基于PI3K/Akt/NF-κB通路探讨加味芍药汤对溃疡性结肠炎模型大鼠黏膜屏障修复的作用机制  

Exploration on the mechanism of Modified Shaoyao Decoction in repairing intestinal mucosal barrier of ulcerative colitis through PI3K/Akt/NF-κB pathway

作　　者：詹心雨 董蕾 徐慧妍 张海妹 贾永森 孔令玉 闫昕 Zhan Xinyu;Dong Lei;Xu Huiyan;Zhang Haimei;Jia Yongsen;Kong Lingyu;Yan Xin(College of Traditional Chinese Medicine,North China University of Science and Technology,Tangshan 063210,China;Department of Oncology,Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,China)

机构地区：[1]华北理工大学中医学院,唐山063210 [2]华北理工大学附属医院肿瘤科,唐山063000

出　　处：《国际中医中药杂志》2025年第2期196-204,共9页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81704059);河北省自然科学基金(H2022209057);华北理工大学研究生校级创新项目(2024S35)。

摘　　要：目的探讨加味芍药汤调控PI3K/Akt/NF-κB通路抗溃疡性结肠炎(UC)模型大鼠肠黏膜屏障损伤的作用机制。方法将72只Wistar大鼠按随机数字表法分为空白对照组、模型组、美沙拉嗪组和加味芍药汤高、中、低剂量组。除空白对照组,其余各组采用免疫复合法复制UC大鼠模型。造模成功后,美沙拉嗪组灌胃美沙拉嗪混悬液0.2 g/kg,加味芍药汤高、中、低组分别灌胃加味芍药汤水煎液44.5、22.3、11.1 g/kg,1次/d,共4周。采用HE染色观察结肠组织病理形态变化,进行疾病损伤指数(DAI)评分;免疫荧光法检测结肠组织紧密连接蛋白(ZO-1)、细胞骨架蛋白(F-actin)和血管内皮钙黏蛋白(VE-cadherin)表达;Western blot法检测结肠组织PI3K、Akt、NF-κB、VEGF、环氧化酶(COX-2)蛋白表达;qRT-PCR法检测结肠组织PI3K、Akt mRNA表达;ELISA法检测血清TNF-α、内皮素(ET)水平。结果与模型组比较,造模后第7、14、21、28天,美沙拉嗪组与加味芍药汤高剂量组DAI评分降低(P<0.05或P<0.01);美沙拉嗪组及加味芍药汤高、中、低剂量组结肠组织病理损伤评分降低(P<0.01),结肠组织ZO-1、F-actin、VE-cadherin蛋白免疫荧光强度升高(P<0.05或P<0.01),PI3K、Akt、NF-κB、VEGF蛋白表达降低(P<0.05或P<0.01),血清TNF-α、ET水平降低(P<0.05);美沙拉嗪组及加味芍药汤高、中剂量组COX-2蛋白表达降低,PI3K mRNA、Akt mRNA水平降低(P<0.05或P<0.01)。结论加味芍药汤可通过抑制PI3K/Akt/NF-κB通路拮抗UC模型大鼠肠屏障损伤,促进肠黏膜溃疡愈合。Objective To explore the mechanism of modified Shaoyao Decoction with Astragali Radix and Houttuynize Herba in regulating PI3K/Akt/NF-κB pathway against intestinal mucosal barrier injury in ulcerative colitis(UC)rats.Methods Totally 72 Wistar rats were randomly divided into a blank control group,a model group,a mesalazine group,and modified Shaoyao Decoction high-,medium-,and low-dose groups using a random number table method.After successful modeling,the mesalazine group was given 0.2 g/kg of mesalazine suspension by gavage,while modified Shaoyao Decoction high-,medium-,and low-dose groups were given 44.5,22.3,and 11.1 g/kg of modified Shaoyao Decoction by gavage,once a day,for a total of 4 weeks.The pathological changes of colon tissue were observed using HE staining;immunofluorescence assay was used to detect the expressions of tight junction protein(ZO-1),cytoskeletal protein(F-actin),and vascular endothelial cadherin(VE cadherin)in colon tissue;Western blot was used to detect the protein expressions of PI3K,Akt,NF-κB,VEGF,and cyclooxygenase(COX-2)in colon tissue;QRT PCR was used to detect the expressions of PI3K and Akt mRNA in colon tissue;ELISA method was used to detect the levels of serum TNF-αand endothelin(ET).Results Compared with the model group,the DAI scores of the mesalazine group and modified Shaoyao Decoction high-dosage group decreased significantly on the 7th,14th,21st and 28th day after modeling(P<0.05 or P<0.01).In mesalazine group and modified Shaoyao Decoction high-,medium-,and low-dose groups,the pathological injury score of colon tissue decreased(P<0.01),the immunofluorescence intensity of ZO-1,F-actin and VE-cadherin protein expression in colon tissue increased(P<0.05 or P<0.01),and the expressions of PI3K,Akt,NF-κB and VEGF protein decreased(P<0.01).The expression of COX-2 protein and the levels of PI3K mRNA and Akt mRNA decreased in the mesalazine group and modified Shaoyao Decoction high-and medium groups(P<0.05 or P<0.01).Conclusion Modified Shaoyao Decoction can antagonize the i

关 键 词：结肠炎 溃疡性 PI3K/Akt/NF-κB通路 加味芍药汤 肠黏膜屏障 大鼠 

分 类 号：R73[医药卫生—肿瘤]