Title:Tetramethylpyrazine Attenuates Oxygen-glucose Deprivation-induced Neuronal
Damage through Inhibition of the HIF-1α/BNIP3 Pathway: from Network
Pharmacological Finding to Experimental Validation
Volume: 29
Issue: 7
Author(s): Shixin Xu*, Nannan Zhang, Lanlan Cao, Lu Liu, Hao Deng, Shengyu Hua and Yunsha Zhang
Affiliation:
- Medical Experiment Center, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, China
- Tianjin
Key Laboratory of Translational Research of TCM Prescription and Syndrome, Tianjin, China
Keywords:
Tetramethylpyrazine, neuroprotection, oxygen-glucose deprivation, hypoxia-inducible factor 1α, apoptosis, ROS.
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.
Methods: 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.
Results: 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.