Single-cell transcriptomic dissection of the cellular and molecular events underlying the triclosan-induced liver fibrosis in mice

Bai，Yun Meng1; Yang，Fan2,3; Luo，Piao2; Xie，Lu Lin2; Chen，Jun Hui1; Guan，Yu Dong2; Zhou，Hong Chao2; Xu，Teng Fei2; Hao，Hui Wen2; Chen，Bing2,3; Zhao，Jia Hui2,3; Liang，Cai Ling2; Dai，Ling Yun1,2; Geng，Qing Shan1,2; Wang，Ji Gang1,2,4,5,6

2023-12-01

10.1186/s40779-023-00441-3

Military Medical Research   影响因子和分区

2095-7467

2054-9369

Background: Triclosan [5-chloro-2-(2,4-dichlorophenoxy) phenol, TCS], a common antimicrobial additive in many personal care and health care products, is frequently detected in human blood and urine. Therefore, it has been considered an emerging and potentially toxic pollutant in recent years. Long-term exposure to TCS has been suggested to exert endocrine disruption effects, and promote liver fibrogenesis and tumorigenesis. This study was aimed at clarifying the underlying cellular and molecular mechanisms of hepatotoxicity effect of TCS at the initiation stage. Methods: C57BL/6 mice were exposed to different dosages of TCS for 2 weeks and the organ toxicity was evaluated by various measurements including complete blood count, histological analysis and TCS quantification. Single cell RNA sequencing (scRNA-seq) was then carried out on TCS- or mock-treated mouse livers to delineate the TCS-induced hepatotoxicity. The acquired single-cell transcriptomic data were analyzed from different aspects including differential gene expression, transcription factor (TF) regulatory network, pseudotime trajectory, and cellular communication, to systematically dissect the molecular and cellular events after TCS exposure. To verify the TCS-induced liver fibrosis, the expression levels of key fibrogenic proteins were examined by Western blotting, immunofluorescence, Masson’s trichrome and Sirius red staining. In addition, normal hepatocyte cell MIHA and hepatic stellate cell LX-2 were used as in vitro cell models to experimentally validate the effects of TCS by immunological, proteomic and metabolomic technologies. Results: We established a relatively short term TCS exposure murine model and found the TCS mainly accumulated in the liver. The scRNA-seq performed on the livers of the TCS-treated and control group profiled the gene expressions of > 76,000 cells belonging to 13 major cell types. Among these types, hepatocytes and hepatic stellate cells (HSCs) were significantly increased in TCS-treated group. We found that TCS promoted fibrosis-associated proliferation of hepatocytes, in which Gata2 and Mef2c are the key driving TFs. Our data also suggested that TCS induced the proliferation and activation of HSCs, which was experimentally verified in both liver tissue and cell model. In addition, other changes including the dysfunction and capillarization of endothelial cells, an increase of fibrotic characteristics in B plasma cells, and M2 phenotype-skewing of macrophage cells, were also deduced from the scRNA-seq analysis, and these changes are likely to contribute to the progression of liver fibrosis. Lastly, the key differential ligand-receptor pairs involved in cellular communications were identified and we confirmed the role of GAS6_AXL interaction-mediated cellular communication in promoting liver fibrosis. Conclusions: TCS modulates the cellular activities and fates of several specific cell types (including hepatocytes, HSCs, endothelial cells, B cells, Kupffer cells and liver capsular macrophages) in the liver, and regulates the ligand-receptor interactions between these cells, thereby promoting the proliferation and activation of HSCs, leading to liver fibrosis. Overall, we provide the first comprehensive single-cell atlas of mouse livers in response to TCS and delineate the key cellular and molecular processes involved in TCS-induced hepatotoxicity and fibrosis.

Hepatic stellate cell Liver fibrogenesis scRNA-seq Triclosan

SCI

英语

第一

National Key Research and Development Program of China["2020YFA0908000","2022YFC2303600"] ; Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine[ZYYCXTD-C-202002] ; National Natural Science Foundation of China["82141001","82274182","82173914","82074098","81903588","82003814"] ; Science and Technology Foundation of Shenzhen[JCYJ20210324115800001] ; Fundamental Research Funds for the Central Public Welfare Research Institutes[ZXKT18003] ; Fundamental Research Funds for the Central public welfare research institutes[ZZ14-YQ-050] ; National Key R&D Program of China Key projects for international cooperation on science, technology and innovation[2020YFE0205100] ; Shenzhen Governmental Sustainable Development Fund[KCXFZ20201221173612034] ; Shenzhen key Laboratory of Kidney Diseases[ZDSYS201504301616234] ; Shenzhen Fund for Guangdong Provincial High-level Clinical Key Specialties[SZGSP001] ; Shenzhen Fund for Guangdong Provincial High-level Clinical Key Specialties[SZSM201612034]

General & Internal Medicine

Medicine, General & Internal

WOS:000936873700001

BMC

2-s2.0-85148548956

Scopus

1.Department of Nephrology,Shenzhen Key Laboratory of Kidney Diseases,and Shenzhen Clinical Research Centre for Geriatrics,Shenzhen People’s Hospital,the First Affiliated Hospital,Southern University of Science and Technology,Shenzhen,518020,China
2.Department of Urology,Shenzhen People’s Hospital,the First Affiliated Hospital,Southern University Science and Technology,the Second Clinical Medical College,Jinan University,Shenzhen,518020,China
3.Integrated Chinese and Western Medicine Postdoctoral Research Station,Jinan University,Guangzhou,510632,China
4.Artemisinin Research Center,and Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing,100700,China
5.Guangdong Provincial Key Laboratory of New Drug Screening,School of Pharmaceutical Sciences,Southern Medical University,Guangzhou,510515,China
6.Center for Reproductive Medicine,Dongguan Maternal and Child Health Care Hospital,Southern Medical University,Dongguan,Guangdong,523125,China

Bai，Yun Meng,Yang，Fan,Luo，Piao,et al. Single-cell transcriptomic dissection of the cellular and molecular events underlying the triclosan-induced liver fibrosis in mice[J]. Military Medical Research,2023,10(1).

Bai，Yun Meng.,Yang，Fan.,Luo，Piao.,Xie，Lu Lin.,Chen，Jun Hui.,...&Wang，Ji Gang.(2023).Single-cell transcriptomic dissection of the cellular and molecular events underlying the triclosan-induced liver fibrosis in mice.Military Medical Research,10(1).

Bai，Yun Meng,et al."Single-cell transcriptomic dissection of the cellular and molecular events underlying the triclosan-induced liver fibrosis in mice".Military Medical Research 10.1(2023).