Mengmeng Yang,1,* Hualin Liu,1,* Jiewen Zhou,2,* Kewei Wang,1 Yujing Sun,1 Na Ning,3 Qiuling Huang,3 Jiajia Hu,3,4 Jidong Liu,1,5– 7 Fei Yan,1,5– 7 Xinguo Hou,1,5– 7 Li Chen,1,5– 7 Lingshu Wang,1,5– 7 Fuqiang Liu1,5– 7 

1Department of Endocrinology, Qilu Hospital of Shandong University, Jinan, Shandong, 250000, People’s Republic of China; 2The First Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510000, People’s Republic of China; 3Guangzhou Baiyunshan Zhongyi Pharmaceutical Co., Ltd, Guangzhou, Guangdong, 510000, People’s Republic of China; 4Wenzhou Medical University, Wenzhou, Zhejiang, 325000, People’s Republic of China; 5Institute of Endocrine and Metabolic Diseases of Shandong University, Jinan, Shandong, 250000, People’s Republic of China; 6Key Laboratory of Endocrine and Metabolic Diseases, Shandong Province Medicine & Health, Jinan, Shandong, 250000, People’s Republic of China; 7Jinan Clinical Research Center for Endocrine and Metabolic Disease, Jinan, Shandong, 250000, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Fuqiang Liu, Department of Endocrinology, Qilu Hospital of Shandong University, Jinan, Shandong, 250000, People’s Republic of China, Email liufuqiang@sdu.edu.cn Lingshu Wang, Department of Endocrinology, Qilu Hospital of Shandong University, Jinan, Shandong, 250000, People’s Republic of China, Email wanglingshu@qiluhospital.com

Background: Zhangyanming Tablets (ZYMT) is a proprietary Chinese medicine containing a variety of traditional Chinese medicines, which can be used to treat a wide range of eye diseases, but its exact effect on diabetic retinopathy (DR) and the specific mechanism are still unclear. This study aims to investigate the ameliorative effects and specific mechanisms of ZYMT on DR.
Methods: Key regulatory genes and potential therapeutic targets of ZYMT for DR were evaluated using network pharmacological analysis. Diabetic db/db mice were given low-dose ZYMT (330 mg/kg) and high-dose ZYMT (660 mg/kg), and relevant metabolic indices were tested. Histochemical staining and optical coherence tomography angiography (OCTA) were used to evaluate the histopathological structure of mice retina, RT-qPCR, TUNEL staining and immunofluorescence staining were used to evaluate the anti-apoptosis and anti-angiogenesis effect of ZYMT on DR.
Results: The results of network topology analysis showed that the top 10 Traditional Chinese Medicine (TCM) ingredients of ZYMT were quercetin, luteolin, kaempferol, wogonin, naringenin, β-sitosterol, baicalein, isorhamnetin, acacetin, and stigmasterol. ZYMT treats DR through key nodes such as AKT1, TNF, MAPK8, RELA, VEGFA, HIF1A, IL6, CASP3, BCL2, STAT3, and ICAM1. ZYMT has a direct effect on DR rather than secondary improvement of metabolic indices. Tissue staining demonstrated that ZYMT improved retinal vascular morphology and delayed retinal thinning in db/db mice. The OCTA imaging also showed that ZYMT increased blood flow density in db/db mice. TUNEL staining and RT-qPCR results showed that ZYMT could reduce the apoptosis of retinal cells in db/db mice, and RT-qPCR and immunofluorescence staining showed that ZYMT could inhibit retinal neovascularization.
Conclusion: This study found the potential target of ZYMT to ameliorate DR through network pharmacological analysis, and verified that ZYMT can improve DR by exerting anti-apoptosis and anti-neovascularization.

Keywords: diabetic retinopathy, zhangyanming tablets, network pharmacological analysis, apoptosis, neovascularization