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PM · Lens · Investigating the Protective Mechanism of the Zha-Chong-Shi-San-Pill Ethanol Extract on Cerebral Stroke Based on Network Pharmacology, Molecular Docking, and in vitro Experiments

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Article · Volume 21, Issue 4 · pp. 1597–1607

Investigating the Protective Mechanism of the Zha-Chong-Shi-San-Pill Ethanol Extract on Cerebral Stroke Based on Network Pharmacology, Molecular Docking, and in vitro Experiments

Hua Jin1, Ta Na1, Yue-hui Liu2,3, Qian-qian Ma3, Ming-qi Liu2, Ruo-shui Zhou1, Liang Wei1, Hong-quan Jin4, Dong-wei Zhang1

  1. 1Affiliated Hospital of Inner Mongolian Minzu University, Tongliao, Inner Mongolia, CHINA.
  2. 2Department of Neurology, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, CHINA.
  3. 3Inner Mongolia Minzu University, Tongliao, Inner Mongolia, CHINA.
  4. 4Department of Neurology, The Beidahuang Group Beian Hospital, Heihe, Heilongjiang, CHINA.

Abstract

Objectives: To study the antioxidant effects of the traditional Mongolian medicine Zha-Chong-Shi-San-Pill ethanol extract (EEZC) on cerebral stroke for future clinical use.Materials and Methods: Active components and mechanisms of EEZC against cerebral stroke were predicted and analyzed via ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS), network pharmacology, and molecular docking. Next, an oxidative damage model was established in bEnd.3 cells using H2O2 to simulate cerebral stroke damage. Multiple cellular tests were performed to verify the mechanisms, including cell counting kit-8 (CCK-8), flow cytometry, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining, enzymelinked immunosorbent assay (ELISA), and Western blotting.Results: It was found that EEZC conferred protection against cerebral stroke H2O2-treated bEnd.3 cells. Moreover, in vitro experiments revealed that EEZC increased cell survival rates and suppressed cell apoptosis and reactive oxygen species (ROS) release. Moreover, EEZC reduced the levels of interleukin-6 (IL-6) and MMP-9, and increased the levels of interleukin-10 (IL-10), superoxide dismutase (SOD), glutathione (GSH), vascular endothelial growth factor (VEGF), phospho-endothelial nitric oxide synthase (p-eNOS), phospho-phosphatidylinositide 3-kinases (p-pi3k), and phospho-protein kinase B (p-AKT).Conclusion: These results strongly suggested that EEZC might protect against cerebral stroke via the PI3K/Akt/eNOS signaling pathway.

Keywords: Zha-Chong-Shi-San-Pill ethanol extract (EEZC); Pi3K/AKT/eNOS pathway; network pharmacology; cerebral stroke

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