石斛酚抑制内质网应激减轻大鼠角膜碱烧伤实验研究  

Dendrobine inhibits endoplasmic reticulum stress to alleviate cornea alkali burn in rats

作　　者：朱吉凤 孔超 彭敏 姚智华 ZHU Jifeng;KONG Chao;PENG Min;YAO Zhihua(Department of Burns and Plastic Surgery,the Affiliated Yongchuan Hospital of Chongqing Medical University,Chongqing 402160,China;Department of Plastic Surgery,the First Affiliated Hospital of Army Medical University,Chongqing 400038,China)

机构地区：[1]重庆医科大学附属永川医院烧伤整形科,重庆402160 [2]陆军军医大学附属第一医院整形外科,重庆400038

出　　处：《陕西医学杂志》2025年第5期590-595,601,共7页Shaanxi Medical Journal

基　　金：重庆市基础与前沿研究计划项目(Y2021094201)。

摘　　要：目的:探讨石斛酚对大鼠角膜碱烧伤的改善作用及可能机制。方法:将64只大鼠随机分为对照组(15只)和造模组(49只),通过氢氧化钠烧灼大鼠右眼角膜中央构建角膜碱烧伤模型。从造模成功的角膜碱烧伤大鼠中随机取45只,分为模型组和石斛酚低、高剂量组,每组15只。建模后,石斛酚低、高剂量组大鼠在距角膜缘1 mm处分4个象限结膜下分别注射浓度为100、300 g/ml石斛酚20μl;对照组及模型组注射等量0.9%氯化钠溶液,均连续给药14 d。裂隙灯下观察角膜新生血管情况并计算新生血管面积;HE染色观察大鼠角膜组织损伤情况;ELISA法检测眼内房水中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平;免疫组织化学染色检测大鼠角膜组织中血管内皮生长因子(VEGF)、MMP-9蛋白表达;RT-qPCR及Western blot检测大鼠角膜组织中转录因子剪接型X-盒结合蛋白1(XBP1s)、活化转录因子4(ATF4)、ATF6、免疫球蛋白结合蛋白(BiP)、EBP同源蛋白(CHOP)mRNA及蛋白表达。结果:建模后4、7、14 d,模型组和石斛酚低、高剂量组大鼠角膜均可见新生血管,且石斛酚低、高剂量组大鼠角膜新生血管长度及面积小于模型组(均P<0.05)。模型组大鼠角膜可见较多新生血管,管腔内含有较多红细胞,并可见大量炎症细胞浸润;与模型组比较,石斛酚低、高剂量组大鼠角膜的上述组织病理学损伤减轻。与模型组比较,石斛酚低、高剂量组大鼠眼内房水TNF-α、IL-1β水平,角膜组织VEGF、MMP-9阳性表达以及XBP-1s、ATF4、ATF6、BiP、CHOP mRNA及蛋白表达水平降低(均P<0.05)。石斛酚高剂量组以上指标优于石斛酚低剂量组(均P<0.05)。结论:石斛酚能够改善角膜碱烧伤大鼠的角膜炎症反应,减少角膜血管新生,从而减轻碱烧伤引起的角膜损伤,其作用机制可能与抑制内质网应激有关。Objective:To investigate the ameliorative effects of dendrobine on cornea alkali burn in rats and its potential mechanism.Methods:A total of 64 rats were randomly divided into a control group(15 rats)and a modeling group(49 rats).A corneal alkali burn model was established by applying sodium hydroxide to the central cornea of the right eye in rats.From the successfully modeled rats,45 were randomly selected and divided into a model group and two dendrobine groups(low and high doses),with 15 rats in each group.After modeling,rats in the low-and high-dose dendrobine groups received sub conjunctival injections of 20μl dendrobine at concentrations of 100 and 300 g/ml respectively,in four quadrants 1 mm from the limbus.The control and model groups received equal volumes of 0.9%sodium chloride solution.All treatments were administered daily for 14 days.Corneal neovascularization was observed under a slit-lamp and the neovascular area was calculated.Corneal tissue damage was assessed by HE staining.Levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in the aqueous humor were measured by ELISA.Expression of vascular endothelial growth factor(VEGF)and matrix metalloproteinase-9(MMP-9)proteins in corneal tissues was detected by immunohistochemical staining.Expression of transcription factors spliced X-box binding protein 1(XBP1s),activating transcription factor 4(ATF4),ATF6,immunoglobulin binding protein(BiP)and EBP homologous protein(CHOP)at the mRNA and protein levels in corneal tissues was detected by RT-qPCR and Western blot.Results:At 4,7 and 14 days after modeling,corneal neovascularization observed in rats of the model and dendrobine groups,with shorter neovascular length and smaller area in the low-and high-dose dendrobine groups compared to the model group(all P<0.05).The model group showed more abundant neovascularization with red blood cells in the lumen and extensive inflammatory cell infiltration.Histopathological damage in the corneal tissues reduced in the low-and high-dose dendrobine group

关 键 词：角膜碱烧伤 石斛酚 内质网应激 炎症反应 血管新生 大鼠 

分 类 号：R-33[医药卫生]