Discipline of Medical Biochemistry and Chemical Pathology, Faculty of Health Sciences, George Campbell Building, Howard College, University of KwaZulu-Natal, Durban, 4041, South Africa.
Discipline of Medical Biochemistry and Chemical Pathology, Faculty of Health Sciences, George Campbell Building, Howard College, University of KwaZulu-Natal, Durban, 4041, South Africa; Department of Human Biology, Medical School, Nelson Mandela University, Port Elizabeth, South Africa.
Toxicon. 2022 Apr 30;210:58-65. doi: 10.1016/j.toxicon.2022.02.018. Epub 2022 Feb 22.
Patulin (PAT) is a mycotoxin produced by Penicillium and other fungi that contaminate fruit. PAT targets the kidney and is associated with nephrotoxicity. Micro-RNAs (miRNA) may offer new insights into PAT-induced nephrotoxicity. Cytochrome P family 1, subfamily B, polypeptide 1 (CYP1B1), involved in metabolism of dietary toxins is negatively regulated by miR-27b and linked with the nuclear factor kappa B (NF-κB) pathway and peroxisome proliferator activated receptor gamma (PPARɣ) in renal fibrosis. This study investigated the effects of PAT on miR-27b, CYP1B1, PPARɣ and cytotoxicity in human kidney (HEK293) cells. HEK293 cells were exposed to PAT (2.5 μM, 24h). Protein expression of CYP1B1, PPARɣ, NF-κB (p65), pNF-κB (p65) (phospho-Ser563) and cleaved PARP-1 was quantified using western blotting. QPCR evaluated mRNA levels of CYP1B1, IL-6, miR-27b, OGG1, mtDNA, TFAM and UCP2. Mitochondrial membrane potential and phosphatidylserine (PS) externalization was evaluated by flow cytometry while levels of ATP and caspase -9, -8, -3/7 activity was measured using luminometry. PAT significantly decreased miR-27b levels (p = 0.0014) and increased CYP1B1 mRNA (p = 0.0015) and protein (p = 0.0013) levels. PPARɣ protein expression was significantly increased (p = 0.0002) and associated with decreased NF-κB activation (p = 0.0273) and IL-6 mRNA levels (p = 0.0265). Finally, PAT significantly compromised mitochondrial repair mechanisms and increased apoptotic biomarkers. PAT altered miR-27b levels and PPARɣ, with associated changes to NF-κB activation, downstream IL-6 and CYP1B1 expression. These results show that PAT impairs detoxification mechanisms leading to mitochondrial damage and apoptosis. In conclusion, PAT altered the epigenetic environment and impaired detoxification processes, supporting a mechanism for nephrotoxic outcomes.
棒曲霉素（PAT）是一种由青霉和其他真菌产生的真菌毒素，会污染水果。PAT 靶向肾脏并与肾毒性有关。Micro-RNAs（miRNA）可能为 PAT 诱导的肾毒性提供新的见解。细胞色素 P 家族 1，亚家族 B，多肽 1（CYP1B1）参与膳食毒素的代谢，受 miR-27b 负调控，并与核因子 kappa B（NF-κB）途径和过氧化物酶体增殖物激活受体 γ（PPARγ）在肾纤维化中相关联。本研究探讨了 PAT 对人肾（HEK293）细胞中 miR-27b、CYP1B1、PPARγ和细胞毒性的影响。HEK293 细胞暴露于 PAT（2.5 μM，24 小时）。使用 Western blot 定量测定 CYP1B1、PPARγ、NF-κB（p65）、pNF-κB（p65）（磷酸化-Ser563）和裂解 PARP-1 的蛋白表达。QPCR 评估 CYP1B1、IL-6、miR-27b、OGG1、mtDNA、TFAM 和 UCP2 的 mRNA 水平。通过流式细胞术评估线粒体膜电位和磷脂酰丝氨酸（PS）外翻，使用发光法测量 ATP 和 caspase-9、-8、-3/7 活性。PAT 显著降低 miR-27b 水平（p=0.0014）并增加 CYP1B1 mRNA（p=0.0015）和蛋白（p=0.0013）水平。PPARγ 蛋白表达显著增加（p=0.0002），与 NF-κB 激活减少（p=0.0273）和 IL-6 mRNA 水平降低相关（p=0.0265）。最后，PAT 显著损害了线粒体修复机制并增加了凋亡生物标志物。PAT 改变了 miR-27b 水平和 PPARγ，随之改变 NF-κB 激活、下游 IL-6 和 CYP1B1 表达。这些结果表明，PAT 损害了解毒机制，导致线粒体损伤和细胞凋亡。总之，PAT 改变了表观遗传环境并损害了解毒过程，支持了肾毒性结果的一种机制。
