文献类型：期刊文献

中文题名：四神丸对脾肾阳虚型溃疡性结肠炎模型大鼠结肠组织PI3K/Akt/mTOR信号通路的免疫组化影响

英文题名：Immunohistochemical effect of sishen pill on PI3K/Akt/mTOR signal pathway in colonic tissue of rats with ulcerative colitis model of spleen kidney yang deficiency

作者：王燕[1];柳荣[1];朱向东[1]

第一作者：王燕

机构：[1]甘肃中医药大学,兰州730000

第一机构：甘肃中医药大学

年份：2021

卷号：29

期号：1

起止页码：42

中文期刊名：中国实验动物学报

外文期刊名：Acta Laboratorium Animalis Scientia Sinica

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金(81760833)。

语种：中文

中文关键词：溃疡性结肠炎;温肾健脾法;四神丸;磷脂酰肌醇-3-激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路;

外文关键词：ulcerative colitis;warming the kidney and strengthening the spleen method;Sishen pill;PI3K/Akt/mTOR signaling pathway;

摘要：目的本研究拟通过观察四神丸对脾肾阳虚型溃疡性结肠炎(UC)模型大鼠结肠组织病理特征和PI3K/Akt/mTOR信号通路相关蛋白免疫组化表达水平的影响,探讨溃疡性结肠炎发生的可能机制。方法120只SPF级Wistar大鼠(雌雄各半)随机分出20只作为空白组,其余100只作为造模组,造模采用DNBS/乙醇溶液灌肠+皮下注射氢化可的松+番泻叶灌胃法建立脾肾阳虚型UC大鼠模型。将成模大鼠随机分为5组,分别为模型组、美沙拉嗪组、四神丸高、中、低剂量组。模型组和空白组给予蒸馏水灌服,美沙拉嗪组按0.36 g/kg剂量灌胃,四神丸高、中、低剂量组分别按生药3.2、1.6、0.8 g/kg剂量灌胃,灌胃体积均为10 m L/kg。每日1次,连续21 d。采用HE染色法观察各组大鼠结肠组织病理改变,免疫组化法观察结肠组织PI3K p85、p-PI3K p85、AKT、p-AKT Ser473、mTOR、p-mTOR Ser2448蛋白的表达位置及表达水平。结果与空白组比较,模型组大鼠病理切片见肠黏膜部分消失,腺体消失,有大量的炎性细胞浸润,聚集于黏膜层和基层;PI3K p85、p-PI3K p85、AKT、p-AKT Ser473、mTOR、pmTOR Ser2448蛋白平均光密度值显著升高(P<0.01)。与模型组比较,药物组炎性细胞减少,黏膜层结构不同程度的恢复正常,美沙拉嗪组和四神丸中剂量组效果最好,黏膜结构接近空白对照组。四神丸低剂量组炎性细胞稍有减少,可见少量的腺体结构;四神丸高、中、低剂量组及美沙拉嗪组PI3K p85、p-PI3K p85、AKT、p-AKT Ser473、mTOR、p-mTOR Ser2448蛋白平均光密度值均有不同程度下降(P<0.01,P<0.05)。结论四神丸可能通过抑制PI3K/Akt/mTOR信号通路的激活改善脾肾阳虚型溃疡性结肠炎模型大鼠的肠黏膜损伤。
Objective This study observed the effects of Sishen pills on the pathological characteristics of colon tissue in spleen and kidney yang deficiency ulcerative coliti model rats and expression of PI3K/Akt/mTOR signaling pathway-related proteins to explore possible mechanisms of inflammation in an ulcerative colon.Methods A total of 120 SPF Wistar rats(half male and half female)were randomly assigned to a blank group of 20,and the other 100 rats were used as the model group.The rat model of ulcerative colitis with spleen-kidney Yang deficiency was established by DNBS/ethanol solution enema+hydrocortisone subcutaneous injection+senna leaf gavage.Rats with successful model establishment were randomly divided into five groups:model,mesalazine,Sishen pill high,Sishen pill medium,and Sishen pill low dose groups.Model and blank groups were administered distilled water.The mesalazine group was orally administered 0.36 g/kg mesalazine,and the high,middle and low dose groups of Sishen pills were orally administered 3.2,1.6,and 0.8 g/kg crude drug.Respectively,the volume of which was 10 m L/kg.Once a day for 21 d,colon tissues of rats were collected to observe general morphology and colon injury.HE staining was used to observe pathological changes.Immunohistochemistry was used to observe the localization and expression levels of PI3K p85,p-PI3K p85,AKT,p-AKT Ser473,mTOR,and p-mTOR Ser2448 proteins in colon tissues.Results Compared with the blank group,the intestinal mucosa had disappeared partially,glands had disappeared,and a large number of inflammatory cells had infiltrated and accumulated in mucosal and basal layers in pathological sections of the model group.The expression of PI3K p85,p-PI3K p85,AKT,p-AKT Ser473,mTOR,and p-mTOR Ser2448 proteins in the model group was increased significantly(P<0.01).Compared with the model group,inflammatory cells in the drug groups were reduced,and the structure of the mucosal layer returned to normal to varying degrees.Mesalazine and Sishen pill medium dose groups had the best effects,and the mucosal structure was close to that in the blank control group.Inflammatory cells in the low dose group of Sishen pill were slightly reduced,and a small number of glandular structures was seen.The expression of PI3K p85,p-PI3K p85,AKT,p-AKT Ser473,mTOR,and p-mTOR Ser2448 proteins was decreased to varying degrees in high,middle,low dose groups of Sishen pill and the mesalazine group(P<0.01,P<0.05).Conclusions Sishen pill may improve intestinal mucosal damage of ulcerative colitis model rats with spleen and kidney yang deficiency by inhibiting activation of the PI3K/Akt/mTOR signaling pathway.