# Single-cell RNA-seq reveals the piperlongumine is a potential drug for ischemic stroke

**Authors:** Jinwei Li, Xiaosi Zhu, Yan Zhang, Zixia Xu, Yurui Zhuang, Liqi Chen, Yan Cao, Chuan Chang, Yao Lv

PMC · DOI: 10.1371/journal.pone.0340725 · PLOS One · 2026-01-23

## TL;DR

This study uses single-cell RNA sequencing to identify piperlongumine as a potential treatment for ischemic stroke by analyzing molecular pathways and drug interactions.

## Contribution

The study proposes piperlongumine as a novel therapeutic candidate for ischemic stroke through computational and experimental validation.

## Key findings

- Apoptosis is significantly increased after ischemic brain injury.
- Piperlongumine binds to proteins encoded by Actb and Cflar, suggesting therapeutic potential.
- In vitro and in vivo experiments confirmed the effectiveness of piperlongumine.

## Abstract

Ischemic stroke is a cerebrovascular disease that can cause long-term neurological impairment, dementia, or death. It is the third most common cause of disability and the second leading cause of death worldwide. The aim of this study was to explore the underlying molecular mechanisms and potentially effective therapeutic drugs for ischemic stroke. Single-cell seq data (GSE174574) were downloaded from the Gene Expression Omnibus (GEO) database, and dimensionality reduction clustering was performed after quality control. Eighteen cell clusters were identified, which were annotated into 9 cell types according to specific marker genes. In addition, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis (GSEA) enrichment analysis showed that apoptosis was significantly increased after ischemic brain injury, while p53 signaling pathway, TNF-a signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway may play an important role in ischemic stroke. Furthermore, Connectivity Map (CMap) analysis and molecular docking suggested that piperlongumine might be an effective drug for the treatment of ischemic stroke by binding to the proteins encoded by Actb and Cflar. Finally, in vitro and in vivo experiments conformed the effectiveness of piperlongumine. This study provided new ideas for the treatment of ischemic stroke.

## Linked entities

- **Genes:** ACTB (actin beta) [NCBI Gene 60], CFLAR (CASP8 and FADD like apoptosis regulator) [NCBI Gene 8837]
- **Chemicals:** piperlongumine (PubChem CID 637858)
- **Diseases:** ischemic stroke (MONDO:1060198)

