Danggui-Shaoyao San alleviates cognitive impairment via enhancing HIF-1α/EPO axis in vascular dementia rats

Yuan Ningning; Xu Ying; Li Xiang; Su Yue; Wang Zhongda; Jiang Ruoyu; Shi Hanwen; Tao Weiwei; Zhang Yafeng; Ma Junjie; Cheng Xiaolan

doi:10.1016/j.jep.2024.118306

Danggui-Shaoyao San alleviates cognitive impairment via enhancing HIF-1α/EPO axis in vascular dementia rats

J Ethnopharmacol. 2024 May 7:331:118306. doi: 10.1016/j.jep.2024.118306. Online ahead of print.

Authors

Yuan Ningning¹, Xu Ying¹, Li Xiang¹, Su Yue¹, Wang Zhongda¹, Jiang Ruoyu¹, Shi Hanwen¹, Tao Weiwei¹, Zhang Yafeng², Ma Junjie³, Cheng Xiaolan⁴

Affiliations

¹ School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
² Jiangsu CM Clinical Innovation Center of Degenerative Bone & Joint Disease, Wuxi TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Wuxi, 214071, China. Electronic address: orthozhang@sina.com.
³ School of Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China. Electronic address: 290407@njucm.edu.cn.
⁴ School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China. Electronic address: chengxiaolan37@njucm.edu.cn.

PMID: 38723920
DOI: 10.1016/j.jep.2024.118306

Abstract

Ethnopharmacological relevance: Invigorating blood circulation to remove blood stasis is a primary strategy in TCM for treating vascular dementia (VaD). Danggui-Shaoyao San (DSS), as a traditional prescription for neuroprotective activity, has been proved to be effective in VaD treatment. However, its precise molecular mechanisms remain incompletely understood.

Aim of the study: The specific mechanism underlying the therapeutic effects of DSS on VaD was explored by employing network pharmacology as well as in vivo and in viro experiment validation.

Materials and methods: We downloaded components of DSS from the BATMAN-TCM database for target prediction. The intersection between the components of DSS and targets, PPI network, as well as GO and KEGG enrichment analysis were then performed. Subsequently, the potential mechanism of DSS predicted by network pharmacology was assessed and validated through VaD rat model induced by 2VO operation and CoCl₂-treated PC12 cells. Briefly, the DSS extract were first quantified by HPLC. Secondly, the effect of DSS on VaD was studied using MWM test, HE staining and TUNEL assay. Finally, the molecular mechanism of DSS against VaD was validated by Western blot and RT-QPCR experiments.

Results: Through network analysis, 137 active ingredients were obtained from DSS, and 67 potential targets associated with DSS and VaD were identified. GO and KEGG analysis indicated that the action of DSS on VaD primarily involves hypoxic terms and HIF-1 pathway. In vivo validation, cognitive impairment and neuron mortality were markedly ameliorated by DSS. Additionally, DSS significantly reduced the expression of proteins related to synaptic plasticity and neuron apoptosis including PSD-95, SYP, Caspase-3 and BCL-2. Mechanistically, we confirmed DSS positively modulated the expression of HIF-1α and its downstream proteins including EPO, p-EPOR, STAT5, EPOR, and AKT1 in the hippocampus of VaD rats as well as CoCl₂-induced PC12 cells. HIF-1 inhibitor YC-1 significantly diminished the protection of DSS on CoCl₂-induced PC12 cell damage, with decreased HIF-1α, EPO, EPOR expression.

Conclusion: Our results initially demonstrated DSS could exert neuroprotective effects in VaD. The pharmacological mechanism of DSS may be related to its positive regulation on HIF-1α/EPO pathway.

Keywords: Danggui-Shaoyao San; EPO; HIF-1α; Network pharmacology; Vascular dementia.