麸炒苍术对脾虚证大鼠结肠组织通透性的影响  

Effect of bran-fried Rhizoma Atractylodis on colonic permeability in rats with spleen deficiency syndrome

作　　者：谢慧臣 冉云 张云 欧阳瑜 唐浪 吴广阳 XIE Huichen;RAN Yun;ZHANG Yun;OUYANG Yu;TANG Lang;WU Guangyang(School of Traditional Chinese Medicine,Hubei Minzu University,Enshi 445000,Hubei,China;Department of Chinese Medicine,Enshi De Yuan Sheng Chinese Medicine Hospital,Enshi 445000,Hubei,China)

机构地区：[1]湖北民族大学中医药学院,湖北恩施445000 [2]恩施德元升中医医院中医科,湖北恩施445000

出　　处：《暨南大学学报（自然科学与医学版）》2023年第2期137-146,共10页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：湖北省自然科学基金知识创新专项项目(2019CFB781)。

摘　　要：目的:研究麸炒苍术对脾虚证大鼠结肠组织通透性的影响,探讨麸炒苍术改善脾虚证的可能机制。方法:60只雄性SD大鼠随机分为正常组(CON)、模型组(MOD)、麸炒苍术高、中、低剂量组(BRA-H、BRA-M、BRA-L)、复方谷氨酰胺组(CG),10只/组。除CON组外的5组均以慢性束缚、苦寒破气加饥饱失常法复制脾虚证模型,持续21 d。造模完成后,BRA-L、BRA-M、BRA-H组每kg大鼠体质量每天分别按5、10、20 g配制的中药煎剂2 mL灌胃,CG组按9 mg配制的复方溶液2 mL灌胃,余两组灌胃等体积的0.9%NaCl溶液,1次/d,持续14 d。后观察各组大鼠结肠组织病理学改变;使用ELISA法检测结肠黏膜组织ZO-1、Claudin-1、Claudin-2和Occludin的质量分数;使用免疫荧光法检测结肠黏膜组织细胞中p-MEK1/2及p-ERK1的蛋白表达;使用免疫组化与实时荧光定量PCR法检测各组大鼠结肠组织细胞中MLCK、CAM蛋白和基因的表达。结果:与MOD组比较,CON、BRA-L、BRA-M、BRA-H及CG组大鼠结肠黏膜组织ZO-1、Claudin-1和Occludin表达水平显著更高,Claudin-2表达水平显著更低(P<0.05,P<0.01);光镜及透射电镜下,MOD组结肠黏膜上皮及固有膜内基本结构被破坏,各治疗组则不同程度修复;MOD组胞内线粒体数量明显下降且结构明显异常,各治疗组则不同程度改善;与MOD组比较,CON、BRA-L、BRA-M、BRA-H及CG组结肠组织细胞中p-MEK1/2及p-ERK1蛋白表达、MLCK蛋白和基因表达显著更低,CAM蛋白和基因表达显著更高(P<0.05,P<0.01)。结论:麸炒苍术可通过调节结肠组织通透性及CAM-MLCK信号通路,双相调节结肠黏膜Claudin-1、Claudin-2等具有不同功能的紧密连接蛋白,增强肠上皮细胞屏障,修复受损的结肠黏膜组织结构,保护胃肠功能,较好地调节和改善脾虚证大鼠结肠病理结构和功能。Objective:To explore the effect of bran-fried Rhizoma Atractylodis on the colonic permeability of spleen deficiency rats,and clarify the possible mechanisms of bran-fried Rhizoma Atractylodis in improving spleen deficiency syndrome.Methods:Sixty male SD rats were randomly divided into the normal control group(CON),model group of(MOD),bran-fried Rhizoma Atractylodis groups of high,medium and low dose(BRA-H,BRA-M,BRA-L),compound glutamine group(CG),and 10 in each group.All the five groups except the CON group were treated with chronic restraint,bitter cold and broken qi combined with hunger and satiety disorder to replicate spleen deficiency model,and the modeling time was 21 days.After modeling,rats in BRA-H,BRA-M,BRA-L groups were treated with 5,10,20 g of Chinese herbal decoction per kg body weigh per day in 2 mL,and the CG group with 9 mg.The other two groups were samely administered with 0.9%NaCl solution,once a day for 14 days.After treatment,the histopathological changes of colon were observed in each group,and the content of ZO-1,claudin-1,claudin-2 and Occludin in colon mucosal tissues was detected by ELISA,and the protein expressions of P-MEK1/2 and P-ERK1 in colon mucosal tissues were detected by immunofluorescence assay.The expression of MLCK and CAM protein and gene in colon tissues were detected by immunohistochemistry and real-time quantitative PCR.Results:Compared with MOD group,the expression levels of ZO-1,claudin-1 and occludin in colonic mucosal tissue of the CON,BRA-L,BRA-M,BRA-H and CG group were significantly increased,and the expression of claudin-2 was significantly decreased(P<0.05,P<0.01).Under optical microscope and transmission electronic microscope,the basic structure of colonic mucosa epithelium and lamina propria in MOD group rats was destroyed,and the damaged structure of colonic mucosa epithelium and lamina propria of each treatment group was restored to a different degree.The number of intracellular mitochondria reduced markedly and obviously abnormal structure was observed in rat

关 键 词：麸炒 苍术 脾虚证 结肠组织通透性 紧密连接蛋白 信号通路 

分 类 号：R285.5[医药卫生—中药学]