Z-ligustilide alleviates atherosclerosis by reconstructing gut microbiota and sustaining gut barrier integrity through activation of cannabinoid receptor 2

Si-Jing Liu; Jiao-Jiao Fu; Zhen-Yue Liao; Yi-Xin Liu; Jing He; Li-Ying He; Jing Bai; Jing-Yan Yang; Shu-Qi Niu; Jin-Lin Guo

doi:10.1016/j.phymed.2024.156117

Z-ligustilide alleviates atherosclerosis by reconstructing gut microbiota and sustaining gut barrier integrity through activation of cannabinoid receptor 2

Phytomedicine. 2024 Oct 3:135:156117. doi: 10.1016/j.phymed.2024.156117. Online ahead of print.

Authors

Si-Jing Liu¹, Jiao-Jiao Fu¹, Zhen-Yue Liao¹, Yi-Xin Liu¹, Jing He¹, Li-Ying He², Jing Bai¹, Jing-Yan Yang³, Shu-Qi Niu⁴, Jin-Lin Guo⁵

Affiliations

¹ College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu, 611103, China; Chongqing Key Laboratory of Sichuan-Chongqing Co-construction for Diagnosis and Treatment of Infectious Diseases Integrated Traditional Chinese and Western Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611130, China.
² Key Laboratory of Characteristic Chinese Medicine Resources in Southwest China, College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611103, China.
³ College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu, 611103, China; Chongqing Key Laboratory of Sichuan-Chongqing Co-construction for Diagnosis and Treatment of Infectious Diseases Integrated Traditional Chinese and Western Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611130, China. Electronic address: wojaoyjy@cdutcm.edu.cn.
⁴ College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu, 611103, China; Chongqing Key Laboratory of Sichuan-Chongqing Co-construction for Diagnosis and Treatment of Infectious Diseases Integrated Traditional Chinese and Western Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611130, China. Electronic address: niushuqi@cdutcm.edu.cn.
⁵ College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Chengdu, 611103, China; Chongqing Key Laboratory of Sichuan-Chongqing Co-construction for Diagnosis and Treatment of Infectious Diseases Integrated Traditional Chinese and Western Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611130, China; Key Laboratory of Characteristic Chinese Medicine Resources in Southwest China, College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611103, China. Electronic address: guo596@cdutcm.edu.cn.

PMID: 39426255
DOI: 10.1016/j.phymed.2024.156117

Abstract

Background: Z-Ligustilide (ZL) is an essential phthalide found in Ligusticum chuanxiong Hort, a commonly used traditional Chinese medicine for treating atherosclerosis (AS) clinically. ZL has been shown to be effective in treating AS. However, the underlying mechanism of ZL against AS and its potential targets remain elusive.

Purpose: The purpose of this research was to assess the influence of ZL on AS and explore the role of the gut microbiome in mediating this effect.

Methods: A well-established AS mouse model, apolipoprotein E deficient (ApoE^-/-) mice was used to examine the effects of ZL on AS, inflammation, and the intestinal barrier. To analyze the changes in gut microbial community, we employed the 16S rRNA gene sequencing. Antibiotic cocktail and fecal microbiota transplantation (FMT) were employed to clarify the contribution of the gut microbiota to the anti-AS effects of ZL. The mechanism through which ZL provided protective effects on AS and the intestinal barrier was explored by untargeted metabolomics, as well as by validating the involvement of cannabinoid receptor 2 (CB2R) in mice and Caco-2 cells.

Results: Oral administration of ZL inhibited the development of atherosclerotic lesions, improved plaque stability, inhibited the increase in serum and atherosclerotic inflammation, and improved intestinal barrier function. Fecal bacteria from ZL-treated mice induced similar beneficial effects on AS and the intestinal barrier. We used 16S RNA gene sequencing to reveal a significant increase in Rikenella abundance in both ZL-treated mice and ZL-FMT mice, which was associated with the beneficial effects of ZL. Further function prediction analysis of the gut microbiota and CB2R antagonist intervention experiment in mice and Caco-2 cells showed that the activation of CB2R resulted in the enhancement of the intestinal barrier by ZL. Furthermore, the analysis of metabolomic profiling revealed the enrichment of capsaicin upon ZL treatment, which induced the activation of CB2R in human colon epithelial cells.

Conclusion: Our study is the first to demonstrate that oral treatment with ZL has the potential to alleviate AS by reducing inflammation levels and enhancing the intestinal barrier function. This mechanism relies on the gut microbiota in a CB2R-dependent manner, suggesting promising strategies and ideas for managing AS. This study provides insights into a novel mechanism for treating AS with ZL.

Keywords: Atherosclerosis; Cannabinoid receptor 2; Gut barrier; Gut microbiota; Z-ligustilide.