Aspergillus awamori attenuates ochratoxin A-induced renal and cardiac injuries in rabbits by activating the Nrf2/HO-1 signaling pathway and downregulating IL1β, TNFα, and iNOS gene expressions

Environmental Science and Pollution Research https://doi.org/10.1007/s11356-022-20599-y RESEARCH ARTICLE Aspergillus awamori attenuates ochratoxin A‑induced renal and cardiac injuries in rabbits by activating the Nrf2/HO‑1 signaling pathway and downregulating IL1β, TNFα, and iNOS gene expressions Doaa H. Assar1 · Samah Abou Asa2 · Moshira A. El‑Abasy3 · Zizy I. Elbialy4 · Mustafa Shukry5 · Amera Abd El Latif6 · Mona N. BinMowyna7 · Norah A. Althobaiti8 · Mohammed A. El‑Magd9 Received: 22 March 2022 / Accepted: 29 April 2022 © The Author(s) 2022 Abstract Ochratoxin A (OTA) is one of the most dangerous and that pollute agricultural products, inducing a variety of toxic effects in humans and animals. The current study explored the protective effect of different concentrations of Aspergillus awamori (A. awamori) against OTA (0.3 mg/kg diet) induced renal and cardiac damage by exploring its mechanism of action in 60 New Zealand white male rabbits. Dietary supplementation of A. awamori at the selected doses of 50, 100, and 150 mg/kg diet, respectively, for 2 months significantly improved the rabbit’s growth performance; modulated the suppressed immune response and restored the altered hematological parameters; reduced the elevated levels of renal injury biomarkers such as urea, creatinine, and alkaline phosphatase; and increased serum total proteins concentrations. Moreover, it also declined enzymatic activities of cardiac injury biomarkers, including AST, LDH, and CK-MB. A. awamori alleviated OTA-induced degenerative and necrotic changes in the kidney and heart of rabbits. Interestingly, A. awamori upregulated Nrf2/OH-1 signaling pathway. Therefore enhanced TAC, CAT, and SOD enzyme activities and reduced OTA-induced oxidative and nitrosative stress by declining iNOS gene expression and consequently lowered MDA and NO levels. In addition to attenuating renal and cardiac inflammation via reducing IL-1β, TNF-α gene expressions in a dose-dependent response. In conclusion,this is the first report to pinpoint that dietary incorporation of A. awamori counteracted OTA-induced renal and cardiac damage by potentiating the rabbit’s antioxidant defense system through its potent antioxidant, free radical scavenging, and antiinflammatory properties in a dose-dependent response. Based on our observations, A. awamori could be utilized as a natural protective agent against ochratoxicosis in rabbits. Keywords Ochratoxin A · Aspergillus awamori · Oxidative stress · Histopathology · Gene expression Abbreviations A. awamori Aspergillus awamori AST Aspartate aminotransferase ALP Alkaline phosphatase CAT Catalase CK-MB Creatine kinase-MB TNF-α Tumor necrosis factor-alpha IL-1β Interleukin 1 beta HO-1 Haemoxygenase-1 iNOS Nitric oxide synthases LDH Lactate dehydrogenase Responsible Editor: Lotfi Aleya * Zizy I. Elbialy Extended author information available on the last page of the article Nrf2 Nuclear factor erythroid 2-related factor 2 MDA Malondialdehyde NO Nitric oxide OTA Ochratoxin A SOD Superoxide dismutase TAC Total antioxidant capacity Introduction Ochratoxins are secondary toxic metabolites produced by various fungi of the genus Aspergillus and Penicillium (Ostry et al. 2013) that cause several harmful influences on multiple animals (Pfohl-Leszkowicz and Manderville 2007; Battacone et al. 2010). Rabbits are one of the applied alternative sources to face the scarcity of meat resources in developing countries (Dalle Zotte and Szendrő, 2011). 13 Vol.:(0123456789) Environmental Science and Pollution Research Rabbit meat consumption is a rising economic industry routinely consumed in Egypt (Baviera-Puig et al., 2017). Rabbits are comparatively more vulnerable to Ochratoxin A (OTA) than mice, rats, and guinea pigs (Ponnuchamy, 2000). OTA has gained a special consideration among the recorded mycotoxins due to its nephrotoxic, teratogenic, embryotoxic, immunosuppressive, genotoxic, neurotoxic, and carcinogenic properties (O'Brien et al.2001). OTA has good thermal stability, making its eradication from the food chain impossible (Malir et al. 2016). Based on OTA nature, humans and domestic animals that are chronically exposed to low doses of OTA (50–250 μg OTA/kg b.m.) are potentially at risk for renal diseases or may even exhibit carcinogenic possibilities (Abdel-Wahhab et al. 2005; Ringot et al. 2006; Brown et al. 2007; Zain, 2011; Gruber-Dorninger et al. 2019). The most relevant impacts of OTA are the nephrotoxicity and nephron-carcinogenicity in rodents (Benford et al., 2001). The high sensitivity of kidneys to OTA can be attributed to the kidney's critical importance as an exclusive excretory organ for OTA elimination (Marquardt and Frohlich, 1992). Moreover, the exact mechanisms of its toxicity are not fully understood. Among these mechanisms, oxidative stress appears to be of particular interest as it is common for many toxic effects of OTA (Klauning and Kamendulis, 2004). Reactive oxygen species (ROS) have a significant role in mycotoxicosis, mediating cellular damage (Surai et al. 2008). ROS’s excessive formation leads to oxidative stress, which can trigger cell damage by oxidizing macromolecular structures and modifying their biological functions, ultimately causing cell cycle arrest and cell apoptosis (Ting et al. 2010). The nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor responsible for regulating cellular redox balance in eukaryotic organisms (Habtemariam, 2019). Once Nrf2 is activated, it binds to the antioxidant responsive elements (ARE) in the promoter region of target genes. Hence, activation of Nrf2 leads to the induction of HO-1 expression and other antioxidant proteins (Itoh et al. 2003). Antioxidant treatment is a medical approach for protection against the disturbed oxidant-antioxidant status and has been considered a hopeful remedy for the prevention and treatment of many diseases (Abd El Latif et al., 2021). For that, compounds that ameliorate OTA-induced disorders must be identified to be offered in animal diets and as protective agents for human health. Therefore, substantial efforts have been directed toward identifying natural antioxidants with free radical scavenging action to combat oxidative stress-mediated toxicity. Feed supplements with immunostimulating properties as medicinal plants or probiotics have been widely used within the last years (Abdelhady and El-Abasy 2015; Abdelhady et al. 2017; Markowiak and Śliżewska 2018; Dawood et al. 2020 Moustafa et al. 2020; Assar et al. 2021). Aspergillus awamori has been known as a safe and efficient probiotic 13 microorganism (Lee et al. 2008; Saleh et al. 2015). A. awamori supplementation in animal diet modulates digestive enzymes, therefore enhancing the nutrients digestibility (Tamang et al. 2016), reducing skeletal muscle lipid peroxidation (Saleh et al. 2012 and El-Deep et al. 2014), and improving the immune response of growing rabbits (El-Deep et al. 2021). Ther (...truncated)