摘要
目的基于环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/水通路蛋白(AQPs)通路以及胃肠紧密功能,观察江西龙虎山铁皮石斛不同提取工艺(水提、醇提、多糖)对胃阴虚小鼠模型干预作用机制。方法将雄性ICR小鼠120只随机分为正常组、模型组、益胃汤组(15 g/kg)、奥美拉唑组(1.7 mg/kg)、不同提取工艺铁皮石斛高剂量组(4.2 g/kg)、不同提取工艺铁皮石斛低剂量组(2.1 g/kg),每组12只。采用白酒和辣椒液的混合液灌胃建立胃阴虚小鼠模型,造模至第30天开始给药,持续至第70天,实验过程中小鼠每3天测定体重、饮水量、饮食量。实验结束后计算颌下腺指数、测定粪便含水量与唾液分泌量;HE染色观察胃组织变化;ELISA测定血清中血管活性肠多肽(VIP)、cAMP的含量;Western Blot检测cAMP/PKA/AQPs通路中PKA以及磷酸化PKA蛋白(p-PKA)、AQP3、AQP4、AQP5蛋白表达及紧密连接蛋白(ZO-1)、E-钙粘素(E-cadherin)表达。结果与正常组比较,模型组小鼠饮水量、体重、唾液分泌量、粪便含水率均下降(P<0.05),HE染色显示胃黏膜组织变薄、缺损,肌层丢失、腺体排列紊乱、炎性细胞浸润;血清中VIP、cAMP含量降低(P<0.05),蛋白p-PKA/PKA及AQP3、AQP4、ZO-1表达下调(P<0.05,P<0.01),AQP5、E-cadherin表达上调(P<0.05,P<0.01)。与模型组比较,给予不同铁皮石斛提取物后,水提高剂量、多糖高剂量组小鼠唾液分泌量升高(P<0.05),奥美拉唑及醇提、多糖组粪便含水率升高(P<0.05,P<0.01),3种提取物(除醇提低剂量组外)VIP、c AMP含量均升高(P<0.05),益胃汤组c AMP含量升高(P<0.05);3种提取物(除醇提高剂量组外)p-PKA/PKA比值上调(P<0.05,P<0.01),除益胃汤组外各组ZO-1表达均上调(P<0.05),3种提取物E-cadherin蛋白表达下调(P<0.05,P<0.01);奥美拉唑组及水提物、多糖高剂量组AQP3蛋白表达上调(P<0.05,P<0.01),3种提取物的高剂量AQP4蛋白表达均上调(P<0.01),多糖组AQP5蛋白表达下调(P<0.01,P<0.05)。�
Objective Through cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)/aquaporins(AQPs)pathway and gastrointestinal tract function,to investage the mechanism of Wei yin deficiency mice model based on the different extracting processes(water extract,alcohol extract and polysaccharide)of Dendrobium officinale from Longhu mountain in Jiangxi Province.Methods Totally 120 male ICR mice were randomly divided into normal group,model group,Yiwei Decoction group(15 g·kg^(-1)),Omeprazole group(1.7 mg·kg^(-1)),and also every different extracting processes including high dose of Dendrobium officinale group(4.2 g·kg^(-1)),low dose of Dendrobium officinale group(2.1 g·kg^(-1)),12 mice in each group.The Wei yin deficiency mice model was established by intragastric administration with a mixture of liquor and pepper liquid.The drug intervention groups received intervention starting from the 30th day of modeling and continued until the 70th day.The body weight,water and diet intake were measured every 3 days during the experiment.The submandibular gland index was calculated,and the fecal water content and saliva secretion were measured after the experiment.The gastric tissue was observed by HE staining.The contents of vasoactive intestinal peptide(VIP)and cyclic adenosine monophosphate(cAMP)in serum were determined by ELISA.The protein expressions of PKA,phosphorylated PKA,AQP3,AQP4,AQP5,Zonula occluden-1(ZO-1),and E-cadherin were determined by Western Blot.Results Compared with the normal group,the food intake,water intake,body weight,and the content of saliva secretion and fecal water decreased in the model group(P<0.05).HE staining showed that the gastric mucosal tissue became thinner and more defective,leading to the loss of the muscle layer and disordered,arrangement of glands,thus finally resulted in inflammatory cell infiltration.The content of VIP and cAMP in serum decreased(P<0.05),the expressions of p-PKA/PKA,AQP3,AQP4,and ZO-1 protein were down-regulated(P<0.05,P<0.01),the expressions of AQP5 and E-cadherin we
作者
赵益
卢雪凤
方红
吴柳瑾
陈丽华
朱培林
饶毅
ZHAO Yi;LU Xue-feng;FANG Hong;WU Liu-jin;CHEN Li-hua;ZHU Pei-lin;RAO Yi(Research Center of Differentiation and Development of Chinese Medicine Basic Theory,Jiangxi University of Chinese Medicine,Nanchang 330004;National Engineering Research Center for Manufacturing Technology of Solid Chinese Medicine Preparations,Jiangxi University of Chinese Medicine,Nanchang 330004;State Key Laboratory of Modern Preparation of Chinese Medicine,Ministry of Education,Jiangxi University of Chinese Medicine,Nanchang 330004;Jiangxi Academy of Forestry,Nanchang 330013)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2024年第9期1100-1108,共9页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然基金资助项目(No.81860777)
江西省重大科技研发专项(No.20203ABC28W019)
江西省科技计划“揭榜挂帅”关键技术类项目(No.20212AAF01005)
江西中医药大学校级科技创新团队发展计划(No.CXTD22007)
江西中医药大学中西医结合一级学科(江西省双一流学科)(No.zxyylxk20220103)。
关键词
铁皮石斛
铁皮石斛多糖
胃阴虚
水通道蛋白
紧密连接蛋白
中药
Dendrobium officinale
Dendrobium officinale polysaccharides
Wei yin deficiency
aquaporin
zonula occludes protein
Chinese herbal medicine
