Tetramethylpyrazine Attenuates Oxygen-glucose Deprivation-induced Neuronal Damage through Inhibition of the HIF-1α/BNIP3 Pathway: From Network Pharmacological Finding to Experimental Validation

Shixin Xu; Nannan Zhang; Lanlan Cao; Lu Liu; Hao Deng; Shengyu Hua; Yunsha Zhang

doi:10.2174/1381612829666230215100507

Tetramethylpyrazine Attenuates Oxygen-glucose Deprivation-induced Neuronal Damage through Inhibition of the HIF-1α/BNIP3 Pathway: From Network Pharmacological Finding to Experimental Validation

Curr Pharm Des. 2023 Feb 15. doi: 10.2174/1381612829666230215100507. Online ahead of print.

Authors

Shixin Xu^{1

2}, Nannan Zhang^{1

3}, Lanlan Cao^{1

2}, Lu Liu^{1

4}, Hao Deng^{1

2}, Shengyu Hua⁵, Yunsha Zhang⁶

Affiliations

¹ Medical Experiment Center, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, China.
² Tianjin Key Laboratory of Translational Research of TCM Prescription and Syndrome, Tianjin, China.
³ Dongcheng District Community Health Service Management Center, Beijing, China.
⁴ Binhai New Area Hospital of TCM. Tian Jin, Fourth Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, China.
⁵ Institute of traditional Chinese medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
⁶ School of Integrative Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.

PMID: 36790003
DOI: 10.2174/1381612829666230215100507

Abstract

Aims: A network pharmacological analysis combined with experimental validation was used to investigate the neuroprotective mechanism of the natural product Tetramethylpyrazine(TMP).

Background: Protecting neurons is critical for acute ischemic stroke treatment. Tetramethylpyrazine is a bioactive component extracted from Chuanxiong. The neuroprotective potential of TMP has been reported, but a systematic analysis of its mechanism has not been performed.

Objective: Based on the hints of network pharmacology and bioinformatics analysis, the mechanism by which TMP alleviates oxygen-glucose deprivation-induced neuronal damage through inhibition of the HIF-1α/BNIP3 pathway was verified.

Method: In this study, we initially used network pharmacology and bioinformatics analyses to elucidate the mechanisms involved in TMP's predictive targets on a system level. The HIF-1α/BNIP3 pathway mediating the cellular response to hypoxia and apoptosis was considered worthy of focus in the bioinformatic analysis. An oxygen-glucose deprivation (OGD)-induced PC12 cell injury model was established for functional and mechanical validation. Cell viability, lactate dehydrogenase leakage, intracellular reactive oxygen species, percentage of apoptotic cells, and Caspase-3 activity were determined to assess the TMP's protective effects. Transfection with siRNA/HIF-1α or pcDNA/HIF-1α plasmids to silence or overexpress hypoxia-inducible factor 1α(HIF-1α). The role of HIF-1α in OGD-injured cells was observed first. After that, TMP's regulation of the HIF-1α/BNIP3 pathway was investigated. The pcDNA3.1/HIF-1α-positive plasmids were applied in rescue experiments.

Result: The results showed that TMP dose-dependently attenuated OGD-induced cell injury. The expression levels of HIF-1α, BNIP3, and the Bax/Bcl-2 increased significantly with increasing OGD duration. Overexpression of HIF-1α decreased cell viability, increased BNIP3 expression, and Bax/Bcl-2 ratio; siRNA-HIF-1α showed the opposite effect. TMP treatment suppressed HIF-1α, BNIP3 expression, and the Bax/Bcl-2 ratio and was reversed by HIF-1α overexpression.

Conclusion: Our study shows that TMP protects OGD-damaged PC12 cells by inhibiting the HIF-1α/BNIP3 pathway, which provides new insights into the mechanism of TMP and its neuroprotective potential.

Keywords: Apoptosis; Hypoxia-inducible factor 1α (HIF-1α); Neuroprotection; Oxygen-glucose deprivation (OGD); ROS; Tetramethylpyrazine (TMP).