N6-methyladenosine profiling reveals that Xuefu Zhuyu decoction upregulates METTL14 and BDNF in a rat model of traumatic brain injury

Dandan Feng; Pengfei Li; Wei Xiao; Zhuan Pei; Peishun Chen; Mingrui Hu; Zhaoyu Yang; Teng Li; Zian Xia; Hanjin Cui; Haigang Li; Qing Huang; Wei Zhang; Tao Tang; Yang Wang

doi:10.1016/j.jep.2023.116823

N⁶-methyladenosine profiling reveals that Xuefu Zhuyu decoction upregulates METTL14 and BDNF in a rat model of traumatic brain injury

J Ethnopharmacol. 2023 Dec 5:317:116823. doi: 10.1016/j.jep.2023.116823. Epub 2023 Jun 20.

Authors

Dandan Feng¹, Pengfei Li², Wei Xiao¹, Zhuan Pei¹, Peishun Chen¹, Mingrui Hu¹, Zhaoyu Yang¹, Teng Li¹, Zian Xia¹, Hanjin Cui¹, Haigang Li³, Qing Huang⁴, Wei Zhang⁵, Tao Tang¹, Yang Wang⁶

Affiliations

¹ Institute of Integrative Medicine, Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, Changsha, 410008, PR China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, 410008, PR China.
² Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
³ Hunan Key Laboratory of the Research and Development of Novel Pharmaceutical Preparations, Changsha Medical University, Changsha, 410219, PR China.
⁴ National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, 410008, PR China; Department of Neurology, Xiangya Hospital, Central South University, Changsha, 410008, PR China.
⁵ The Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, College of Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, 410208, PR China.
⁶ Institute of Integrative Medicine, Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, Changsha, 410008, PR China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, 410008, PR China. Electronic address: wangyang_xy87@csu.edu.cn.

PMID: 37348798
DOI: 10.1016/j.jep.2023.116823

Abstract

Ethnopharmacological relevance: The traditional Chinese herbal formula Xuefu Zhuyu decoction (XFZYD) is a classic formula in the category of invigorating blood circulation and resolving blood stasis. It has been proven to improve the neurological and ethological prognosis of traumatic brain injury. XFZYD promotes synaptic and axonal regeneration after traumatic brain injury, which is functionally modulated by the N⁶-methyladenosine (m⁶A) modification of RNA. However, the epigenetic effects of XFZYD on m⁶A modification remain unknown.

Aim of the study: To explore how XFZYD protects against traumatic brain injury induced by controlled cortical impact (CCI) injury by altering RNA m⁶A modification.

Materials and methods: The modified neurological severity scoring and Morris water maze were performed to evaluate the neuroprotective effects of XFZYD for 14 days and screen the dose. Then, dot blot, western blotting, and methylated RNA immunoprecipitation sequencing (MeRIP-Seq) were used to explore changes in RNA m⁶A modification in the perilesional cortex. The Metascape platform was used to analyze the Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome pathway of the differential m⁶A-tagged genes. Furthermore, MeRIP-qPCR was conducted to quantify differences in the hub differential m⁶A modification gene brain-derived neurotrophic factor (Bdnf).

Results: XFZYD significantly ameliorated the neurological deficits, spatial learning, and memory impairments in rats post-CCI on day 14. XFZYD enhanced the m⁶A level, and the expression of METTL14 and YTHDC2 in the perilesional cortex of CCI rats. In all three groups, the 3'-untranslated regions and coding sequence were primarily enriched for m⁶A peaks. XFZYD reversed the increased proportion of 3'-untranslated regions, and the decreased proportion of coding sequence and 5'-untranslated regions post-CCI. Moreover, XFZYD markedly downregulated 41 elevated m⁶A-tagged transcripts and upregulated 119 decreased m⁶A-tagged transcripts following CCI. Gene ontology and KEGG pathway analysis revealed that XFZYD-regulated m⁶A-tagged transcripts were predominantly enriched in synapse assembly, synaptic plasticity, learning or memory, and MAPK signaling pathway. Then, the hub-regulated m⁶A-tagged gene BDNF was identified. Both the m⁶A methylation level and the protein level of BDNF were ascended by XFZYD treatment.

Conclusion: XFZYD improves neurological deficits, spatial learning and memory impairments in rats post-TBI probably through increasing the expression of METTL14 and BDNF in the cortex. Our study highlights a novel post-transcriptional regulation mechanism mediated by herbal medicine for traumatic brain injury treatment.

Keywords: Brain-derived neurotrophic factor; Epitranscriptional regulation; METTL14; N(6)-methyladenosine; Traumatic brain injury; Xuefu Zhuyu decoction.

MeSH terms

Animals
Brain Injuries, Traumatic* / drug therapy
Brain Injuries, Traumatic* / genetics
Brain Injuries, Traumatic* / metabolism
Brain-Derived Neurotrophic Factor* / genetics
RNA / therapeutic use
Rats
Untranslated Regions

Substances

Xue-Fu-Zhu-Yu decoction
Brain-Derived Neurotrophic Factor
N-methyladenosine
RNA
Untranslated Regions