舒胃汤对功能性消化不良肝郁脾虚证模型大鼠胃窦组织中ENS-PDGFRα^(+)细胞-SMC网络超微结构的影响  

Exploring the Effect of Shuwei Decoction(舒胃汤)for Rats with Functional Dyspepsia of Liver-Depression and Spleen-Deficiency Syndrome Model on the Ultrastructure of ENS-PDGFRα^(+)Cell-SMC Network in Gastric Sinusoidal Tissues

作　　者：梁俊尧 郭璇[1] 张德旭 周曼 肖婕 唐楚森 封慧 徐寅[1] LIANG Junyao;GUO Xuan;ZHANG Dexu;ZHOU Man;XIAO Jie;Tang Chusen;Feng Hui;XU Yin(The First Hospital of Hunan University of Chinese Medicine,Changsha,410000)

机构地区：[1]湖南中医药大学第一附属医院,湖南省长沙市410000

出　　处：《中医杂志》2024年第12期1269-1278,共10页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81904176);湖南省自然科学基金(2023JJ60044);湖南省研究生科研创新项目(CX20230834);湖南省教育厅科学研究项目(21B0389);湖南省教育厅重点课题(19A375);湖南省中医药科研课题(B2023079);湖南省科技创新计划项目(2021SK51413);湖南中医药大学中医学“十三五”一级学科建设项目。

摘　　要：目的基于肠神经系统(ENS)-血小板衍生的生长因子受体α阳性(PDGFRα^(+))细胞-平滑肌细胞(SMC)网络超微结构探讨舒胃汤治疗功能性消化不良(FD)肝郁脾虚证的可能作用机制。方法将50只SD大鼠随机分为空白组、模型组及舒胃汤低、中、高剂量组,每组10只。空白组不予造模,其余大鼠采用改良复合病因造模法(慢性束缚应激+夹尾激怒+过度疲劳+饮食失节)建立FD肝郁脾虚证大鼠模型,持续21天。造模完成后舒胃汤低、中、高剂量组大鼠分别给予舒胃汤药液3.78、7.56、15.12 g/(kg·d)灌胃,空白组和模型组大鼠给予生理盐水10 ml/(kg·d)灌胃,均每日灌胃1次,持续14天。实验结束后比较各组大鼠灌胃前后体质量增长值,进行高架十字迷宫实验观察大鼠开臂停留时间百分比和进入开臂次数百分比;黑色半固体糊灌胃法检测大鼠胃排空率、小肠推进率;HE染色法观察胃窦组织病理情况;Western Blot法和RT-qPCR法分别检测胃窦组织血小板衍生的生长因子受体α(PDGFRα)、小电导钙激活钾通道3(Sk3)、嘌呤能P2Y1受体(P2Y1)蛋白及mRNA表达;免疫荧光双染检测胃窦组织中PDGFRα与酪氨酸激酶受体(C-kit)、蛋白基因产物9.5(PGP9.5)、Sk3、平滑肌细胞(SMC)的共定位情况;透射电镜观察ENS-PDGFRα^(+)细胞-SMC网络的超微结构。结果HE染色结果显示,各组大鼠胃组织黏膜皱襞清晰,未见出血、溃疡、肿胀等器质性病变,模型组及舒胃汤低剂量组胃窦组织固有层内部分区域胃腺排列较为紊乱,舒胃汤中、高剂量组固有层内胃腺排列较为整齐,病变程度轻于模型组及舒胃汤低剂量组。与空白组比较,模型组大鼠灌胃前后体质量增长值、开臂停留时间百分比、进入开臂次数百分比、小肠推进率、胃排空率均明显降低,胃窦组织中PDGFRα、Sk3、P2Y1蛋白及mRNA表达降低,PDGFRα与C-kit、Sk3、SMC、PGP9.5共定位表达明显降低(P<0.05或P<0.01),ENS-Objective Based on the ultrastructure of enteric nervous system(ENS)-platelet-derived growth factor receptorαpositive(PDGFRα^(+))cell-smooth muscle cell(SMC)network,the effect and mechanism of Shuwei Decoction(舒胃汤)for rats with functional dyspepsia(FD)of liver-depression and spleen-deficiency syndrome were explored.Methods Fifty SD rats were divided into blank group,model group,low-,medium-and high-dose Shuwei Decoction groups randomly,with 10 rats in each group.The blank group was not set up,and the other 4 groups were set up with the modified composite cause(chronic restraint stress+tail irritation+excessive fatigue+diet disorder)for 21 days,resulting in FD model rats with liver-depression and spleen-deficiency syndrome.After successful modelling,rats in the low-,medium-and high-dose groups were given 3.78,7.56 and 15.12 g/(kg·d)of Shuwei Decoction by gavage,respectively,while rats in the blank group and the model group were given 10 ml/(kg·d)of saline by gavage;all of them were given gavage once a day for 14 days.Body mass increase were collected before and after gavage,and compared after the experiment;the elevated cross maze experiment was conducted to observe the percentage of staying time and the percentage of times entering the open arm of rats;the gastric emptying rate and small intestinal propulsion rate were measured by black semi-solid paste;HE staining was used to observe histopathology of the gastric antrum;the Western Blotting and RT-qPCR were used to detect the expression levels of PDGFRα,small conductance calcium activated potassium(SK3),and purinergic G protein-coupled receptor P2Y1(P2Y1 R),and mRNA expression in gastric antrum;immunofluorescence double staining was used to detect the co-localization of PDGFRαwith receptor tyrosine kinase(C-kit),protein gene product 9.5(PGP9.5),Sk3,and smooth muscle cells(SMCs)in gastric sinusoidal tissues;and transmission electron microscopy was used to observe the ultrastructure of the ENS-PDGFRα^(+)cell-SMC network.Results HE staining results sho

关 键 词：功能性消化不良 肝郁脾虚证 舒胃汤 胃肠动力 肠神经系统 血小板衍生的生长因子受体α 平滑肌细胞 超微结构 

分 类 号：R285.5[医药卫生—中药学]