## Full-text entities

- **Genes:** S100A8 (S100 calcium binding protein A8) [NCBI Gene 6279] {aka 60B8AG, CAGA, CFAG, CGLA, CP-10, L1Ag}, CAMP (cathelicidin antimicrobial peptide) [NCBI Gene 820] {aka CAP-18, CAP18, CRAMP, FALL-39, FALL39, HSD26}, Rspo3 (R-spondin 3) [NCBI Gene 72780] {aka 2810459H04Rik, Cristin1, Thsd2}, Bax (BCL2-associated X protein) [NCBI Gene 12028], Cflar (CASP8 and FADD-like apoptosis regulator) [NCBI Gene 12633] {aka 2310024N18Rik, A430105C05Rik, CLARP, Cash, Casper, FLAME}, Rbfox3 (RNA binding protein, fox-1 homolog (C. elegans) 3) [NCBI Gene 52897] {aka Fox-3, Hrnbp3, NeuN, Neuna60}, Tjp1 (tight junction protein 1) [NCBI Gene 292994] {aka ZO-1}, HEXB (hexosaminidase subunit beta) [NCBI Gene 3074] {aka ENC-1AS, HEL-248, HEL-S-111}, CFLAR (CASP8 and FADD like apoptosis regulator) [NCBI Gene 8837] {aka CASH, CASP8AP1, CLARP, Casper, FLAME, FLAME-1}, S1pr1 (sphingosine-1-phosphate receptor 1) [NCBI Gene 29733] {aka Edg1}, Hamp (hepcidin antimicrobial peptide) [NCBI Gene 84604] {aka Hepc}, PLP1 (proteolipid protein 1) [NCBI Gene 5354] {aka GPM6C, HLD1, MMPL, PLP, PLP/DM20, PMD}, ACTA2 (actin alpha 2, smooth muscle) [NCBI Gene 59] {aka ACTSA, SMDYS}, Mapk1 (mitogen-activated protein kinase 1) [NCBI Gene 26413] {aka 9030612K14Rik, ERK, Erk2, MAPK2, PRKM2, Prkm1}, Cldn5 (claudin 5) [NCBI Gene 65131], IGFBP2 (insulin like growth factor binding protein 2) [NCBI Gene 3485] {aka IBP2, IGF-BP53}, Ctsl (cathepsin L) [NCBI Gene 13039] {aka 1190035F06Rik, CatL, Ctsl1, MEP, fs, nkt}, JUN (Jun proto-oncogene, AP-1 transcription factor subunit) [NCBI Gene 3725] {aka AP-1, AP1, c-Jun, cJUN, p39}, ACTB (actin beta) [NCBI Gene 60] {aka BKRNS, BNS, BRWS1, CSMH, DDS1, PS1TP5BP1}, Tnfrsf1a (tumor necrosis factor receptor superfamily, member 1a) [NCBI Gene 21937] {aka CD120a, FPF, TNF-R, TNF-R-I, TNF-R1, TNF-R55}, Mapk8 (mitogen-activated protein kinase 8) [NCBI Gene 26419] {aka JNK, JNK1, Prkm8, SAPK1}, Trp53-ps (transformation related protein 53, pseudogene) [NCBI Gene 22060], Cd4 (Cd4 molecule) [NCBI Gene 24932] {aka W3/25, p55}, TP53 (tumor protein p53) [NCBI Gene 7157] {aka BCC7, BMFS5, LFS1, P53, TRP53}, MAPT (microtubule associated protein tau) [NCBI Gene 4137] {aka DDPAC, FTD1, FTDP-17, MAPTL, MSTD, MTBT1}, TNF (tumor necrosis factor) [NCBI Gene 7124] {aka DIF, IMD127, TNF-alpha, TNFA, TNFSF2, TNLG1F}, Tnf (tumor necrosis factor) [NCBI Gene 21926] {aka DIF, TNF-a, TNF-alpha, TNFSF2, TNFalpha, Tnfa}, Ctsb (cathepsin B) [NCBI Gene 13030] {aka APPM, CB}, Casp8 (caspase 8) [NCBI Gene 12370] {aka CASP-8, FLICE, MACH, Mch5}, CCL5 (C-C motif chemokine ligand 5) [NCBI Gene 6352] {aka D17S136E, RANTES, SCYA5, SIS-delta, SISd, TCP228}, S100A9 (S100 calcium binding protein A9) [NCBI Gene 6280] {aka 60B8AG, CAGB, CFAG, CGLB, L1AG, LIAG}, Ctsd (cathepsin D) [NCBI Gene 13033] {aka CD, CatD}, Jun (Jun proto-oncogene, AP-1 transcription factor subunit) [NCBI Gene 16476] {aka AP-1, Junc, c-jun}, Atf2 (activating transcription factor 2) [NCBI Gene 11909] {aka Atf-2, CRE-BP, Creb2, D130078H02Rik, Tg(Gzma-Klra1)7Wum, mXBP}, Actb (actin, beta) [NCBI Gene 11461] {aka Actx, E430023M04Rik, beta-actin}, Slc40a1 (solute carrier family 40 member 1) [NCBI Gene 170840] {aka Fpn1, Slc11a3, Slc39a1}
- **Diseases:** hemorrhagic (MESH:D006470), brain parenchymal damage (MESH:D002543), neural damage (MESH:D015441), dementia (MESH:D003704), Foot (MESH:D005530), neuronal loss (MESH:D009410), neurological impairment (MESH:D009422), neuroinflammation (MESH:D000090862), cerebrovascular disease (MESH:D002561), brain damage (MESH:D001925), Ischemic stroke (MESH:D002544), Cerebral ischemia (MESH:D002545), stroke (MESH:D020521), inflammation (MESH:D007249), dislocation (MESH:D004204), Brain injury (MESH:D001930), cerebral ischemic injury (MESH:D017202), necrotic (MESH:D009336), pain (MESH:D010146), nerve damage (MESH:D000080902), death (MESH:D003643), vasogenic edema (MESH:D001929), cerebral I/R injury (MESH:C580424), neurological and mental damage (MESH:D020196), amyloid (MESH:C000718787), atherosclerosis (MESH:D050197), MCAO (MESH:D020244), neurological deficits (MESH:D009461)
- **Chemicals:** iron (MESH:D007501), cobalt 60 (MESH:C000615395), ASP-105 (-), Piperlongumine (MESH:C498077), amino acid (MESH:D000596), DAPI (MESH:C007293), glucose (MESH:D005947), Alexa Fluor 488 (MESH:C000711379), water (MESH:D014867), isoflurane (MESH:D007530), alkaloid (MESH:D000470), CCK-8 (MESH:D012844), gemcitabine (MESH:D000093542), CO2 (MESH:D002245), oxygen (MESH:D010100), streptomycin (MESH:D013307), PI (MESH:D011419), Triton X-100 (MESH:D017830), penicillin (MESH:D010406)
- **Species:** Piper longum (species) [taxon 49511], Homo sapiens (human, species) [taxon 9606], Mus musculus (house mouse, species) [taxon 10090], Rattus norvegicus (brown rat, species) [taxon 10116]
- **Mutations:** C1062S, A-to-I
- **Cell lines:** bEnd.3 — Mus musculus (Mouse), Transformed cell line (CVCL_0170)

## Full text

_Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12829879/full.md

## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12829879/full.md

## References

40 references — full list in the complete paper: https://tomesphere.com/paper/PMC12829879/full.md

---
Source: https://tomesphere.com/paper/PMC12829879