Estrogen cholestasis induces gut and liver injury in rats involving in activating PI3K/Akt and MAPK signaling pathways
Introduction
Intrahepatic cholestasis results from an impairment of bile secretion and leads to intrahepatic retention of toxic bile constituents, which is a common manifestation of several liver diseases. The etiologies and pathogenesis of intrahepatic cholestasis are complicated, including infection, heredity, environment, pregnancy, and diet [1]. In recent years, however, studies have indicated that intestinal damages are observed in patients with cholestatic liver diseases, such as obstructive jaundice, primary sclerosing cholangitis (PSC) and primary biliary cholangitis (PBC) [2]. It has been reported that approximately 70%–80% of PSC patients represent extraintestinal manifestations of inflammatory bowel disease [3]. Besides, intestinal damages are also observed in bile duct ligation (BDL)- and CCl4-induced cholestatic animal models [4]. The close association between liver and intestine highlights the importance of the gut-liver axis in the progression of cholestasis.
Estrogen is a female sex hormone, which plays a critical role in the menstrual cycle and reproductive system. Furthermore, estrogen is also a major component in many drugs, such as oral contraceptives and hormone replacement medications [5]. However, estrogen and its metabolites are causally associated with intrahepatic cholestasis, especially in women who are pregnant (intrahepatic cholestasis of pregnancy, ICP), receiving oral contraceptive administration and postmenopausal hormone replacement therapy [6]. It has been demonstrated that estrogen can repress bile acid secretion through the ERα/FXR/BSEP pathway, which results in excessive accumulation of toxic bile acid in hepatocytes and leads to estrogen cholestasis (EC) [7]. EC is mainly characterized by high estrogen level and hepatic toxic bile acid accumulation, which induce liver injury through activating multiple cell signaling pathways. A recent study has reported that some ICP patients displayed unexplained gastrointestinal discomfort and intestinal leakage [8]. Moreover, women with a history of ICP during pregnancy are more likely to die from intestinal diseases later in life [9]. We have recently provided preliminary evidence suggesting that intestinal damages present in estrogen-induced cholestatic rats [10]. However, details about pathological conditions of EC-induced gut and liver injury are largely unknown and need to be confirmed.
Considering human relevance, experimental cholestasis induced by 17α-ethynylestradiol (EE) administration has been widely used to investigate the molecular mechanisms involved in EC [6]. In this work, we aim to gain insight into EC-induced gut and liver injury and the cell signaling implicated. Overall, our data indicate that EC induces liver inflammation, both gut and liver oxidative stress and apoptosis, and the activation of PI3K/Akt and MAPK signaling pathways represents important mechanisms underlying EC-induced gut and liver injury.
Section snippets
Materials
EE was purchased from Sigma (St. Louis, MO, USA). Antibodies against NF-κB, p-NF-κB, Iκbα p-Iκbα, Bax, Caspsae3, Cleaved Caspsae3, Akt, p-Akt, ERK1/2, p-ERK1/2, JNK, p-JNK, p38, and p-p38 were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibody against Bcl-2 was obtained from R&D Systems (Minneapolis, MN, USA). RIPA lysis buffer and protease inhibitor cocktail were obtained from Beyotime Biotechnology (Jiangsu, China). All other reagents were of analytical grade.
Animal experiments
Male
The rat model of EC as well as EC-induced gut and liver injury
The body weight of rats treated with EE was dose-dependent decreased compared with the control group (Fig. 1A). Liver weight and liver/body weight were significantly increased after EE treatment (Fig. 1A). ALP, γ-GT, and TBA are biomarkers to indicate cholestasis, and all of them were dramatically increased after EE treatment (Fig. 1B). These results suggest that we have successfully established an estrogen cholestatic model which can be used for further investigation.
Histological abnormalities
Discussion
Intrahepatic cholestasis derives from the accumulation of hepatic bile acids and disruption of bile secretion caused by various factors, gradually progressed to liver injury, fibrosis, cirrhosis, and even liver cancer [1]. In addition to liver damage, recent studies have suggested the intestine is also implicated [3]. Although the gut-liver injury in partial types of cholestasis is confirmed, however, the pathological conditions of EC-induced gut and liver injury remain largely unknown. Our
Funding
This work was supported by the National Natural Science Foundation of China (NO.81803798, NO.82073939, and NO.81573788) and the Tongji Hospital Fund (2019A08).
Declaration of conflicting interest
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
References (39)
- et al.
Primary sclerosing cholangitis
Lancet
(2018) - et al.
Bacterial translocation and changes in the intestinal microbiome in mouse models of liver disease
J. Hepatol.
(2012) - et al.
The ileum-liver Farnesoid X receptor signaling axis mediates the compensatory mechanism of 17α-ethynylestradiol-induced cholestasis via increasing hepatic biosynthesis of chenodeoxycholic acids in rats
Eur. J. Pharm. Sci.
(2018) - et al.
