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黄云,陈姝,夏如梅,李蕾,喻干龙,喻娟.基于网络药理学和实验验证探讨近视复明丸治疗近视的作用机制[J].湖南中医药大学学报英文版,2023,43(12):2223-2230.[Click to copy
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| 基于网络药理学和实验验证探讨近视复明丸治疗近视的作用机制 |
| 黄云,陈姝,夏如梅,李蕾,喻干龙,喻娟 |
| (湖南中医药大学, 湖南 长沙 410208;中医药防治眼耳鼻咽喉疾病湖南省重点实验室, 湖南 长沙 410208;湖南中医药大学第一中医临床学院, 湖南 长沙 410007) |
| 摘要: |
| 目的 探讨近视复明丸治疗近视的作用机制。方法 检索TCMSP、TCMID数据库获取近视复明丸中药有效成分，检索SwissTargetPrediction数据库获取有效成分的靶点信息，检索GeneCards、DrugBank、OMIM、Therapeutic Target Database获取近视的主要靶点；运用Cytoscape3.9.1软件和STRING平台构建蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络图；通过DAVID数据库进行GO功能和KEGG通路富集分析。选取48只3周龄雄性普通级豚鼠，分为空白组、近视模型组、中药高剂量组、中药低剂量组。模型复制成功后给予药物干预。予近视模型组2 mL生理盐水灌胃；中药高剂量组予2 mL近视复明丸药粉（给药浓度为0.8 g/mL）灌胃；中药低剂量组予2 mL近视复明丸药粉（给药浓度为0.2 g/mL）灌胃。空白组予2 mL生理盐水灌胃。每日1次，共4周。HE染色观察巩膜组织病理变化，RT-qPCR检测巩膜中基质金属蛋白酶-2（matrix metalloproteinase-2，MMP-2）和基质金属蛋白酶抑制剂-2（tissue inhibitor of matrix metalloproteinase-2，TIMP-2）表达水平。结果 近视复明丸包括有效成分587个，相关作用靶点1 760个，药物疾病的共同靶点122个。网络分析的结果获得丝氨酸-苏氨酸激酶（serine-threonine kinase 1，AKT1）、血管内皮生长因子A（vascular endothelial growth factor A，VEGFA）、白细胞介素-1β（interlukin-1β，IL-1β）、基质金属蛋白酶抑制剂-9（matrix metalloproteinase-9，MMP-9）、表皮生长因子受体（epidermal growth factor receptor，EGFR）、MMP-2等关键靶点。GO功能富集分析得到631个条目，KEGG通路富集筛选出151条信号通路。动物实验结果显示：近视复明丸能够明显降低豚鼠巩膜组织MMP-2mRNA表达水平（P<0.01），同时提高TIMP-2 mRNA表达水平（P<0.05），促进巩膜胶原纤维合成，减少巩膜胶原纤维降解。结论 近视复明丸能降低MMP-2 mRNA表达水平，升高TIMP-2 mRNA表达水平，进而调控巩膜重塑，延缓近视的进展。 |
| 关键词: 近视复明丸 网络药理学 近视 巩膜 基质金属蛋白酶-2 豚鼠 |
| DOI：10.3969/j.issn.1674-070X.2023.12.012 |
| Received:May 19, 2023 |
| 基金项目:国家自然科学基金项目（81603665）；湖南省中医药管理局课题（2021057）；湖南省杰出创新青年项目（kq2107014）；湖南省研究生科研创新课题（CX20220813）。 |
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| Mechanism of action of Jinshi Fuming Pill in treating myopia based on network pharmacology and experimental verification |
| HUANG Yun,CHEN Shu,XIA Rumei,LI Lei,YU Ganlong,YU Juan |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Key Laboratory for Prevention & Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine, Changsha, Hunan 410208, China;The First Clinical School of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To explore the mechanism of action of Jinshi Fuming Pill(JSFMP) in treating myopia.Methods TCMSP and TCMID databases were searched for the active ingredients of JSFMP, SwissTargetPrediction database for the target information of active ingredients, and GeneCards, DrugBank, OMIM, and Therapeutic Target Database for the main targets of myopia; Cytoscape3.9.1 software and STRING platform were used to construct protein-protein interaction networks(PPI); GO function and KEGG pathway enrichment analysis were performed through DAVID database. Forty-eight three-week-old male guinea pigs were selected and divided into blank group, myopia model group, high-and low-dose JSFMP groups, and drug intervention was given after successful model replication. Myopic model group was given 2 mL normal saline by gavage, the high-dose JSFMP group was given2 mL JSFMP powder(0.8 g/mL) by intragastric administration, the low-dose JSFMP group was given 2 mL JSFMP powder(0.2 g/m L)by gavage, and the blank group was given 2 mL normal saline by intragastric administration, respectively, once a day for 4 weeks.HE staining was used to observe the pathological changes of sclera, and RT-qPCR was used to examine the expression levels of matrix metallo proteinase-2(MMP-2) and tissue inhibitor of matrix metalloproteinase-2(TIMP-2) in sclera.Results There were 587active ingredients, 1 760 related targets and 122 common targets of drug and disease in JSFMP. Network analysis obtained some key targets such as serine-threonine kinase 1(AKT1), vascular endothelial growth factor A(VEGFA), interleukin-1β(IL-1β), matrix metalloproteinase-9(MMP-9), epidermal growth factor receptor(EGFR), and MMP-2. GO functional enrichment analysis yielded 631items, and KEGG pathway enrichment screened out 151 signaling pathways. Animal experiments showed that JSFMP could significantly reduce MMP-2 mRNA expression level in sclera tissues of guinea pigs(P<0.01), increase TIMP-2 mRNA expression level(P<0.05), promote the synthesis of scleral collagen fibers, and reduce the degradation of scleral collagen fibers.Conclusion JSFMP can decrease MMP-2 mRNA expression level and increase TIMP-2 mRNA expression level in sclera tissues, thereby regulating scleral remodeling and delay the progression of myopia. |
| Key words: Jinshi Fuming Pill network pharmacology myopia sclera matrix metalloproteinase-2 guinea pig |
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