LC-MS/MS Tracks Herbal Stroke Remedy in the Brain

The traditional Chinese medicine (TCM) Suhexiang Wan is a herbal remedy that has long been used to treat stroke-related problems, such as sudden loss of consciousness, trouble with brain and nerve function, and paralysis on one side of the body. Modern studies suggest it may protect the brain and reduce inflammation. However, it is still unclear which of its ingredients reach the brain, how they might help in ischemic stroke (IS), a stroke caused by a blocked blood vessel, and how they work at the molecular level.

To shed light on this, researchers set out to identify which of the remedy’s ingredients make their way into the brain and to study one promising ingredient that might help protect against the damage that can occur when blood flow returns to the brain after a stroke. They paid particular attention to the blood-brain barrier, the protective layer that controls what passes from the blood into the brain. Using animals in which a major blood vessel in the brain was temporarily blocked and then reopened to mimic a stroke, they analyzed brain tissue with ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The team published their findings in the Journal of Ethnopharmacology.¹

What Is Suhexiang Wan, and Why Are Researchers Studying It for Stroke?

Suhexiang Wan is a classic example of a group of fragrant remedies used in traditional Chinese medicine to help revive people who have lost consciousness. It has long been used to treat stroke-related problems, such as sudden fainting, trouble with brain and nerve function, and paralysis on one side of the body.² This long history of use gives researchers good reason to study how the remedy might protect against ischemic stroke and how it works.³

How Was the Study Carried Out?

The researchers used computer-based analysis to predict which parts of the body the remedy’s ingredients might act on and which biological processes they might affect. One ingredient, piperlongumine, a natural compound from long pepper, was then confirmed and measured in both the remedy and brain tissue by comparing it with a pure reference sample using liquid chromatography-tandem mass spectrometry (LC-MS/MS). To see whether piperlongumine interacts with STAT3, a protein that helps switch on inflammation, the team used computer simulations along with laboratory tests on purified proteins and cells. They tested the protective effects in mice with a simulated stroke and in brain blood vessel cells that were deprived of oxygen and sugar and then given them back, mimicking what happens during a stroke and recovery.¹

Of the 1991 ingredients tentatively identified in the remedy, 55 were found in the brains of the mice with simulated strokes. In these mice, the remedy greatly improved brain and nerve function, reduced the amount of brain tissue damaged and the swelling in the brain, and made the blood-brain barrier less leaky. It also restored the proteins that seal the gaps between the cells lining the brain’s blood vessels.¹

What Did the Study Reveal About How Suhexiang Wan and Piperlongumine Protect the Brain After a Stroke?

Piperlongumine was chosen for closer study because it reached the brain, appeared to play a central role in the predicted network of effects, was expected to interact with STAT3, and had previously been linked to protection against this kind of injury. The remedy contained about 48 micrograms of piperlongumine per gram, and two hours after the mice were given the remedy, their brains contained about 160 nanograms per gram. Laboratory tests supported the idea that piperlongumine binds to STAT3.¹

On its own, piperlongumine reproduced some of the remedy’s protective effects. It improved brain and nerve function, helped keep the blood-brain barrier intact, restored the barrier function of the blood vessel cells, and reduced the inflammation linked to STAT3. Further experiments supported STAT3’s role. Lowering STAT3 levels in the cells partly mimicked piperlongumine’s protective effects, while restoring STAT3 undid that protection. When STAT3 was restored, piperlongumine still reduced the damage, suggesting that it protects the blood-brain barrier at least partly by acting on STAT3.¹

This study,” write the authors of the paper,¹ “provides new insights into the pharmacological basis of SHXW in IS by integrating chemical profiling, brain distribution analysis, and pharmacological validation. These results,” they continue, “provide pharmacological evidence linking the traditional application of SHXW with its modern molecular mechanisms while highlighting brain-distribution-guided approaches for investigating the therapeutic basis of complex TCM formulas.”¹

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References

1. Wang, J.; Ma, S.; Yu, D. et al. Pharmacological Basis of Suhexiang Wan in Ischemic Stroke: Identification and Functional Characterization of Piperlongumine as a Brain-Distributed Bioactive Constituent Associated with STAT3-mediated Blood-Brain Barrier Protection. J Ethnopharmacol. 2026, 122561. DOI: 10.1016/j.jep.2026.122561
2. Zhou, M.; Li, D.; Li, L. et al. Post-stroke Treatment of Storax Improves Long-term Outcomes of Stroke in Rats. J Ethnopharmacol. 2021, 280, 114467. DOI: 10.1016/j.jep.2021.114467
3. Lai, X.; Xiong, X.; Jia, Q. et al. Suhexiang Pill for Acute Ischemic Stroke in Real-world Practice Setting (SUNRISE): Protocol of a Multicenter Registry. BMC Complement Med Ther. 2025, 25 (1), 30. DOI: 10.1186/s12906-025-04762-9