基于TRAF6/TAK1信号通路探讨清眩润目饮对干眼大鼠的作用机制  

Exploring the Mechanism of Qingxuan Runmu Decoction on Rats with Dry Eye Based on the TRAF6/TAK1 Signaling Pathway

作　　者：赵珊珊 刘颖[1] 王佳娣[2] 姚靖[2] Zhao Shanshan;Liu Ying;Wang Jiadi;Yao Jing(Heilongjiang University of Chinese Medicine,Heilongjiang,Harbin 150000,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150000 [2]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150000

出　　处：《中国中医急症》2024年第11期1920-1923,1946,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973908);黑龙江省中医药经典普及化专项课题项目(ZYW2022-051)。

摘　　要：目的探究清眩润目饮通过TRAF6/TAK1信号通路对干眼模型大鼠的作用机制。方法将40只雌性大鼠分为空白组、模型组、清眩润目饮组、玻璃酸钠组,模型组、清眩润目饮组、玻璃酸钠组予0.2%BAC滴眼建立干眼模型。造模成功后,清眩润目饮组予中药灌胃+PBS缓冲液滴眼;玻璃酸钠组大鼠予等量生理盐水灌胃+玻璃酸钠滴眼液滴眼,连续2周。给药2周时进行泪液分泌检测(SIT)及角膜荧光素钠评分(FL),处死大鼠并留取角膜组织,采用ELISA检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)及MMP9含量,HE染色观察角膜组织病理学变化;免疫组化染色观察TFAF6的蛋白表达,Western blotting检测TRAF6、p-TAK1及TAK1蛋白的表达量。结果与模型组相比,清眩润目饮组及玻璃酸钠组SIT显著增高(P<0.05),FL显著降低(P<0.05)。HE染色结果显示,清眩润目饮组及玻璃酸钠组角膜组织形态得到明显改善。免疫组化结果显示,清眩润目饮组及玻璃酸钠组TFAF6含量显著低于模型组(P<0.05)。Western blotting结果显示,与模型组相比,清眩润目饮组及玻璃酸钠组大鼠TRAF6水平降低(P<0.05),清眩润目饮组p-TAK1/TAK1水平显著降低(P<0.05)。ELISA结果显示,清眩润目饮组及玻璃酸钠组IL-1β、TNF-α及MMP9水平较模型组显著降低(P<0.05)。结论清眩润目饮能够改善干眼大鼠角膜组织损伤,抑制炎症反应的发生,从而缓解干眼大鼠眼部症状,其机制可能是通过抑制TRAF6/TAK1信号通路实现的。Objective:To explore the mechanism of action of Qingxuan Runmu Decoction(QRD)on dry eye model rats through the TRAF6/TAK1 signaling pathway.Methods:40 female rats were divided into blank group,model group,QRD group,and sodium hyaluronate group.Model group,QRD group,and sodium hyaluronate group were treated with 0.2%BAC eye drops for two weeks.After successful modeling,QRD group was administered QRD and PBS buffer eye drops for two weeks;Sodium hyaluronate group was given an equal amount of physiological saline by gavage and sodium hyaluronate eye drops.After 2 weeks of administration,SIT and FL scores were measured,rats were euthanized and corneal tissue was collected.ELISA was used to detect IL-1β,TNF-α,and MMP9 levels;HE staining was used to observe pathological changes in corneal tissue;Immunohistochemical staining was used to observe the protein expression of TFAF6,and Western blotting was used to detect the expression levels of TRAF6,p-TAK1,and TAK1 proteins.Results:Compared with model group,SIT in QRD group,and sodium hyaluronate group significantly increased(P<0.05),and FL significantly decreased(P<0.05);The HE staining results showed that the corneal tissue morphology of QRD group and sodium hyaluronate group was improved considerably;the immunohistochemical results showed that the TFAF6 contents in QRD group,and sodium hyaluronate group were significantly lower than that in the model group(P<0.05);Western blotting results showed that compared with model group,the levels of TRAF6 of QRD group,and sodium hyaluronate group were reduced(P<0.05),and p-TAK1/TAK1 of QRD group was reduced(P<0.05);ELISA results showed that the levels of IL-1β,TNF-α,and MMP9 in QRD group,and sodium hyaluronate group were lower than those in model group(P<0.05).Conclusion:QRD can improve corneal tissue damage in rats with dry eye,inhibit the occurrence of inflammatory reactions,and alleviate eye symptoms.Its mechanism may be achieved by inhibiting the FRAF6/TAK1 signaling pathway.

关 键 词：干眼 清眩润目饮 TRAF6/TAK1信号通路 炎症 大鼠 

分 类 号：R777.34[医药卫生—眼科]