Single-cell transcriptomic insights into endosulfan-induced liver injury: Key pathways and inflammatory responses

Pan Huang; Yunmeng Bai; Chaohua Zhou; Xiaoru Zhong; Ashok Iyaswamy; Peng Chen; XuWei; Wei Zhang; Chuanbin Yang; Jigang Wang

doi:10.1016/j.livres.2025.05.002

Liver Research ›› 2025, Vol. 9 ›› Issue (2) :144 -156. DOI: 10.1016/j.livres.2025.05.002

Research article

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Single-cell transcriptomic insights into endosulfan-induced liver injury: Key pathways and inflammatory responses

Pan Huang ^a^,¹
, Yunmeng Bai ^b^,¹
, Chaohua Zhou ^b^,¹
, Xiaoru Zhong ^b^,¹
, Ashok Iyaswamy ^c^,^d
, Peng Chen ^e
, XuWei ^f
, Wei Zhang ^g^,^h^,^*
, Chuanbin Yang ^b^,^**
, Jigang Wang ^a^,^b^,^g^,ⁱ^,^***

Author information +

^a Department of Epidemiology, School of Public Health, Guangxi Medical University, Nanning, Guangxi, China

^b Department of Critical Care Medicine, Guangdong Provincial Clinical Research Center for Geriatrics, Shenzhen Clinical Research Center for Geriatrics, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University);The First Afﬁliated Hospital, Southern University of Science and Technology), Shenzhen, Guangdong, China

^c Mr. and Mrs. Ko Chi-Ming Centre for Parkinson’s Disease Research, School of Chinese Medicine, Hong Kong Baptist University, Hong Kong SAR, China

^d Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore, India

^e Experimental Research Center, China Academy of Traditional Chinese Medicine, Beijing, China

^f Wangjing Hospital, China Academy of Chinese Medical Sciences, Beijing, China

^g Center for Drug Research and Development, Guangdong Provincial Key Laboratory for Research and Evaluation of Pharmaceutical Preparations, Guangdong Pharmaceutical University, Guangzhou, Guangdong, China

^h Department of Critical Care, The First Afﬁliated Hospital of Guangdong Pharmaceutical University, Guangzhou, China

ⁱ State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Artemisinin Research Center, and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China

^* Center for Drug Research and Development, Guangdong Provincial Key Laboratory for Research and Evaluation of Pharmaceutical Preparations, Guangdong Pharmaceutical University, Guangzhou, Guangdong, China. E-mail addresses: zhangweia@gdpu.edu.cn (Wei Zhang)

^** Department of Critical Care Medicine, Guangdong Provincial Clinical Research Center for Geriatrics, Shenzhen Clinical Research Center for Ge-riatrics, Shenzhen People’s Hospital; The First Afﬁliated Hospital, Southern University of Science and Technology, Shenzhen, Guangdong, China. E-mail addresses: h1094103@connect.hku.hk (Chuanbin Yang)

^*** State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Artemisinin Research Center, and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China. E-mail addresses: jgwang@icmm.ac.cn (Jigang Wang)

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Abstract

Background and aims: Environmental pollutants, particularly organochlorine insecticides like endosulfan (ENDO), are increasingly linked to liver toxicity and related diseases. Despite its widespread historical use, the mechanisms underlying ENDO-induced liver damage remain poorly understood. This study aims to elucidate the cellular and molecular mechanisms of ENDO-induced hepatotoxicity.

Methods: C57BL/6 mice were exposed to ENDO for two weeks. Single-cell RNA sequencing (scRNA-seq) was subsequently performed on mouse livers to explore ENDO-induced hepatotoxicity at the single-cell level. Differentially expressed genes (DEGs) across cell types and treatments were identified and then subjected to pathway enrichment to uncover key biological processes affected by ENDO. Transcription factor (TF) regulatory network, pseudotime trajectory, and cellular communication analysis were used to explore the molecular and cellular changes after ENDO exposure.

Results: ENDO not only caused direct hepatocyte injury but also activated hepatic stellate cells and lymphocytes, triggering inflammatory responses with upregulation of multiple key chemokines and cytotoxic genes. Additionally, ENDO exposure led to the recruitment and activation of myeloid cells, contributing to the inflammatory milieu. An increase in intercellular communication and changes to the hepatic microenvironment, especially the interaction between activated hepatic stellate cells and CD8⁺ T cells were observed, further implicating these processes in ENDO-induced liver damage.

Conclusions: This study provides new insights into the cellular and molecular mechanisms underlying liver injury induced by organochlorine insecticides like ENDO. Key genes and pathways involved in ENDO-associated liver toxicity have been identified at a single-cell resolution. These findings suggest that altered cellular communications and inflammatory responses may play pivotal roles in the pathogenesis of ENDO-induced liver injury.

Keywords

Endosulfan (ENDO) / Liver injury / hepatotoxicity / Single-cell RNA sequencing (scRNA-seq) / Inflammatory responses / Intercellular communication / Hepatic microenvironment

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Pan Huang, Yunmeng Bai, Chaohua Zhou, Xiaoru Zhong, Ashok Iyaswamy, Peng Chen, XuWei, Wei Zhang, Chuanbin Yang, Jigang Wang. Single-cell transcriptomic insights into endosulfan-induced liver injury: Key pathways and inflammatory responses. Liver Research, 2025, 9(2): 144-156 DOI:10.1016/j.livres.2025.05.002

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Authors' contributions

Pan Huang: Investigation, Writing eoriginal draft, review & editing. Yunmeng Bai: Methodology, Investigation, Data analysis, Methodology, Writing e review & editing. Chaohua Zhou: Con-ceptualization, Writing e review & editing, Resources. Xiaoru Zhong: Conceptualization, Writing e review & editing, Resources. Ashok Iyaswamy: Methodology, Writing e review & editing. Peng Chen: Methodology, Resources. Xu Wei: Methodology, Resources. Wei Zhang: Supervision, Conceptualization, Writing e review & editing, Finding. Chuanbin Yang: Supervision, Conceptualization, Writing e review & editing, Funding. Jigang Wang: Software, Su-pervision, Conceptualization, Writing e review & editing, Funding.

Data availability

Data are contained within the article or supplementary mate-rial. The data in the current study are available from the corre-sponding author upon request. The sequencing data reported in this study have been deposited in the OMIX, China National Center for Bioinformation/Beijing Institute of Genomics, Chinese Academy of Sciences (https://ngdc.cncb.ac.cn/omix/release/OMIX004105).

Declaration of competing interest

The authors declare that they have no conflict of interest.

Acknowledgments

The work was supported by grants from the National Key Research and Development Program of China (No. 2022YFC2303603 and 2020YFA0908004), the Shenzhen Science and Technology Program (Nos. JCYJ20220818102613029, JCYJ20240813175901003, and JCYJ20240813103823031), China Postdoctoral Science Foundation (No. 2024M752143), the Shenz-hen Medical Research Funds (No. B2302051), the Scientiﬁc and technological innovation project of China Academy of Chinese Medical Sciences (CI2023D003 and CI2023E005TS05), the CACMS Innovation Fund (CI2023E002 and ZG2024001-05), the Funda-mental Research Funds for the Central public welfare research in-stitutes (ZZ13-ZD-07, ZZ14-YQ-050, ZZ14-FL-010, ZZ14-ND-010, ZZ15-ND-10, ZZ16-ND-10-23, ZZ17-ND-10 and ZZ18-ND-10).

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.livres.2025.05.002.

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