Chemo-preventive effect of crocin against experimentally-induced hepatocarcinogenesis via regulation of apoptotic and Nrf2 signaling pathways
Introduction
Hepatocellular carcinoma (HCC) is the most common liver malignancy. It represents a global health concern due to various reasons, such as its high prevalence, its aggressive characteristics, and the generally poor prognosis (Li et al., 2019; Yang et al., 2019). The high angio-invasive capacity of the tumor, ease of metastasis, and multidrug resistance limit the efficacy of chemo and radio-therapeutic agents. Moreover, these treatment options show significant side effects, which significantly affect the patient’s quality of life (Lohitesh et al., 2018). Therefore, the current mainstay treatment for HCC is surgical resection. Nevertheless, the high cost, the shortage of matching donors, potential recurrence, and sub-optimal outcomes preclude this solution for many HCC patients (Chen et al., 2018). Thus, it has become crucial to develop alternative treatment strategies to control HCC development and progression, so reducing mortality and improving survival rates.
c-Jun N-terminal kinase (c-JNK) is a member of mitogen-activated protein kinase significantly implicated in hepatocarcinogenesis. Previous studies referred to the involvement of c-JNK signaling in the regulation of cancer proliferation via bypassing cellular apoptosis (Recio-Boiles et al., 2016). Indeed, antagonizing p53-induced apoptosis has been reported to be a major mechanism involved in liver tumor development (Eferl et al., 2003). Moreover, c-JNK inhibition in HCC cells increased caspase recruitment, arrested hepatocyte proliferation, and enhanced TRAIL-mediated apoptosis (Mucha et al., 2009).
Progression of HCC is associated with redox imbalance characterized by lipid peroxidation, defective cellular protective mechanisms, and interference of free radicals with signal transduction pathways. In this context, agents that can control oxidative stress by up-regulating endogenous protective mechanisms have been reported to be effective in hepatocarcinogenesis control (Takaki and Yamamoto, 2015).
Nuclear factor erythroid 2–related factor 2 (Nrf2) is a transcription factor that controls various cellular protective mechanisms. Oxidative stimuli induce Nrf2 signaling with the subsequent triggering of the antioxidant response element (ARE) transcription. This is followed by the transcriptional induction of multiple antioxidant proteins. Nrf2 dysregulation is concomitant with the early stages of tumorigenesis. Indeed, the Nrf2 gene and protein expression were down-regulated in hepatocytes of experimental models of HCC with a subsequent decrease in antioxidant defence mechanisms (Al-Gayyar et al., 2019).
Thioacetamide (TAA) is a powerful hepatotoxin and repeated exposure to TAA induces liver cirrhosis and carcinomas in various rodent models (Dwivedi and Jena, 2020). Crocin is a water-soluble dicarboxylic acid monoglyceride (carotenoid) derived from saffron, the dried dark-red stigma of Crocus sativus L. (Wang et al., 2018). Studies have reported multiple beneficial activities of crocin which were attributed to its antioxidant, anti‐inflammatory (Jnaneshwari et al., 2013) and immunity enhancement (Bolhassani et al., 2014) effects. Interestingly, various studies have reported crocin treatment to induce apoptosis in several types of cancer including skin (Wang et al., 2018), colorectal (Aung et al., 2007), breast (Arzi et al., 2018), cervical (Mollaei et al., 2017) cancers and gastric adenocarcinoma (Hoshyar et al., 2013). Interestingly, in HCC, crocin demonstrated anti-proliferative and pro-apoptotic effects both in vivo and in vitro (Amin et al., 2016).
Accordingly, the present study aimed to evaluate the hepatoprotective effect of crocin in a rat model of TAA-induced hepatocarcinogenesis with an emphasis on evaluation of the modulatory role of crocin on apoptotic signaling in HCC. The apoptotic effect of crocin through the alternation of TRAIL, p53, caspase 8, BAX, Bcl-2 and c-JNK as an upstream regulator as well as the effect of crocin treatment on Nrf2 signaling were investigated.
Section snippets
Animals
Thirty-six adult male Sprague Dawley rats weighing 180−200 g were provided from "Urology and Nephrology Center", Mansoura, Egypt. Rats were adapted under the standard nutritional and environmental conditions following the ethical principles and guidelines for the care and use and handling of experimental animals adopted by the Research Ethics Committee, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt.
Drugs and chemicals
Thioacetamide (TAA) and crocin were purchased from Sigma Aldrich, (St. Louis,
Results
By the end of weeks 8 and 12 of the experimental procedures, HCC rats revealed a persistent significant elevation in serum α fetoprotein levels compared to normal control, (suppl Fig. 1).
Discussion
Hepatocellular carcinoma (HCC) is one of the top causes of cancer-related deaths worldwide, particularly in developing countries (Abd-Elsalam et al., 2018; Yang et al., 2019). Over the past decade, progress has been shown in trying to understand HCC’s etiology, risk factors, and underlying molecular mechanisms. Indeed, different therapeutic approaches have been suggested, yet the rate of HCC occurrence continues to escalate with most patients presenting at later stages (Baecker et al., 2018;
CRediT authorship contribution statement
Nehal M. Elsherbiny: Conceptualization, Methodology, Software, Formal analysis, Investigation, Resources, Data curation, Writing - original draft, Writing - review & editing, Visualization, Project administration. Nada H. Eisa: Conceptualization, Methodology, Software, Investigation, Resources, Data curation, Writing - original draft, Writing - review & editing, Visualization, Project administration. Mohamed El-Sherbiny: Methodology, Software, Formal analysis, Investigation, Resources, Data
Declaration of Competing Interest
The authors report no declarations of interest.
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