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赵盼,钟缘,黎冬冬,龙达,彭清华.基于NLRP3/Caspase-1信号通路探讨除风益损汤对PM2.5诱导干眼的作用机制[J].湖南中医药大学学报,,():[点击复制] |
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| 基于NLRP3/Caspase-1信号通路探讨除风益损汤对PM2.5诱导干眼的作用机制 |
| 赵盼,钟缘,黎冬冬,龙达,彭清华 |
| (湖南中医药大学, 湖南 长沙 410208;湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心, 湖南 长沙 410208;上海市第六人民医院, 上海 200233) |
| 摘要: |
| 目的 探讨除风益损汤通过调控核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）/胱天蛋白酶-1（Caspase-1）信号通路对细颗粒物（PM2.5）诱导干眼小鼠模型的作用。方法 将小鼠随机分成空白组（NC组，10只）和造模组（30只），造模组予以双眼局部滴注PM2.5悬浮液（5 mg·mL-1）28 d进行造模，造模成功后再随机分为模型组（PM2.5组）、除风益损汤组（CFYSD组）、NLRP3抑制剂组（MCC950组），每组10只。CFYSD组予以除风益损汤（7.3 g·kg-1）灌胃，MCC950组予以MCC950（10 mg·kg-1）腹腔注射，NC组和PM2.5组予以等量蒸馏水灌胃。给药28 d后，检测各组小鼠泪液分泌量、泪膜破裂时间（TBUT）、角膜荧光染色评分，后进行取材。HE染色观察角膜组织病理变化，ELISA检测血清活性氧（ROS）、丙二醛（MDA）、超氧化物歧化酶2（SOD2）水平，Western blot及qPCR检测角膜组织NLRP3、Caspase-1、凋亡相关斑点样蛋白（ASC）、白细胞介素（IL）-1β、IL-18的蛋白及mRNA表达水平。结果 与NC组相比，PM2.5组泪液分泌量减少、TBUT缩短、角膜荧光染色评分增加（P<0.05）;血清ROS、MDA水平升高，SOD2水平下降（P<0.05）;角膜组织NLRP3、Caspase-1、ASC、IL-1β、IL-18的蛋白及mRNA表达水平升高（P<0.05）。与PM2.5组相比，CFYSD组和MCC950组泪液分泌量增加、TBUT延长、角膜荧光染色评分下降（P<0.05）;血清ROS、MDA水平下降，SOD2水平升高（P<0.05）;角膜组织NLRP3、Caspase-1、ASC、IL-1β、IL-18的蛋白及mRNA表达水平下降（P<0.05）。HE染色结果显示，NC组小鼠角膜组织各层结构清晰，未见异常;与NC组相比，PM2.5组小鼠角膜上皮细胞增生，细胞排列紊乱;与PM2.5组相比，MCC950组与CFYSD组小鼠在角膜结构完整性和细胞形态方面明显改善。结论 除风益损汤能增加PM2.5诱导干眼小鼠模型的泪液分泌量、延长TBUT、减轻角膜组织病理改变，其作用机制可能为抑制氧化应激过程及NLRP3/Caspase-1信号通路的激活，干预NLRP3炎症小体组装，减少IL-1β、IL-18等促炎性细胞因子的成熟与释放，从而调控炎症反应，最终缓解干眼症状。 |
| 关键词: 干眼 PM2.5 氧化应激 除风益损汤 核苷酸结合寡聚化结构域样受体蛋白3 胱天蛋白酶-1 |
| DOI： |
| 投稿时间：2026-03-20 |
| 基金项目:国家自然科学基金项目（81574031）；湖南省药食同源功能性食品工程技术研究中心开放基金项目（2018YSTY04）；湖南省教育厅科学研究项目（21A0238，22A0241，23A0300）；湖南中医药大学2021年“刘良院士工作站”指导项目（21YS002）。 |
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| Mechanism of action of Chufeng Yisun Decoction on PM2.5-induced dry eye via the NLRP3/Caspase-1 signaling pathway |
| ZHAO Pan,ZHONG Yuan,LI Dongdong,LONG Da,PENG Qinghua |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Engineering Technology Research Center for the Prevention and Treatment of Otorhinolaryngologic Diseases and Protection of Visual Function with Chinese Medicine, Changsha, Hunan 410208, China;Shanghai Sixth People's Hospital, Shanghai 200233, China) |
| Abstract: |
| Objective To investigate the effects of Chufeng Yisun Decoction on a mouse model of dry eye induced by fine particulate matter (PM2.5) through regulation of the NOD-like receptor thermal protein domain associated protein 3(NLRP3)/cysteine aspartate specific protease-1 (Caspase-1) signaling pathway. Methods Mice were randomly divided into blank group (NC group, 10 mice) and modeling group (30 mice). The modeling group received topical instillation of PM2.5 suspension (5 mg·mL-1) in both eyes for 28 d to establish the model. After successful modeling, the mice were randomly divided into model group (PM2.5 group), Chufeng Yisun Decoction group (CFYSD group), and NLRP3 inhibitor group (MCC950 group), with 10 mice in each group. The CFYSD group was administered Chufeng Yisun Decoction (7.3 g·kg-1) by gavage, the MCC950 group was administered MCC950 (10 mg·kg-1) by intraperitoneal injection, and the NC group and PM2.5 group were administered an equal volume of distilled water by gavage. After 28 d of administration, the tear secretion volume, tear film breakup time (TBUT), and corneal fluorescein staining score of mice in each group were measured, after which samples were collected. HE staining was used to observe pathological changes in corneal tissue; ELISA was used to determine serum levels of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase 2 (SOD2); and Western blot and qPCR were used to measure the protein and mRNA expression levels of NLRP3, Caspase-1, apoptosis-associated speck-like protein containing a CARD (ASC), interleukin (IL)-1β, and IL-18 in corneal tissue. Results Compared with the NC group, the PM2.5 group showed decreased tear secretion volume, shortened TBUT, and increased corneal fluorescein staining score (PM2.5.05); increased serum ROS and MDA levels and decreased SOD2 level (PM2.5.05); and increased protein and mRNA expression levels of NLRP3, Caspase-1, ASC, IL-1β, and IL-18 in corneal tissue (PM2.5.05). Compared with the PM2.5 group, the CFYSD group and MCC950 group showed increased tear secretion volume, prolonged TBUT, and decreased corneal fluorescein staining score (PM2.5.05); decreased serum ROS and MDA levels and increased SOD2 level (PM2.5.05); and decreased protein and mRNA expression levels of NLRP3, Caspase-1, ASC, IL-1β, and IL-18 in corneal tissue (PM2.5.05). HE staining results showed that the corneal tissue of mice in the NC group had clear structures in all layers with no abnormalities; compared with the NC group, mice in the PM2.5 group showed proliferation of corneal epithelial cells with disorganized cell arrangement; compared with the PM2.5 group, mice in the MCC950 group and CFYSD group showed marked improvement in corneal structural integrity and cell morphology. Conclusion Chufeng Yisun Decoction can increase tear secretion volume, prolong TBUT, and alleviate pathological changes in corneal tissue in a mouse model of PM2.5-induced dry eye. Its mechanism of action may involve inhibiting oxidative stress and activation of the NLRP3/Caspase-1 signaling pathway, interfering with NLRP3 inflammasome assembly, and reducing the maturation and release of pro-inflammatory cytokines such as IL-1β and IL-18, thereby regulating the inflammatory response and ultimately alleviating dry eye symptoms. |
| Key words: dry eye disease PM2.5 oxidative stress Chufeng Yisun Decoction NOD-like receptor thermal protein domain associated protein 3 cysteine aspartate specific protease-1 |
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