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曾令聪, 吴凯, 朱冰瑶, 刘倩宏, 陈雄, 彭清华, 姚小磊.除风益损汤对兔角膜损伤后自噬相关因子及TGF-β1、MMP-2表达的影响[J].湖南中医药大学学报英文版,2025,45(2):212-219.[Click to copy
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| 除风益损汤对兔角膜损伤后自噬相关因子及TGF-β1、MMP-2表达的影响 |
| 曾令聪,吴凯,朱冰瑶,刘倩宏,陈雄,彭清华,姚小磊 |
| (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;郴州市中医医院, 湖南 郴州 424499;深圳市中西医结合医院, 广东 深圳 518033) |
| 摘要: |
| 目的 探讨除风益损汤对兔角膜损伤的修复作用及对自噬相关因子和转化生长因子-β1（TGF-β1）、基质金属蛋白酶-2（MMP-2）的影响。方法 随机将84只新西兰大白兔分为6组，具体为空白组、模型组、中药组、自噬促进组、自噬抑制组、自噬促进+中药组、自噬抑制+中药组，每组12只，每一组下又分为3个亚组，每个亚组4只，分别供造模后第7、14、28天检测。动物模型建立后各组在第7、14、28天观察角膜以及角膜荧光素染色情况，记录角膜混浊评分，采用激光共焦断层扫描仪观察角膜基质层修复情况，用Western blot检测角膜中TGF-β1、微管相关蛋白轻链3（LC3）比值、MMP-2的蛋白表达。结果 （1）角膜基质激光共焦断层扫描结果：除空白组外，其余各组角膜在造模后第7天均有成纤维细胞浸润及胶原沉积。随着时间的推移，中药组、自噬促进组、自噬促进+中药组中成纤维细胞及胶原沉积较其他组减少。（2）显微镜观察白兔角膜情况：空白组白兔的角膜在第7、14、28天皆清晰透明。其余各组白兔的角膜在第7天均出现混浊，荧光素着染，随着时间推移荧光素着色范围缩小，角膜混浊逐渐消褪。角膜混浊评分显示，与模型组相比，第28天自噬抑制组、自噬抑制+中药组评分较高（P<0.05）。（3）LC3-Ⅱ/Ⅰ：与模型组相比，各时间段中药组、自噬促进组LC3-Ⅱ/Ⅰ表达均升高（P<0.05），自噬抑制组表达降低（P<0.05）。（4）TGF-β1：与模型组相比，各时间段中药组、自噬促进组表达均较低（P<0.05），自噬抑制组表达较高（P<0.05）。（5）MMP-2：与模型组相比，各时间段中药组、自噬促进组表达均较低（P<0.05），自噬抑制组表达较高（P<0.05）。结论 除风益损汤、氯化锂可以促进角膜损伤的愈合，有助于减少瘢痕形成，其机制是通过促进自噬和调控TGF-β1、MMP-2的水平来实现的。 |
| 关键词: 除风益损汤 角膜损伤 自噬 微管相关蛋白轻链3 转化生长因子-β1 基质金属蛋白酶-2 |
| DOI：10.3969/j.issn.1674-070X.2025.02.004 |
| Received:September 02, 2024 |
| 基金项目:国家自然科学基金面上项目（82174443）；湖南省教育厅“芙蓉学者”奖励计划（2022年）；湖南省卫生健康委高层次人才计划（2023年）；湖南省眼科疾病（中医）临床医学研究中心（2023SK4038）；湖南中医药大学一流学科开放基金（2021ZYX05）。 |
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| Effects of Chufeng Yisun Decoction on autophagy related factors and expressions of TGF-β1 and MMP-2 after corneal injury in rabbits |
| ZENG Lingcong, WU Kai, ZHU Bingyao, LIU Qianhong, CHEN Xiong, PENG Qinghua, YAO Xiaolei |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Chenzhou Municipal Hospital of Traditional Chinese Medicine, Chenzhou, Hunan 424499, China;Shenzhen Hospital of Integrated Chinese and Western Medicine, Shenzhen, Guangdong 518033, China) |
| Abstract: |
| Objective To investigate the repair effects of Chufeng Yisun Decoction on corneal injury in rabbits and its impact on autophagy related factors, as well as the expressions of TGF-β1, and MMP-2. Methods A total of 84 New Zealand white rabbits were randomized into six groups, including blank group, model group, Chinese medicines group, autophagy promotion group, autophagy inhibition group, autophagy promotion+Chinese medicines group and autophagy inhibition+Chinese medicines group, with 12 rabbits in each group. Each group was further divided into three subgroups of four rabbits each, for observations and testing at 7, 14, and 28 days post-modeling. After establishing the animal model, the corneal condition and corneal fluorescein staining were observed on days 7, 14, and 28 in each group, and the corneal opacity score was recorded. A confocal laser scanning microscopy was used to observe the repair of the corneal stroma. Western blot was employed to determine the expressions of transforming growth factor-β1 (TGF-β1), the ratio of microtubule-associated protein light chain 3 (LC3)-II/I, and matrix metalloproteinase-2 (MMP-2) in the cornea. Results (1) Confocal laser scanning of the corneal stroma: Except for the blank group, all other groups showed fibroblast infiltration and collagen deposition in the cornea on day 7 post-modeling. Over time, the Chinese medicines group, autophagy promotion group, and autophagy promotion+Chinese medicines group exhibited reduced fibroblast and collagen deposition compared to the other groups; (2) Microscopic observation of rabbit corneas: The corneas of rabbits in the blank group were clear and transparent on days 7, 14, and 28. The corneas of rabbits in all other groups were hazy and stained with fluorescein on day 7, with the fluorescein staining area decreasing and corneal opacity gradually fading over time. Corneal opacity score showed that, compared with the model group, the autophagy inhibition and autophagy inhibition+Chinese medicines groups had higher scores on day 28 (P<0.05). (3) LC3-Ⅱ/Ⅰ: Compared with the model group, the LC3-Ⅱ/Ⅰ expression was higher in the Chinese medicines and autophagy promotion groups at all time points (P<0.05), while it was lower in the autophagy inhibition group (P<0.05). (4) TGF-β1: Compared with the model group, the TGF-β1 expression was lower in the Chinese medicines and autophagy promotion groups at all time points (P<0.05), while it was higher in the autophagy inhibition group (P<0.05). (5) MMP-2: Compared with the model group, the MMP-2 expression was lower in the Chinese medicines and autophagy promotion groups at all time points (P<0.05), while it was higher in the autophagy inhibition group (P<0.05). Conclusion Chufeng Yisun Decoction and lithium chloride can promote the healing of corneal injuries and help reduce scar formation, and their mechanisms involve promoting autophagy and regulating the levels of TGF-β1 and MMP-2. |
| Key words: Chufeng Yisun Decoction corneal injury autophagy microtubule-associated protein light chain 3 transforming growth factor-β1 matrix metalloproteinase-2 |
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