肠愈宁对溃疡性结肠炎大鼠PI3K/AKT信号通路作用机制的研究  

Mechanism of Changyuning on PI3K/AKT Signaling Pathway in Ulcerative Colitis Rats

作　　者：王海强[1] 张萌[2] 熊丽 马晨曦 王瑶[1] 郑丽红[3] Wang Haiqiang;Zhang Meng;Xiong Li;Ma Chenxi;Wang Yao;Zheng Lihong(First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China;The First Clinical College of Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China;The Fourth Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin 150001,China)

机构地区：[1]黑龙江中医药大学附属第一医院,哈尔滨150040 [2]黑龙江中医药大学第一临床医学院,哈尔滨150040 [3]黑龙江中医药大学附属第四医院,哈尔滨150001

出　　处：《世界科学技术-中医药现代化》2023年第11期3736-3743,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：黑龙江省自然科学基金委员会面上项目(H2016077):肠愈宁颗粒对溃疡性结肠炎大鼠钙卫蛋白影响的相关研究,负责人:王海强;黑龙江中医药大学科研基金面上项目(2019MS02):基于PI3K/AKT信号通路研究肠愈宁颗粒对溃疡性结肠炎大鼠Bcl-2、Bax、Caspase-3蛋白表达的影响,负责人:王瑶;2020年黑龙江中医药大学研究生创新科研项目(2020yjscx014):基于PI3K/AKT信号通路研究肠愈宁颗粒对溃疡性结肠炎大鼠PTEN、TLR2、Caspase-3、Caspase-9蛋白表达的影响,负责人:王海强

摘　　要：目的探讨肠愈宁方对2,4,6-三硝基苯磺酸(TNBS)诱导的溃疡性结肠炎(UC)大鼠结肠组织磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/AKT)信号通路中PTEN、TLR2、Caspase-3、Caspase-9蛋白的作用机制。方法将70只SPF级大鼠适应性喂养7天后分为空白组10只和造模组60只,造模组大鼠采用TNBS/乙醇溶液灌肠诱导UC模型。将成功建立模型的大鼠随机分为模型组,美沙拉嗪组,肠愈宁高、中、低剂量组、肠愈宁高、中、低剂量组分别灌服相当于生药量为4 g·mL^(-1)、2 g·mL^(-1)、1 g·mL^(-1)的肠愈宁方溶液10 mL·kg^(-1),美沙拉嗪组灌服相当于生药量为0.2 g·kg^(-1)美沙拉嗪悬浊液10 mL·kg^(-1),空白对照组、模型组给予等量10 mL·kg^(-1)生理盐水灌胃。各组大鼠灌胃均每天1次,连续14天。末次给药后处死大鼠取材。评估疾病活动指数,HE染色观察结肠变化,进行结肠组织损伤指数评分,蛋白免疫印迹法检测结肠组织PI3K、AKT、PTEN、TLR2、Caspase-3、Caspase-9蛋白的表达。结果与空白组比较,模型组PI3K、AKT、Caspase-3、Caspase-9及TLR2蛋白表达水平增高(P<0.05),PTEN蛋白表达水平降低(P<0.05)。与模型组比较,肠愈宁高剂量组PI3K、AKT、Caspase-3、Caspase-9、TLR2蛋白表达水平均明显下降(P<0.05),PTEN蛋白表达水平显著升高(P<0.05);美沙拉嗪组、肠愈宁中剂量PI3K、AKT、Caspase-3水平均降低(P<0.05),PTEN蛋白表达水平升高(P<0.05),在Caspase-9中表达无统计学意义;肠愈宁低剂量组在各蛋白表达情况与模型组无统计学意义。结论肠愈宁方治疗UC作用显著,其机制可能与调控PI3K/AKT信号通路及其相关蛋白PTEN、TLR2、Caspase-3、Caspase-9有关,能够有效缓解UC大鼠症状,减轻结肠病理损伤。Objective To investigate the effect of Changyuning prescription on PTEN,TLR2,Caspase-3,Caspase-9 in phosphatidylinositol-3 kinase/protein kinase B(PI3K/AKT)signaling pathway in colon tissue of 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced ulcerative colitis(UC)rats.Methods After adaptive feeding for 7 days,70 SPF rats were divided into blank group of 10 and modeling group of 60.The rats in the modeling group were given TNBS/ethanol solution enema to induce UC model.The rats that successfully bulit the model were randomly divided into model group,mesalazine group,Changyuning high-dose,medium-dose and low-dose groups,and Changyuning high-dose,medium-dose and low-dose groups,respectively,and the equivalent amount of crude drug was 4 g·mL^(-1),2 g·mL^(-1),1 g·mL^(-1)of Changyuning prescription solution 10 mL·kg^(-1),the mesalamine group was given 10 mL·kg^(-1)of mesalamine suspension equivalent to 0.2 g·kg^(-1)crude drug,the blank control group and model group were given an equal volume of 10 mL·kg^(-1)of normal saline was given by gavage.The rats in each group were given intragastric administration once a day for 14 consecutive days.Rats were sacrificed after the last administration.Disease activity index was evaluated;colon changes were observed by HE staining;colon tissue damage index was evaluated;The expression of PI3K,AKT,PTEN,TLR2,Caspase-3,and Caspase-9 proteins was detected by Western Blotting in colon tissue.Results Compared with the blank group,the protein expression levels of PI3K,AKT,Caspase-3,Caspase-9 and TLR2 in the model group were increased(P<0.05),and the protein expression level of PTEN was decreased(P<0.05).Compared with the model group,the expression levels of PI3K,AKT,Caspase-3,Caspase-9 and TLR2 proteins in the Changyuning high-dose group were significantly decreased(P<0.05),and the protein expression levels of PTEN were significantly increased(P<0.05);The levels of PI3K,AKT and Caspase-3 in the salazine group and Changyuning medium dose were decreased(P<0.05),the expression level of

关 键 词：溃结性结肠炎 肠愈宁方 PI3K/AKT信号通路 PTEN TLR2 Caspase-3 CASPASE-9 

分 类 号：R285.5[医药卫生—中药学]