基于Keap1-Nrf2-ARE途径探讨铁皮石斛多糖抗矽肺纤维化的机制  

Exploring the mechanism of Dendrobium officinale polysaccharide against silicosis fibrosis based on Keap1-Nrf2-ARE pathway

作　　者：郭建宏 罗菲 窦玉玉 王谨莹 张旭清 唐汉庆[2] Guo Jianhong;Luo Fei;Dou Yuyu;Wang Jinying;Zhang Xuqing;Tang Hanqing(Graduate School,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;School of Basic Medicine,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院研究生学院,广西百色533000 [2]右江民族医学院基础医学院,广西百色533000

出　　处：《右江民族医学院学报》2023年第3期367-372,383,共7页Journal of Youjiang Medical University for Nationalities

基　　金：国家自然科学基金资助项目(81760759)。

摘　　要：目的观察铁皮石斛多糖(DOP)通过Keap1-Nrf2-ARE信号通路抗矽肺纤维化的效果,探讨其可能的机制。方法将60只BALB/c小鼠按随机数字表法随机分为对照组、模型组、吡非尼酮(PFD)组、DOP低剂量组、DOP中剂量组、DOP高剂量组。除对照组外,其余组小鼠采用口咽法滴入50.0 mg/mL SiO_(2)混悬液200μL造模,之后等待4周。第5周开始PFD组每日灌胃吡非尼酮,DOP低剂量组(125 mg/kg)、DOP中剂量组(250 mg/kg)、DOP高剂量组(500 mg/kg)每日分别灌胃对应剂量的DOP,对照组和模型组则每日灌胃等量的生理盐水,连续给药4周后进行相关检测。HE和天狼星红染色检测小鼠肺组织的病理变化和纤维化程度;ELISA检测肺组织TGF-β1的含量;qRT-PCR检测肺组织Nrf2、Keap1 mRNA表达;Western Blot检测肺组织Nrf2、Keap1蛋白表达。结果与对照组相比,模型组小鼠肺组织TGF-β1含量明显升高,肺组织炎性细胞浸润明显,大量胶原纤维沉积;肺组织Nrf2、Keap1 mRNA及相应的蛋白表达均呈上升趋势。与模型组相比,4个给药组小鼠肺组织炎性细胞浸润及纤维化程度均有不同程度的改善;肺组织Nrf2、Keap1 mRNA及相应的蛋白表达水平均有不同程度的降低。结论DOP可以通过下调Keap1-Nrf2-ARE信号通路上的相关分子的表达水平,从而有效改善矽肺纤维化。Objective To observe the effect of Dendrobium officinale polysaccharide(DOP)on silicosis fibrosis through the Keap1-Nrf2-ARE signaling pathway and explore its underlying mechanism.Methods Sixty BALB/c mice were randomly assigned to control group,model group,pirfenidone(PFD)group,low-dose DOP group,medium-dose DOP group,and high-dose DOP group using a random number table.Except for the control group,the mice in the other groups were orally administered with 200μL of 50.0 mg/mL SiO_(2)suspension to induce silicosis fibrosis.Starting from the fifth week,the PFD group received daily oral administration of pirfenidone,while the low-dose(125 mg/kg),medium-dose(250 mg/kg),and high-dose(500 mg/kg)DOP groups received corresponding doses of DOP.The control and model groups were administered equal amounts of normal saline daily.After four weeks of treatment,lung tissue samples were collected for analysis.Pathological changes and fibrosis degree were assessed using HE and Sirius red staining.The content of TGF-β1 in lung tissue was measured by ELISA.The mRNA expression levels of Nrf2 and Keap1 in lung tissue were determined by qRT-PCR,and the protein expressions of Nrf2 and Keap1 were examined using Western Blotting.Results Compared to the control group,the model group exhibited significantly increased TGF-β1 content,noticeable inflammatory cell infiltration,and extensive collagen fiber deposition in the lung tissue.The mRNA and protein expressions of Nrf2 and Keap1 were up-regulated.In contrast,the administration groups showed varying degrees of improvement in inflammatory cell infiltration and fibrosis.Additionally,the expression levels of Nrf2,Keap1 mRNA,and corresponding proteins in the lung tissue were reduced to varying degrees compared to the model group.Conclusion DOP effectively improves silicosis fibrosis by down-regulating the expression of relevant molecules in the Keap1-Nrf2-ARE signaling pathway.

关 键 词：矽肺 铁皮石斛多糖 肺纤维化 Keap1-Nrf2-ARE通路 

分 类 号：R563.13[医药卫生—呼吸系统]