基于TLR4/MyD88/NF-κB信号通路研究清眩润目饮对苯扎氯铵诱导的干眼大鼠的作用机制  

The Mechanism of Qingxuan Runmu Yin(清眩润目饮)on Benzalkonium Chloride-Induced Dry Eye in Rats Based on TLR4/MyD88/NF-κB Signaling Pathway

作　　者：赵珊珊 王佳娣[2] 刘颖[1] 姚靖[2] ZHAO Shanshan;WANG Jiadi;LIU Ying;YAO Jing(Heilongjiang University of Chinese Medicine,Harbin Heilongjiang 150000,Chine;First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin Heilongjiang 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：《中医药导报》2024年第8期27-33,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

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:基于TLR4/MyD88/NF-κB信号通路探讨清眩润目饮对苯扎氯铵(BAC)诱导干眼模型大鼠的作用机制。方法:将40只SD大鼠随机分为空白组、模型组、清眩润目饮组、玻璃酸钠组,每组10只(20眼)。除空白组外,各组大鼠采用0.2%BAC滴眼,5μL/眼,2次/d,连续14 d,制备大鼠干眼模型。造模后,清眩润目饮组予中药灌胃,同时予磷酸盐缓冲液滴眼;玻璃酸钠组大鼠予等量生理盐水灌胃及玻璃酸钠滴眼液滴眼;空白组和模型组予等量生理盐水灌胃及磷酸盐缓冲液滴眼,2次/d,连续14 d。末次给药1 h后对各组大鼠行泪液分泌量检测(SIT)及角膜荧光素钠染色(FL)评分;透射电镜观察角膜细胞超微结构;HE染色观察结膜组织病理学变化;RT-qPCR检测各组大鼠角膜、结膜及泪腺中TLR4 mRNA、MyD88 mRNA、NF-κB p65 m RNA的表达;ELISA检测各组大鼠角膜、结膜及泪腺中磷酸化p65(p-p65)蛋白的表达。结果:造模14 d后,模型组、清眩润目饮组及玻璃酸钠组大鼠SIT低于空白组(P<0.01),FL评分高于空白组(P<0.01)。给药14 d后,清眩润目饮组、玻璃酸钠组大鼠SIT均高于模型组(P<0.01或P<0.05),FL评分均低于模型组(P<0.05)。透射电镜显示模型组大鼠角膜间隙增大,面积增宽,排列不规则;清眩润目饮组及玻璃酸钠组大鼠角膜上皮细胞间隙增宽程度减轻,排列较规则,角膜结构改善。HE染色显示,与模型组比较,清眩润目饮组及玻璃酸钠组大鼠结膜上皮细胞形态及排列恢复,杯状细胞数量上升。模型组大鼠角膜、结膜及泪腺中TLR4 mRNA、MyD88 mRNA相对表达量高于空白组(P<0.01);清眩润目饮组及玻璃酸钠组大鼠角膜、结膜及泪腺中TLR4 mRNA、MyD88 mRNA相对表达量均低于模型组(P<0.01);清眩润目饮组大鼠角膜中TLR4 mRNA及结膜、泪腺中MyD88 mRNA相对表达量均低于玻璃酸钠组(P<0.01)。模型组大鼠角膜、结膜及泪腺中p-p65蛋白表达水平均高于空白组(P<0.01);清眩润�Objective:To evaluate the mechanism of Qingxuan Runmu Yin in the benzalkonium chloride (BAC)-induced dry eye model in rats based on the TLR4/MyD88/NF-κB signaling pathway. Methods: Totally 40 SD rats were randomely divided into blank group, model group, Qingxuan Runmu Yin (QRY) group and sodium hyaluronate group, with 10 rats (20 eyes) in each group. 0.2% BAC eye drops were administered to each group of rats excluding the blank group, 5 μL/eye, twice a day for 14 days, to prepare a dry eye model of rats with excessive evaporation. After modeling, the rats in the QRY group were given QRY by gavage and phosphate buffer drops. The rats in the sodium hyaluronate group were given equal amounts of saline by gavage and sodium hyaluronate eye drops. The rats in the blank and model groups were administrated with equal amounts of saline and phosphate buffers twice a day for a 14 days. Schirmer Test (SIT) and corneal sodium fluorescein (FL) staining scores of rats in each group were detected 1 hour after the last administration. Transmission electron microscopy was used to examine the ultrastructure of corneal cells. HE staining was performed to view the histopathologic changes in the conjunctiva. RT-qPCR was used to detect the expression of TLR4 mRNA, MyD88 mRNA, NF-κB p65 mRNA in the cornea, conjunctiva and lachrymal glands of rats in each group. ELISA was used to detect the expression of phosphorylated p-p65 protein in the cornea, conjunctiva and lacrimal gland of rats. Results: After 14 days of modeling, model group, QRY group and sodium hyaluronate group showed lower SIT than blank group (P<0.01), while higher FL score than blank group (P<0.01). After 14 days of administration, QRY group and sodium hyaluronate group showed significantly higher SIT than model group (P<0.01 or P<0.05), while lower FL scores than model group (P<0.05). Transmission electron microscopy showed that the corneal epithelial cell gap was widened and irregularly arranged in the model group, while the cell gap widening was reduced and more regu

关 键 词：干眼 清眩润目饮 TLR4/MyD88/NF-κB信号通路 苯扎氯铵 大鼠 

分 类 号：R285.5[医药卫生—中药学]