αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke
IntroductionαAsarone, an essential oil derived from Acorus gramineus Aiton, which has been successfully used to treat epilepsy in traditional chinese medicine, and has also been reported to confer neuroprotective effects on stroke. However, its mechanism of action remains poorly understood.MethodsTh...
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Frontiers Media S.A.
2025-01-01
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2025.1504683/full |
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| author | Yonghuan Yan Linfang Wu Lu Wang Dandan Wang Mengting Huang Jinyong Peng Jinyong Peng Yingying Huang Yingying Huang Yingying Huang |
| author_facet | Yonghuan Yan Linfang Wu Lu Wang Dandan Wang Mengting Huang Jinyong Peng Jinyong Peng Yingying Huang Yingying Huang Yingying Huang |
| author_sort | Yonghuan Yan |
| collection | DOAJ |
| description | IntroductionαAsarone, an essential oil derived from Acorus gramineus Aiton, which has been successfully used to treat epilepsy in traditional chinese medicine, and has also been reported to confer neuroprotective effects on stroke. However, its mechanism of action remains poorly understood.MethodsThe effects of αAsarone on autophagy were examined by WB, RT-qPCR, immunofluorescence colocalization, transmission electron microscope, and autophagic flux activity was measured by infecting HT22 cells with mRFP-GFP-LC3 adenovirus. And then, cells were transfected with both mimic-miR-499-5p and inhibit-miR-499-5p to investigate the role of miR-499-5p in regulating the effects of αAsarone on stroke. To further clarify the protective effect of αAsarone in vivo, TTC staining, neurological function score, H&E staining, Nissl staining, Laser speckle contrast imaging, transmission electron microscopy, immunofluorescence colocalization, WB and RT-qPCR were performed in the MCAO mice.ResultsαAsarone was observed to inhibit the apoptosis of neuronal cells, and enhance autophagy. In addition, αAsarone promoted the expression of miR-499-5p. Targeting miR-499-5p can negatively regulate PDCD4 expression and the results from the dual-luciferase reporter assay demonstrate the direct targeting of PDCD4 by miR-499-5p. Promoting miR-499-5p can decrease the expression of PDCD4, increase ATG5, and enhance the protective effect of αAsarone on OGD/R injury while inhibiting miR-499-5p can weaken the effect of αAsarone. In vivo experiments further confirmed that αAsarone improved mice MCAO as evidenced by the amelioration of the neurological deficits and facilitated neuronal autophagy. Furthermore, we found that αAsarone reversed the effect of chloroquine, an autophagy inhibitor, and enhanced neuronal autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway.DiscussionOur data suggest that αAsarone alleviates neuronal injury of stroke by facilitating neuronal autophagy through the miR-499-5p/PDCD4/ATG5 signaling pathway. |
| format | Article |
| id | doaj-art-04a6b8efbe914b18a571bff59ead89dd |
| institution | Kabale University |
| issn | 1663-9812 |
| language | English |
| publishDate | 2025-01-01 |
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| series | Frontiers in Pharmacology |
| spelling | doaj-art-04a6b8efbe914b18a571bff59ead89dd2025-01-30T09:26:22ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-01-011610.3389/fphar.2025.15046831504683αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia strokeYonghuan Yan0Linfang Wu1Lu Wang2Dandan Wang3Mengting Huang4Jinyong Peng5Jinyong Peng6Yingying Huang7Yingying Huang8Yingying Huang9School of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaCollege of Pharmacy, Dalian Medical University, Dalian, ChinaSchool of Pharmacy, Anhui University of Chinese Medicine, Hefei, ChinaAnhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, ChinaInstitute for the Evaluation of the Efficacy and Safety of Chinese Medicines, Anhui Academy of Chinese Medicine, Hefei, ChinaIntroductionαAsarone, an essential oil derived from Acorus gramineus Aiton, which has been successfully used to treat epilepsy in traditional chinese medicine, and has also been reported to confer neuroprotective effects on stroke. However, its mechanism of action remains poorly understood.MethodsThe effects of αAsarone on autophagy were examined by WB, RT-qPCR, immunofluorescence colocalization, transmission electron microscope, and autophagic flux activity was measured by infecting HT22 cells with mRFP-GFP-LC3 adenovirus. And then, cells were transfected with both mimic-miR-499-5p and inhibit-miR-499-5p to investigate the role of miR-499-5p in regulating the effects of αAsarone on stroke. To further clarify the protective effect of αAsarone in vivo, TTC staining, neurological function score, H&E staining, Nissl staining, Laser speckle contrast imaging, transmission electron microscopy, immunofluorescence colocalization, WB and RT-qPCR were performed in the MCAO mice.ResultsαAsarone was observed to inhibit the apoptosis of neuronal cells, and enhance autophagy. In addition, αAsarone promoted the expression of miR-499-5p. Targeting miR-499-5p can negatively regulate PDCD4 expression and the results from the dual-luciferase reporter assay demonstrate the direct targeting of PDCD4 by miR-499-5p. Promoting miR-499-5p can decrease the expression of PDCD4, increase ATG5, and enhance the protective effect of αAsarone on OGD/R injury while inhibiting miR-499-5p can weaken the effect of αAsarone. In vivo experiments further confirmed that αAsarone improved mice MCAO as evidenced by the amelioration of the neurological deficits and facilitated neuronal autophagy. Furthermore, we found that αAsarone reversed the effect of chloroquine, an autophagy inhibitor, and enhanced neuronal autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway.DiscussionOur data suggest that αAsarone alleviates neuronal injury of stroke by facilitating neuronal autophagy through the miR-499-5p/PDCD4/ATG5 signaling pathway.https://www.frontiersin.org/articles/10.3389/fphar.2025.1504683/fullautophagyneuronal injuryαAsaronestrokemiR-499-5p |
| spellingShingle | Yonghuan Yan Linfang Wu Lu Wang Dandan Wang Mengting Huang Jinyong Peng Jinyong Peng Yingying Huang Yingying Huang Yingying Huang αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke Frontiers in Pharmacology autophagy neuronal injury αAsarone stroke miR-499-5p |
| title | αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke |
| title_full | αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke |
| title_fullStr | αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke |
| title_full_unstemmed | αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke |
| title_short | αAsarone alleviates neuronal injury by facilitating autophagy via miR-499-5p/PDCD4/ATG5 signaling pathway in ischemia stroke |
| title_sort | αasarone alleviates neuronal injury by facilitating autophagy via mir 499 5p pdcd4 atg5 signaling pathway in ischemia stroke |
| topic | autophagy neuronal injury αAsarone stroke miR-499-5p |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2025.1504683/full |
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