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刘溪源,喻京生.祛风明目丸对实验性自身免疫性葡萄膜炎大鼠NLRP3、ASC、IL-1β表达的影响[J].湖南中医药大学学报英文版,2023,43(10):1779-1785.[Click to copy
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| 祛风明目丸对实验性自身免疫性葡萄膜炎大鼠NLRP3、ASC、IL-1β表达的影响 |
| 刘溪源,喻京生 |
| (湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 探讨祛风明目丸对实验性自身免疫性葡萄膜炎（experimental autoimmune uveitis, EAU）大鼠房水及视网膜组织中NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3, NLRP3）、接头蛋白凋亡相关斑点样蛋白（apoptosis related spot like protein, ASC）、白细胞介素-1β（interleukin-1β, IL-1β）表达的影响。方法 将70只SPF级雄性Lewis大鼠按随机数字表法取19只为空白组，其余为造模组，进行主动免疫，建立EAU大鼠模型，连续造模7 d后分别从造模组及空白组随机抽取3只大鼠取眼球行HE染色，进行组织病理学检查；将造模成功的EAU大鼠随机分为模型组、祛风明目丸低剂量组（简称低剂量组）和祛风明目丸正常剂量组（简称正常剂量组），每组16只。空白组、模型组予以蒸馏水灌胃，低剂量组和正常剂量组予以不同剂量的祛风明目丸药液灌胃，均干预14 d。自造模开始，每天于裂隙灯下观察眼前节变化，并采用Caspi临床分级标准评分；自灌胃起14 d后，ELISA检测房水中NLRP3、IL-1β水平，Western blot检测视网膜组织中ASC、NLRP3蛋白水平。结果 Caspi临床分级评分结果示，自造模第3天起造模组评分均明显高于空白组（P＜0.01），说明EAU模型制备成功。ELISA结果显示，与空白组相比，模型组房水中NLRP3、IL-1β水平升高（P＜0.05）；与模型组相比，正常剂量组及低剂量组两者含量均降低（P＜0.05）；正常剂量组与低剂量组差异无统计学意义（P＞0.05）。Western blot结果显示，与空白组相比，模型组视网膜组织中ASC、NLRP3蛋白表达水平升高（P＜0.05）；与模型组相比，正常剂量组及低剂量组视网膜组织中ASC、NLRP3蛋白表达含量均下降（P＜0.05）；与正常剂量组相比，低剂量组视网膜组织ASC、NLRP3蛋白表达水平升高（P＜0.05）。结论 祛风明目丸可能通过降低NLRP3炎症小体及下游炎症因子的表达对EAU大鼠发挥一定的治疗作用。 |
| 关键词: 实验性自身免疫性葡萄膜炎 祛风明目丸 NOD样受体热蛋白结构域相关蛋白3 接头蛋白凋亡相关斑点样蛋白 白细胞介素-1β |
| DOI：10.3969/j.issn.1674-070X.2023.10.005 |
| Received:January 16, 2023 |
| 基金项目:长沙市自然科学基金项目（kq2202449）。 |
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| Effects of Qufeng Mingmu Pill on NLRP3, ASC, and IL-1β expressions in rats with experimental autoimmune uveitis |
| LIU Xiyuan,YU Jingsheng |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To investigate the effects of Qufeng Mingmu Pill on the expressions of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis related spot like protein (ASC), and interleukin-1β (IL-1β) in aqueous humor and retinal tissues of rats with experimental autoimmune uveitis (EAU). Methods Nineteen of the 70 SPF-level male Lewis rats were selected as blank group according to the random number table method, the rest were made modal group for active immunization. EAU rat models were established, and 3 rats were randomly selected from the model group and the blank group after 7 days of continuous modeling to take eyeballs for HE staining and histopathological examination. The successfully modeled EAU rats were randomly divided into model group, low-dose group of Qufeng Mingmu Pill and normal-dose group of Qufeng Mingmu Pill (hereinafter referred to as the low-dose group and the normal dose group), with 16 rats in each group. The blank group and the model group were given distilled water by gavage, and the low-dose group and the normal dose group were given different doses of Qufeng Mingmu Pill liquid by gavage for 14 days. Since the beginning of molding, the changes of the anterior segment of the eye were observed under the slit lamp every day, and the Caspi clinical grading standard was used to score. After 14 days of gavage, NLRP3 and IL-1β levels in aqueous humor were checked by ELISA, and ASC and NLRP3 protein levels in retinal tissue were tested by Western blot. Results The results of Caspi clinical grading showed that the scores of the model group were significantly higher than those of the blank group since the third day of modeling (P＜0.01), indicating that the EAU model was successfully prepared. The results of ELISA showed that compared with the blank group, the NLRP3 and IL-1β levels of the model group increased (P＜0.05). Compared with the model group, the content of both normal dose group and low-dose group decreased (P＜0.05). The difference between normal dose group and low-dose group was not statistically significant (P＞0.05). The results of Western blot showed that compared with the blank group, the expression levels of ASC and NLRP3 proteins in the retinal tissues of the model group increased (P＜0.05). Compared with the model group, the expression content of ASC and NLRP3 protein in retinal tissues decreased in both normal and low-dose groups (P＜0.05). Compared with the normal dose group, the expression levels of ASC and NLRP3 protein in retinal tissue in the low-dose group were higher (P＜0.05). Conclusion Qufeng Mingmu Pill may play a therapeutic role in EAU rats by reducing the expression of NLRP3 inflammasome and downstream inflammatory factors. |
| Key words: experimental autoimmune uveitis Qufeng Mingmu Pill NOD-like receptor thermal protein domain associated protein 3 apoptosis related spot like protein interleukin-1β |
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