基于PKA依赖cAMP信号介导肺胃SP表达探讨中医肺胃相关理论的机制研究  

Role of protein kinase A(PKA)-dependent cyclic adenosine monophosphate(cAMP)signaling-mediated substance P(SP)expression in lung,gastric,and esophageal tissues in a rat model of the gastroesophageal reflux,further confirmation of TCM lung/stomach-rela

作　　者：晏露宁 汝触会 陈爱凤[1] 陈晔 韩佳颖 何飞 YAN Lu-ning;RU Chu-hui;CHEN Ai-feng;CHEN Ye;HAN Jia-ying;HE Fei(Department of Pulmonary and Critical Care,Hangzhou Chest Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China;Department of Pulmonary and Critical Care,The Second Affiliated Hospital to Zhejiang University of Traditional Chinese Medicine,Hangzhou 310005,China)

机构地区：[1]浙江大学医学院附属杭州市胸科医院呼吸与危重症医学科,杭州310003 [2]浙江中医药大学附属第二医院呼吸与危重症医学科,杭州310005

出　　处：《浙江中西医结合杂志》2022年第11期997-1000,1007,共5页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.2019ZA088)

摘　　要：目的观察胃食管反流模型大鼠肺、胃、食管组织中蛋白激酶A(PKA)、环磷酸腺苷(cAMP)、P物质(SP)的表达量,探讨中医肺胃相关理论的分子机制。方法将12只SD大鼠随机数字表法分为正常对照组(n=6)和模型组(n=6),模型组大鼠麻醉后经口插胃管至食管的中下段,以8滴/min的速率缓慢灌注0.1 mmol/L盐酸(含0.5%胃蛋白酶)溶液,每次20 min,1次/d,连续14 d。正常对照组采用PBS液代替,方法同模型组。提取食管、胃和肺组织标本进行病理学观察,用免疫组化法测定PKA、cAMP、SP表达,并将结果进行线性回归分析。结果与正常对照组比较,模型组大鼠肺、胃及食管组织HE半定量评分显著升高[(3.67±0.52)分比(0.00±0.00)分、(3.83±0.41)分比(0.00±0.00)分、(3.83±0.41)分比(0.00±0.00)分,P均<0.01]。与正常对照组比较,模型组大鼠肺、食管、胃组织的PKA蛋白表达量[(0.23±0.02)比(0.15±0.01)、(0.22±0.02)比(0.17±0.02)、(0.24±0.02)比(0.16±0.01),P均<0.01]、cAMP蛋白表达量[(0.20±0.02)比(0.15±0.02)、(0.24±0.03)比(0.15±0.02)、(0.25±0.03)比(0.17±0.02),P均<0.01]、SP蛋白表达量[(0.21±0.02)比(0.15±0.01)、(0.25±0.03)比(0.14±0.02)、(0.33±0.04)比(0.18±0.02),P均<0.01]均显著升高。线性回归结果表明,模型组大鼠胃与肺组织的PKA和SP平均光密度呈线性相关。结论激活后的PKA/cAMP是促进肺胃组织释放SP的重要通路,可能介导了胃食管反流咳嗽的病理生理过程,是肺胃相关理论可能的分子机制之一。Objective To assess of the TCM lung/stomach related theory based on the protein kinase A(PKA)-dependent cyclic adenosine monophosphate(cAMP)signal-mediated substance P(SP)expression in lung,gastric,and esophageal tissues in a rat model of the gastroesophageal reflux.Methods Twelve Sprague-Dawley rats were randomly divided into normal control and model groups(n=6 per group).The model rats were anesthetized and inserted the gastric tube into the middle and lower part of the esophagus and 0.1 mmol/L hydrochloric acid containing 0.5%pepsin solution was slowly perfused into the tube at a rate of eight drops 20 min daily for 14 consecutive days,while the control rats were given saline instead.After that,esophageal,gastric,and lung tissue samples were resected from rats for histology and expressions of PKA,cAMP,and SP using H&E staining and immunohistochemistry,respectively.The results were analyzed by linear regression.Results Compared with those of the control rats,there were significant damages in the lung,gastric and esophageal tissues of the model rats were significantly damages in terms of histology scores(3.67±0.52,3.83±0.41,and 3.83±0.41 vs.0.00±0.00 points,respectively;P<0.01).Compared with the control group,expression of PKA(0.23±0.02 vs.0.15±0.01,0.22±0.02 vs.0.17±0.02,and0.24±0.02 vs.0.16±0.01,respectively;P<0.01),cAMP(0.20±0.02 vs.0.15±0.02,0.24±0.03 vs.0.15±0.02,and0.25±0.03 vs.0.17±0.02,respectively;P<0.01),and SP proteins(0.21±0.02 vs.0.15±0.01,0.25±0.03 vs.0.14±0.02,and 0.33±0.04 vs.0.18±0.02,respectively;P<0.01)were significantly increased in the lung,esophageal and gastric tissues in the model rats.The linear regression results showed that the average optical density of PKA and SP in the gastric and lung tissues in the model rats was linearly correlated.Conclusion The activated PKA/cAMP is an important pathway to promote the release of SP protein in the lung and gastric tissues,which may mediate the pathophysiological process of gastroesophageal reflux cough,one of the possible m

关 键 词：P物质 蛋白激酶A 环磷酸腺苷(cAMP) 肺胃相关 

分 类 号：R256[医药卫生—中医内科学]