Department of Biochemistry, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey.
Department of Pathology, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey.
Environ Toxicol. 2021 Aug;36(8):1600-1617. doi: 10.1002/tox.23156. Epub 2021 Apr 28.
In this study, we investigated the effects of hesperidin (HSP) on oxidants/antioxidants status, inflammation, apoptotic, and autophagic activity in hepato-renal toxicity induced by chronic chlorpyrifos (CPF) exposure in rats. We used a total of 35 male albino rats in five groups of seven: control, HSP 100, CPF, CPF + HSP50, and CPF + HSP100. After rats were sacrificed, blood, liver, and kidney samples were collected. Serum levels of aspartate aminotransferases (ALT and AST), alkaline phosphatase (ALP), creatinine, and urea were tested. Then, contents of the superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), glutathione peroxidase (GPx), and glutathione (GSH) were measured to detect the level of oxidative stress in rat liver and renal tissues. We measured inflammatory and autophagy markers of chlorpyrifos induced oxidative stress in the liver and kidney tissues including TNF-α, iNOS, IL-1 β, COX-2, NF-κB, MAPK14, and Beclin-1 using ELISA. Histopathological findings were also examined followed by immunohistochemical determination of 8-OHdG expression. Real-time PCR (RT-PCR) was used to examine Cas-3, Bax, Bcl-2, PARP-1, and VEGF, which are associated with apoptosis, autophagy, DNA, and endothelial damage, respectively. In addition, PARP-1 activity was supported by western blot and immunofluorescence, VEGF activity was supported by western blot methods. Treatment with HSP reduced the effect of CPF on ALT, AST, ALP, and total proteins, and increased its effect on tissue antioxidants. PARP/VEGF, apoptotic, pro-apoptotic, anti-apoptotic, and autophagic gene expressions were regulated, and Caspase-3 and Bax expressions were decreased; Bcl-2 expression increased in both the liver and kidney samples, and positivity of 8-OHdG and PARP-1 were reduced in the CPF plus HSP-treated group. Overall, the study demonstrates that HSP may reduce the effects of hepato-renal toxicity caused by CPF by regulating oxidative stress, inflammation, apoptosis, autophagy, and PARP/VEGF genes at biochemical, cellular, and molecular levels.
在这项研究中，我们调查了橙皮苷（HSP）对慢性毒死蜱（CPF）暴露引起的肝肾功能毒性中氧化剂/抗氧化剂状态、炎症、凋亡和自噬活性的影响。我们总共使用了 35 只雄性白化大鼠，分为五组，每组 7 只：对照组、HSP100 组、CPF 组、CPF+HSP50 组和 CPF+HSP100 组。大鼠死后，采集血液、肝脏和肾脏样本。检测血清中天冬氨酸转氨酶（ALT 和 AST）、碱性磷酸酶（ALP）、肌酐和尿素水平。然后，测量超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、丙二醛（MDA）、谷胱甘肽过氧化物酶（GPx）和谷胱甘肽（GSH）的含量，以检测大鼠肝和肾组织中的氧化应激水平。我们使用 ELISA 测量了氯吡硫磷诱导的肝和肾组织氧化应激中的炎症和自噬标志物，包括 TNF-α、iNOS、IL-1β、COX-2、NF-κB、MAPK14 和 Beclin-1。还检查了组织病理学发现，并通过免疫组化测定 8-OHdG 表达进行了免疫组化检测。实时 PCR（RT-PCR）用于检测 Cas-3、Bax、Bcl-2、PARP-1 和 VEGF，它们分别与凋亡、自噬、DNA 和内皮损伤有关。此外，通过 Western blot 和免疫荧光法支持 PARP-1 活性，通过 Western blot 方法支持 VEGF 活性。HSP 处理减轻了 CPF 对 ALT、AST、ALP 和总蛋白的影响，增加了其对组织抗氧化剂的影响。PARP/VEGF、凋亡、促凋亡、抗凋亡和自噬基因表达得到调节，Caspase-3 和 Bax 表达减少；肝和肾组织中 Bcl-2 表达增加，CPF 加 HSP 治疗组中 8-OHdG 和 PARP-1 的阳性率降低。总的来说，该研究表明，HSP 可能通过在生化、细胞和分子水平上调节氧化应激、炎症、凋亡、自噬和 PARP/VEGF 基因来减轻 CPF 引起的肝肾功能毒性。
