Amelioration of cyclophosphamide-induced DNA damage, oxidative stress, and hepato- and neurotoxicity by Piper longum extract in rats: The role of γH2AX and 8-OHdG

Abstract

Background: The identification of genoprotectants is a promising strategy for improving human health. Piper longum has drawn scientific attention because of its diverse biological effects and traditional utilization. The current investigation aims to evaluate the genome-stabilizing potential of Piper longum against cyclophosphamide-associated genotoxicity.Methods: We adopted a funnel screening with a three-tier evaluation approach, where Piper longum was investigated in an acellular medium, peripheral blood lymphocytes, and a rodent model. The genoprotective action of the Piper longum extract was initially performed with plasmid pBluescript SK(-) DNA. Furthermore, the extract and various fractions were screened against cyclophosphamide-induced genotoxicity using a cytokinesis-block micronucleus assay and a chromosomal aberration assay in human peripheral blood lymphocytes. The genome-stabilizing action of the extract and potent (hexane) fraction was further confirmed in vivo in Wistar albino rats by evaluating them using mammalian erythrocyte micronucleus tests, DNA fragmentation, oxidative stress markers, 8-hydroxy-2-deoxyguanosine (8-OHdG), γH2AX, and histopathological lesions in the liver and hippocampus. Additionally, acute and sub-acute toxicity studies were conducted following the Organization for Economic Co-operation and Development (OECD) guidelines for rats. Furthermore, the extract was quantified and characterized by high-performance thin-layer chromatography (HPTLC), ultra-high performance liquid chromatography–mass spectroscopy (UPLC-MS), and gas chromatography–mass spectrometry (GC-MS).Results: The Piper longum ethanol extract was shown to protect plasmid pBluescript SK(-) DNA against H2O2-induced strand breaks. In human lymphocytes, the extract and hexane fraction showed a reduction in micronucleus formation (p < 0.001) and chromosomal aberrations (p < 0.01) against cyclophosphamide. Furthermore, the extract and fraction treatment, when administered at 200 mg/kg for 28 days in Wistar rats, restored cyclophosphamide-induced genomic instability by reducing micronucleus formation and DNA fragmentation; restoring redox homeostasis; decreasing 8-OHdG, a hallmark of oxidative DNA damage; reducing γH2AX, a DNA double-strand break (DSB) marker; and preserving the liver and hippocampus against histopathological lesions. The extract and fraction revealed no signs of systemic toxicity at the used doses. Piperine and piperlongumine are the major alkaloids quantified along with the presence of flavonoids in the ethanol extract and the presence of fatty acids and terpenoids in the hexane fraction of Piper longum.Conclusion: Our investigation confirms the genoprotective action of Piper longum by reducing cyclophosphamide-associated cytogenotoxicity, oxidative stress, hepato- and neurotoxicity, oxidative DNA damage, and DNA double-strand breaks. The outcomes are critical for mitigating the genotoxic effects of chemotherapy recipients, requiring further attention.

Temperature-Dependent Bioaccumulation, Metabolism, and Hepatotoxicity of Flufiprole in Lizards (Eremias argus)

“…As higher flufiprole metabolite concentrations were detected at 35 °C than that at 15 or 25 °C, it can be predicted that the more serious liver lesions in the higher-temperature group were induced by the flufiprole oxidative reaction during metabolism. The inflammation in the liver caused by flufiprole may have a protective effect on the liver . All these lesions, including inflammatory cell infiltration and porphyrin pigment deposition, were also highly related to oxidative stress.…”

“…The inflammation in the liver caused by flufiprole may have a protective effect on the liver. 54 All these lesions, including inflammatory cell infiltration and porphyrin pigment deposition, were also highly related to oxidative stress. The oxidative DNA damage induced by diazinon also showed serious inflammation in rabbit liver.…”

Neratinib stimulates senescence of mammary cancer cells by reducing the levels of SIRT1

“…8-OHdG and γH2AX are critical biomarkers of DNA damage [ 17 ]. AU565 cells were stimulated with Neratinib (1, 2.5, and 5 μM) for 72 h. The 8-OHdG ( Figure 3A ) and γH2AX ( Figure 3B ) levels in AU565 cells were remarkably elevated as the concentration of Neratinib increased from 1 to 5 μM, implying a facilitating effect of Neratinib on DNA damage in AU565 cells.…”

Molecular mechanisms underlying cyclophosphamide-induced cognitive impairment and strategies for neuroprotection in preclinical models

“…In addition, various studies demonstrated that oxidative stress elicited by cyclophosphamide in the brain is mediated through overproduction of reactive oxygen species (ROS), GSH depletion and inhibition of antioxidant enzymes activities [13,14]. In addition, it was found that the activation of angiotensin converting enzyme and the resultant excessive production of angiotensin II by cyclophosphamide has been implicated in ROS generation and hence the subsequent oxidative brain injury [15].The massive generation of ROS prompts oxidative damage to numerous cellular components resulting in lipid peroxidation, DNA damage and mitochondrial dysfunction [14,[16][17][18]. Mechanisms by which cyclophosphamide-induced oxidative stress is shown in Fig.…”

“…Notably, nerolidol has exhibited worthy neuroprotective activity [87,88]. Iqbual et al [18] has examined the effect of nerolidol on cyclophosphamide-induced chemobrain. In this study, mice were given nerolidol orally at doses of 200 or 400 mg/kg for 14 days then cyclophosphamide (200 mg/ kg i.p.)…”