Silver nanoparticles induced hepatoxicity via the apoptotic/antiapoptotic pathway with activation of TGFβ-1 and α-SMA triggered liver fibrosis in Sprague Dawley rats

Environmental Science and Pollution Research https://doi.org/10.1007/s11356-022-21388-3 RESEARCH ARTICLE Silver nanoparticles induced hepatoxicity via the apoptotic/ antiapoptotic pathway with activation of TGFβ‑1 and α‑SMA triggered liver fibrosis in Sprague Dawley rats Doaa H. Assar1 · Abd‑Allah A. Mokhbatly1 · Emad W. Ghazy1 · Zizy I. Elbialy2 Ayman A. Hassan3 · Ahmed Nabil4,5 · Samah Abou Asa6 · Ahmed A. Gaber1 · Received: 8 March 2022 / Accepted: 6 June 2022 © The Author(s) 2022 Abstract Despite the extraordinary use of silver nanoparticles (AgNPs) in medicinal purposes and the food industry, there is rising worry about potential hazards to human health and the environment. The existing study aims to assess the hepatotoxic effects of different dosages of AgNPs by evaluating hematobiochemical parameters, oxidative stress, liver morphological alterations, immunohistochemical staining, and gene expression to clarify the mechanism of AgNPs’ hepatic toxic potential. Forty male Sprague Dawley rats were randomly assigned into control and three AgNPs intraperitoneally treated groups 0.25, 0.5, and 1 mg/kg b.w. daily for 15 and 30 days. AgNP exposure reduced body weight, caused haematological abnormalities, and enhanced hepatic oxidative and nitrosative stress with depletion of the hepatic GSH level. Serum hepatic injury biomarkers with pathological hepatic lesions where cholangiopathy emerges as the main hepatic alteration in a dosage- and durationdependent manner were also elevated. Furthermore, immunohistochemical labelling of apoptotic markers demonstrated that Bcl-2 was significantly downregulated while caspase-3 was significantly upregulated. In conclusion, the hepatotoxic impact of AgNPs may be regulated by two mechanisms, implying the apoptotic/antiapoptotic pathway via raising BAX and inhibiting Bcl-2 expression levels in a dose-dependent manner. The TGF-β1 and α-SMA pathway which triggered fibrosis with incorporation of iNOS which consequently activates the inflammatory process were also elevated. To our knowledge, there has been no prior report on the experimental administration of AgNPs in three different dosages for short and long durations in rats with the assessment of Bcl-2, BAX, iNOS, TGF-β1, and α-SMA gene expressions. Keywords Silver nanoparticles · Hepatotoxicity · Fibrosis · Apoptotic pathway · TGFβ · α-SMA Responsible Editor: Lotfi Aleya * Zizy I. Elbialy Doaa H. Assar Abd‑Allah A. Mokhbatly Emad W. Ghazy Ahmed A. Gaber Ayman A. Hassan Ahmed Nabil 1 Clinical Pathology Department, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh 33516, Egypt 2 Department of Fish Processing and Biotechnology, Faculty of Aquatic and Fisheries Sciences, Kafrelsheikh University, Kafrelsheikh 33516, Egypt 3 High Technological Institute of Applied Health Sciences, Egypt Liver Research Institute and Hospital (ELRIAH), Sherbin, El Mansora, Egypt 4 Beni-Suef University, Beni‑Suef, Egypt 5 Egypt Liver Research Institute and Hospital (ELRIAH), Sherbin, El Mansora, Egypt 6 Pathology Department, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh 33516, Egypt Samah Abou Asa 13 Vol.:(0123456789) Environmental Science and Pollution Research Introduction Nanotechnology is widely used in various applications of nutrition, therapy and medication with incorporation into consumer products, for example, food packaging material, food supplements, textiles, and spray products, because of their antimicrobial properties as well as potent anti-inflammatory, antiviral and/or anticancer activities (Wei et al. 2015; Lee and Jun 2019; Paladini and Pollini 2019), making it likely that the biomedical applications of these nanoparticles (NPs) will expand in the future. Hence, increased consumer exposure to silver nanoparticle (AgNP)–containing products has raised the potential need for their toxicological assessment (Rosario et al. 2020). Several in vitro studies have shown that AgNPs can induce cytotoxicity, DNA damage, oxidative stress (OS) and inflammatory responses in human cells (Ahamed et al. 2008; Ahamed et al. 2010; Rosario et al. 2016; Carrola et al. 2016; Bastos et al. 2016; Akter et al. 2017; Rosario et al. 2018), but it still has not been clearly identified yet, and few studies have studied on the toxic effects of sublethal doses exposed to AgNPs for a short period (Lee et al. 2012; Lamberti et al. 2014). In earlier studies, Takenaka et al. (2001) and Arora et al. (2009) reported that the liver appears to be a major accumulation site of circulatory AgNPs, as a significant quantity of AgNPs is detected in the liver of rats following a 90-day oral administration (Kim et al. 2010). A recent clinical report also described the absorption of AgNPs into the circulation following the use of AgNP-coated dressings for burns (Vlachou et al. 2007; Ferdous and Nemmar 2020). Van der Zande et al. (2012) recorded that the liver is from the major organs of AgNPs (De Jong et al. 2013). The excessive accumulation of AgNPs in the liver led to severe pathological changes (Lee et al. 2013a, b). Also, AgNPs have an important role in reactive oxygen species (ROS) induction in many body cells (Choi et al. 2009). ROS are continually produced and removed in biological systems by endogenous or exogenous antioxidants (Mohammadi et al. 2013; Piao et al. 2011), but excessive generation of ROS can lead to apoptosis and cause oxidative DNA damage (Xu et al. 2012). The purpose of the existing study was to assess the impact of different doses of AgNPs on body performance, haemato-biochemical parameters, oxidative stress/ antioxidant status, hepatic morphological alterations, apoptotic/ antiapoptotic pathway and the underlying molecular mechanisms after 15 and 30 days of intraperitoneal injection of AgNPs. Materials and methods Ethical statement The experiment was approved by the Institutional Animal Care and Animal Ethics Committee, Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt. All precautions 13 were followed to diminish animal suffering during the experiment (KFS2020-3). Silver nanoparticle synthesis Silver nanoparticles were synthesized as follows: 25 mL of 6.8 mM trisodium citrate in an aqueous solution, containing 7 μM of tannic acid, was heated to 60 °C in an oil bath. After heating, the solution was added directly with strong stirring to 100 mL of 0.74 mM AgNO3 which was also pre-heated to 60 °C. This mixture was kept at 60 °C for a few minutes until the colour of the solution turned to yellow. The mixture was then kept at 97 °C for a further 45 min followed by cooling down to room temperature and finally stored in the dark at 4 °C (Bastús et al. 2014). Characterization of silver nanoparticles Characterization of AgNPs is important in order to evaluate the functional aspects of the synthesized particles. We characterize our prepared AgNPs by two different analytical techniques: dynamic light scattering (DLS) and transmission (...truncated)