granule, medium-dose group of Jianpi Qinghua granule, high-dose group of Jianpi Qinghua granule and omeprazole group.12 rats in each group were given intragastric administration for 14 days, while in sham group, distilled water was given intragastric administration. HE staining was used to observe the morphological changes of esophageal mucosa, toluidine blue staining was used to observe the activation of MCs, and immunohistochemistry was used to detect the expression of MCT, PAR2 and TRPV1 protein.Results Compared to the sham group, the body weight and food intake of the model group decreased(P<0.05), the anal temperature and fecal water content increased in the model group (P<0.01). The esophageal mucosa in the model group :basal cell prolifeation, papillary muscle prolongation,inflammatory cell infiltration were more obviouse.,,the number of MCs in the model group was increased (P<0.01). ,the MOD of MCT,,PAR2 and TRPV1 were increased (P<0.05). Compared to the model group, the body weight and food intake increased the anal temperature and fecal water content decreased after adminisitration in each JPQH group(P<0.05or P<0.01),the number of MCs in the each JPQH group was decreased (P<0.01) , the MOD of MCT, PAR2 and TRPV1 in each JPQH group was decreased .(P<0.05或P<0.01).Conclusion the expression of MCT, PAR2 and TRPV1 in the esophageal mucosa rats with GERD of spleen deficiency and damp-heat syndrome is increased. Jianpi Qinghua granule produce an effect of inhibiting the proliferation and activation of MCs in rat esophageal mucosa, reducing the expression of MCT, PAR2 and TRPV1, which play a certain role in alleviating esophageal hypersensitivity.

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国家科学技术部国家重点研发计划（SQ2019YFC170077）:基于脾胃通降理论系列方辨证治疗非糜烂性反流病的疗效优势和机制研究；负责人：唐旭东。中国中医科学院西苑医院苗圃课题（2019XYMP-48）:肥大细胞活化PAR2/TRPV1通路在NERD食管高敏感中的外周机制及健脾清化颗粒的干预研究；负责人：车慧。