Sanpian Decoction Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulating SIRT1/ERK/HIF-1α Pathway Through in Silico Analysis and Experimental Validation

Abstract

Ethnopharmacological relevance: Cerebral ischemia-reperfusion injury (CIRI) is a complex pathophysiological process involving multiple factors, and becomes the footstone of rehabilitation after ischemic stroke. Sanpian decoction (SPD) has exhibited protective effects against CIRI, migraine, and other cerebral vascular diseases. However, the underlying mechanisms have not been fully elucidated.Aim of the study: This study sought to explore the potential mechanisms underlying the effect of SPD against CIRI.

Materials and Methods: High performance liquid chromatography (HPLC) was carried out to determine the active aspects of SPD. Network pharmacology combined with experimental verification was conducted to elucidate SPD’s multi-component, multi-target, and multi-pathway mechanisms in CIRI occurrence. The pharmacodynamics of the decoction was evaluated by establishing rats model of middle cerebral artery occlusion/reperfusion (MCAO/R). In vivo and in vitro experiments, the therapeutic effect of SPD was performed using TTC, HE staining and Nissl staining. We used Tunel staining and flow cytometry to quantify cortex apoptosis. RT-qPCR and western blot were used to detect the corresponding protein and mRNA expressions, respectively.

Results: Our research showed that pretreatment with SPD improved neurological function and inhibited CIRI. Network pharmacology revealed that the hypoxia-inducible factor-1 (HIF-1) signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway-mediated apoptosis may be associated with CIRI. In vivo and in vitro experiments, we confirmed that SPD increased cerebral blood flow, improved neural function, and reduced neural apoptosis via up-regulating the expression of sirtuin 1 (SIRT1) and down-regulating phospho-extracellular regulated protein kinases (p-ERK)/ERK and HIF-1α levels in CIRI rats.

Conclusion: Taken together, the present study systematically revealed the potential targets and signaling pathways of SPD in the treatment of CIRI using in silico prediction and verified the therapeutic effects of SPD against CIRI via ameliorating apoptosis by regulating SIRT1/ERK/HIF-1α.

Note:
Funding Information: This research was supported by the National Natural Science Foundation of China (Grant No. 82174167), the key project of Hunan Province Education Department (Grant No.20A366), the project of Natural Science Foundation of Hunan Province (Grant No.2021JJ30499), the fund for Youth Top Talent Project of Hubei Provincial Health and Family Planning Commission (Grant No.EWT-2019-48), and the Postgraduate Scientific Research Innovation Project of Hunan Province (Grant No. CX20210721).

Conflict of Interests: The authors declared that they have no conflicts of interest in this work.

Ethical Approval: The National Institute of Health Guide for the Care and Use of Laboratory Animals was followed in all experimental protocols and animal-related activities. And the experiments were approved by the Laboratory Animal Ethical Committee of China Three Gorges University (No. 2020B010D).

Keywords: Sanpian decoction, Network pharmacology, cerebral ischemia-reperfusion injury, HIF-1 signaling pathway, MAPK signaling pathway, Apoptosis