川芎嗪通过抑制大鼠角膜碱烧伤诱导的TLR4/NF-κB信号通路激活及NLRP1/NLRP3炎性小体，从而抑制角膜炎和新生血管形成。
Tetramethylpyrazine inhibits keratitis and neovascularization induced by corneal alkali burn by suppressing the TLR4/NF-κB pathway activation and NLRP1/NLRP3 inflammasomes in rats.
作者信息
Jiang Tao, Wang Chun-Yang, Chen Ying
机构信息
Department of Ophthalmology, Shaoxing Campus, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Shaoxing, Zhejiang, 312300, China; Department of Ophthalmology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine (Key Laboratory for Corneal Diseases Research of Zhejiang Province), Hangzhou, Zhejiang, 310016, China.
Department of Ophthalmology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310009, China.
出版信息
Exp Eye Res. 2023 Oct 28:109704. doi: 10.1016/j.exer.2023.109704.
The role and related mechanisms of tetramethylpyrazine (TMP) in corneal alkali burn in rats were expected to be explored in this article. After construction of corneal alkali burn rat models, TMP eye drops were given four times daily for consecutive 7 days. H&E staining was utilized for observing the histopathological changes of corneas on the 3rd and 7th days of treatment; immunohistochemistry for detecting the Nestin protein expression changes; qRT-PCR for determining the expression changes of genes correlated with neovascularization [C-X-C Motif Chemokine Ligand 1 (CXCL-1), vascular endothelial growth factor A (VEGFA) and CD31] and inflammation-related factors [monocyte chemoattractant protein-1 (MCP-1), interleukin-1β (IL-1β), tumour necrosis factor α (TNF-α), and IL-6]; Western blot for testing NLR Family Pyrin Domain Containing 1 (NLRP1)/NLRP3 inflammasomes and toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) pathway-related protein expression changes. All above trials were completed based on rat corneal tissue. TMP ameliorated the pathological damage in alkali-burned rat corneal tissue. Specifically, TMP treatment decreased Nestin-positive cell expression and the CXCL-1, VEGFA and CD31 mRNA expression in alkali-burned rat corneal tissue dose-dependently. TMP also down-regulated the IL-1β, TNF-α, MCP-1 and IL-6 mRNA expression and inhibited the NLRP1, caspase-1, NLRP3, pro-IL-1β and mature IL-1β protein expression in the alkali-burned rat corneal tissue. In addition, TMP treatment down-regulated the myeloid differentiation factor 88 (MyD88) and TLR4 protein expression and decreased the p-NF-κB p65/NF-κB p65 ratio in the alkali-burned rat corneal tissue. The mechanism of TMP relieving the inflammatory response and inhibiting neovascularization caused by corneal alkali burn in rats might have a correlation with the suppression of acitivation of TLR4/NF-κB pathway and NLRP1/NLRP3 inflammasomes in rats.
本文旨在探讨川芎嗪（TMP）在大鼠角膜碱烧伤中的作用及相关机制。构建大鼠角膜碱烧伤模型后，每天给予TMP滴眼液4次，连续7天。采用苏木精-伊红（H&E）染色观察治疗第3天和第7天角膜的组织病理学变化；免疫组织化学检测巢蛋白（Nestin）蛋白表达变化；实时定量聚合酶链反应（qRT-PCR）测定与新生血管形成相关基因[C-X-C基序趋化因子配体1（CXCL-1）、血管内皮生长因子A（VEGFA）和CD31]及炎症相关因子[单核细胞趋化蛋白-1（MCP-1）、白细胞介素-1β（IL-1β）、肿瘤坏死因子α（TNF-α）和IL-6]的表达变化；蛋白质免疫印迹法检测含NLR家族吡啶结构域蛋白1（NLRP1）/NLRP3炎性小体及Toll样受体4（TLR4）/核因子κB（NF-κB）通路相关蛋白表达变化。所有上述实验均基于大鼠角膜组织完成。TMP改善了碱烧伤大鼠角膜组织的病理损伤。具体而言，TMP治疗剂量依赖性地降低了碱烧伤大鼠角膜组织中Nestin阳性细胞表达以及CXCL-1、VEGFA和CD31 mRNA表达。TMP还下调了碱烧伤大鼠角膜组织中IL-1β、TNF-α、MCP-1和IL-6 mRNA表达，并抑制了NLRP1、半胱天冬酶-1（caspase-1）、NLRP3、前体IL-1β和成熟IL-1β蛋白表达。此外，TMP治疗下调了碱烧伤大鼠角膜组织中髓样分化因子88（MyD88）和TLR4蛋白表达，并降低了p-NF-κB p65/NF-κB p65比值。TMP减轻大鼠角膜碱烧伤所致炎症反应并抑制新生血管形成的机制可能与抑制大鼠TLR4/NF-κB通路及NLRP1/NLRP3炎性小体的激活有关。
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