摘要
目的研究刺槐素对干眼症(DED)大鼠的保护作用及可能的分子机制。方法60只SD大鼠随机分为空白组(不做处理);模型组;0.1%SH组;刺槐素0.1%、0.25%、0.5%组。每组10只,除正常组外,每日4次皮下注射12.5 mg/天氢溴酸东莨菪碱(SCOP),共7天。所有的药物均为在眼球表面局部给药,每只眼睛20μL,每天4次。通过泪液分泌实验检测各组大鼠泪液分泌情况,通过角膜荧光染色检测各组大鼠角膜表面缺损情况,过碘酸雪夫试剂(PAS)染色观察结膜杯状细胞数量,酶联免疫吸附法(ELISA)检测各组大鼠角膜组织中炎症反应和氧化应激相关指标含量。免疫印迹法(Western blot)检测各组大鼠角膜组织中Toll样受体4(TLR4)通路相关蛋白TLR4、髓样分化因子88(MyD88)、核因子κB p65(NF-κB p65)、Pyrin域蛋白3(NLRP3)蛋白表达。结果与正常组相比,模型组大鼠泪液分泌量、结膜杯状细胞数量明显减少,角膜荧光素染色评分显著升高,但角膜新血管生成评分减少,同时角膜组织中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-10、丙二醛(MDA)水平上调,而超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)含量则下调,TLR4、MyD88、NLRP3、p-NF-κB p65蛋白相对表达量均升高(P<0.05);与模型组相比,经刺槐素0.25%、0.5%组治疗后,大鼠泪液分泌量、结膜杯状细胞数量明显增加,角膜荧光素染色评分显著降低,角膜新血管生成评分显著增加,同时角膜组织中IL-1β、TNF-α、IL-10、MDA水平下调,而SOD和GSH-Px含量则上调(P<0.05),尤其是刺槐素0.5%组上述指标变化更明显,而且角膜组织中TLR4、MyD88、NLRP3、p-NF-κB p65蛋白相对表达量较模型组显著降低(P<0.05)。结论刺槐素对干眼症大鼠具有保护作用,其保护机制与抑制干眼症大鼠的氧化应激反应、炎症反应和TLR4/MyD88/NLRP3通路活性有关。
Objective To study the protective effect of acacetin on dry eye(DED)rats and further to study its possible mechanism.Methods Sixty SD rats were randomly divided into control group,model group,0.1%of Sodium hyaluronate(0.1%SH)group,acacetin 0.1%,0.25%,0.5%group,with 10 rats in each.Except the control group,12.5 mg·d-1Scopolamine hydrobromide(SCOP)was injected subcutaneously four times a day for 7 days.All the drugs were administered locally on the surface of the eyeball,20μL in each eye,four times a day.DED rat model was established by lacrimal gland removal and schirmer test,and corneal fluorescein staining were performed to evaluate the effects of acacetin on the ocular surface.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of inflammatory response and oxidative stress related indexes in corneal tissues of rats in each group.Western blot was used to detect the expression of Toll-like receptor 4(TLR4)signaling pathway related proteins,such as TLR4,myeloid differentiation factor 88(MyD88),nuclear factor kappa B p65(NF-κB p65),Pyrin domain containing protein 3(NLRP3)in corneal tissue.Results Compared with the control group,the volume of tear secretion and the number of conjunctival goblet cells in the model group decreased significantly,and the corneal fluorescein staining score increased significantly,but the corneal neovascularization score decreased,while the levels of Interleukin 1 beta(IL-1β),tumor necrosis factor alpha(TNF-α),IL-10,malondialdehyde(MDA)in the corneal tissue were lower and the levels of superoxide dismutase(SOD);glutathione peroxidase(GSH-Px)were increased;the relative expression of TLR4,MyD88,NLRP3,p-NF-κB p65 protein were all increased(P<0.05).Compared with the model group,after treatment with 0.25%and 0.5%of acacetin,the amount of tear secretion and the number of goblet cells in conjunctiva were significantly increased;the corneal fluorescein staining score decreased significantly and the corneal neovascularization score increased;the levels of IL-1β,TNF-α,IL-10 and MDA in corneal tissues decreased,while the levels of SOD and GSH-Px increased(P<0.05),especially in acacetin 0.5%group,in which the relative expressions of TLR4,MyD88,NLRP3,p-NF-κB p65protein in cornea tissues were significantly lower than that in model group(P<0.05).Conclusion Acacetin has protective effect on xerophthalmia rats,and its protective mechanism is related to the inhibition of oxidative stress response,inflammatory response and TLR4/MyD88/NLRP3 pathway activity in xerophthalmia rats.
作者
白雪
刘艳茹
王海
Bai Xue;Liu Yanru;Wang Hai(Sanya Hospital of Traditional Chinese Medicine,Sanya 572000,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第7期2740-2747,共8页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
关键词
刺槐素
TLR4通路
干眼症
炎症反应
氧化应激
Acacetin
TLR4 pathway
Xerophthalmia
Inflammatory response
Oxidative stress
作者简介
通讯作者:白雪,主治中医师,主要研究方向:眼科临床疾病的中西医结合治疗。
