Exploring the mechanisms under Zuogui Pill's treatment of ischemic stroke through network pharmacology and in vitro experimental verification

Li Li; Yan Liu; Yawei Zheng; Jian Zhu; Dan Wu; Xiaohui Yan; Changyin Li; Minghua Wu; Wenlei Li

doi:10.3389/fphar.2023.1153478

Exploring the mechanisms under Zuogui Pill's treatment of ischemic stroke through network pharmacology and in vitro experimental verification

Front Pharmacol. 2023 Jun 22:14:1153478. doi: 10.3389/fphar.2023.1153478. eCollection 2023.

Authors

Li Li^{1

2}, Yan Liu^{1

2}, Yawei Zheng^{1

2}, Jian Zhu³, Dan Wu^{1

2}, Xiaohui Yan^{1

2}, Changyin Li⁴, Minghua Wu^{1

2}, Wenlei Li^{1

2}

Affiliations

¹ Department of Neurology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.
² The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, China.
³ Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
⁴ Department of Clinical Pharmacology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.

Abstract

Due to its high mortality, incidence and disability rates, ischemic stroke poses heavy economic burdens to families and society. Zuogui Pill (ZGP) is a classic Chinese medicine for tonifying the kidney, which is effective for the recovery of neurological function after ischemic stroke. However, Zuogui Pill has not been evaluated for its potential effects on ischemic strokes. Using network pharmacology, the research aimed to explore the mechanisms of Zuogui Pill on ischemic stroke, which were further validated in SH-SY5Y cells injured by oxygen and glucose deprivation/reperfusion (OGD/R). Network analysis of Zuogui Pill identified 86 active ingredients and 107 compound-related targets correlated with ischemic stroke. Additionally, 11 core active compounds were obtained, such as Quercetin, beta sitosterol, and stigmasterol. Most of the compounds have been proven to have pharmacological activities. Based on pathway enrichment studies, Zuogui Pill may exert neuroprotection through MAPK signaling, PI3K-Akt signaling and apoptosis, as well as enhance neurite outgrowth and axonal regeneration effect via mTOR signaling, p53 signaling and Wnt signaling pathways. In vitro experiment, the viability of ischemic neuron treated with Zuogui Pill was increased, and the ability of neurite outgrowth was significantly improved. Western blot assays shown that the pro-neurite outgrowth effect of Zuogui Pill on ischemic stroke may be relate to PTEN/mTOR signal pathway. The results of the study provided new insights into Zuogui Pill's molecular mechanism in treatment of ischemic stroke, as well as clinical references for its use.

Keywords: SH-SY5Y cells; ischemic stroke; network pharmacology; neurite outgrowth; zuogui pill.

Grants and funding

This work was supported by National Natural Science Foundation of China (Grant Nos. 81973794), Jiangsu Province Administration of Chinese Medicine (Grant No. JD201803), National Natural Science Foundation of China (Grant No. 82274428), Jiangsu Province High level Health Talents “Six One Project” Top Talent Project (Grant No. LGY201806), National Administration of Traditional Chines Medicine: Evidence-Based Capacity Building Project (Grant No. 2019XZZX-NB007), 333 high level talentstraining project in Jiangsu (Grant No. BRA2016507), Jiangsu Province Administration of Chinese Medicine (Grant No. ZT202102).