Taxifolin ameliorates iron overload-induced hepatocellular injury: modulating PI3K/AKT and p38 MAPK signaling, inflammatory response, and hepatocellular regeneration
Chem. Biol. Interact.
(2020) - et al.
Emerging roles of bile acids in mucosal immunity and inflammation
Mucosal Immunol.
(2019) - et al.
Intestinal estrogen receptor beta suppresses colon inflammation and tumorigenesis in both sexes
Cancer Lett.
(2020) - et al.
Estrogen receptor beta activation ameliorates DSS-induced chronic colitis by inhibiting inflammation and promoting Treg differentiation
Int. Immunopharmacol.
(2019) - et al.
Akt promotes cell survival by phosphorylating and inhibiting a forkhead transcription factor
Cell
(1999) - et al.
Insulin/IGF-1 paradox of aging: regulation via AKT/IKK/NF-kappaB signaling
Cell. Signal.
(2010) - et al.
Inhibition of p38 and ERK1/2 pathways by Sparstolonin B suppresses inflammation-induced melanoma metastasis
Biomed. Pharmacother.
(2018)
Quercetin protects human hepatocytes from ethanol-derived oxidative stress by inducing heme oxygenase-1 via the MAPK/Nrf2 pathways
J. Hepatol.
New therapeutic concepts in bile acid transport and signaling for management of cholestasis
Hepatology
Novel aspects in the management of cholestatic liver diseases
Dig. Dis.
Hormonal contraceptives, female sexual dysfunction, and managing strategies: a review
J. Clin. Med.
Hepatic gene transfer of human aquaporin-1 improves bile salt secretory failure in rats with estrogen-induced cholestasis
Hepatology
Transcriptional dynamics of bile salt export pump during pregnancy: mechanisms and implications in intrahepatic cholestasis of pregnancy
Hepatology
Is a leaky gut involved in the pathogenesis of intrahepatic cholestasis of pregnancy?
Hepatology
Intrahepatic cholestasis of pregnancy and associated causes of death: a cohort study with follow-up of 27-46 years
BMC Womens Health
Intestinal injury in the rat model of 17α-ethynylestradiol-induced intrahepatic cholestasis
J. Dig. Dis.
Cited by (9)
Vitamin D Receptor Activation Reduces Hepatic Inflammation via Enhancing Macrophage Autophagy in Cholestatic Mice
2024, American Journal of Pathology“Yajieshaba” prevents acute alcoholic liver injury and repairs the intestinal mucosal barrier
2024, Journal of EthnopharmacologyCalculus Bovis Sativus alleviates estrogen cholestasis-induced gut and liver injury in rats by regulating inflammation, oxidative stress, apoptosis, and bile acid profiles
2023, Journal of EthnopharmacologyCitation Excerpt :Studies have reported that the serum levels of M30, a marker of apoptotic cell death, which indicates the caspase-cleaved cytokeratin-18 fragments, are dramatically higher in ICP and PBC patients than in healthy people (Sekiguchi et al., 2015; Ersoy et al., 2016). In animal models, such as bile duct ligation- and estrogen-induced cholestasis, cell apoptosis could not only be observed in the liver but also in the intestine (Runping Liu et al., 2018; Xiang et al., 2021). Cell apoptosis, known as programmed cell death, is modulated by kinds of regulatory proteins.
Vitamin D Receptor Activation Targets ROS-Mediated Crosstalk Between Autophagy and Apoptosis in Hepatocytes in Cholestasic Mice
2023, Cellular and Molecular Gastroenterology and HepatologyValidation of MAPK signalling pathway as a key role of paeoniflorin in the treatment of intrahepatic cholestasis of pregnancy based on network pharmacology and metabolomics
2022, European Journal of PharmacologyCitation Excerpt :MAPK signalling pathway is mainly composed of the extracellular signal-regulated kinase (ERK), c-Jun-N-terminal kinase (JNK), and p38 (Sun et al., 2015). MAPK signalling pathway has been shown to play an important role during cholestatic liver injury (Yuan et al., 2020; Xiang et al., 2021). Our results showed that PF was found to significantly reduce the relative protein expression of p-ERK, p-JNK, and p-P38 in ICP, indicating that PF can inhibit the activation of the MAPK pathway.
A new strategy for the rapid identification and validation of direct toxicity targets of psoralen-induced hepatotoxicity
2022, Toxicology LettersCitation Excerpt :Finally, by combining information from the literature, we confirmed that the core targets ABL1, mTOR, and Nrf2 were the main nodes (Supplementary Table 6). The PI3K-AKT-Nrf2-mTOR signaling pathway is involved in the occurrence of chronic liver injury, liver cirrhosis, liver failure, and other liver diseases (Xiang et al., 2021). The ABL1 inhibitor imatinib mesylate can induce myocardial cell injury in vitro, and its mechanism is related to mitochondrial membrane disturbance, high ROS levels, the downregulation of Nrf2 expression, attenuated GSH activity, and cardiac damage through the regulation of the Nrf2 pathway (Song et al., 2022).
- 1
These authors contributed equally to this study.