Protective effects of curcumin and silymarin against paracetamol induced hepatotoxicity in adult male albino rats

Liu

et al. 2021

fnr

Background: Hypoxia is associated with abnormal cell apoptosis in trophoblast cells, which causes fetal growth restriction and related placental pathologies. Few effective methods for the prevention and treatment of placenta-related diseases exist. Natural products and functional foods have always been a rich source of potential anti-apoptotic drugs. Nobiletin (NOB), a hexamethoxyflavonoid derived from the citrus pomace, shows an anti-apoptotic activity, which is a non-toxic constituent of dietary phytochemicals approved by the Food and Drug Administration. However, their effects on hypoxia-induced human trophoblast cells have not been fully studied. Objective: The aim of this study was to investigate the protective effects of NOB on hypoxia-induced apoptosis of human trophoblast JEG-3 and BeWo cells, and their underlying mechanisms. Design: First, the protective effect of NOB on hypoxia-induced apoptosis of JEG-3 and BeWo cells was studied. Cell viability and membrane integrity were determined by CCK-8 assay and lactate dehydrogenase activity, respectively. Real Time Quantitative PCR (RT-qPCR) and Western blot analysis were used to detect the mRNA and protein levels of HIF1α. Propidium iodide (PI)-labeled flow cytometry was used to detect cell cycle distribution. Cell apoptosis was detected by flow cytometry with Annexin V-FITC and PI double staining, and the expression of apoptosis marker protein cl-PARP was detected by Western blot analysis. Then, the molecular mechanism of NOB against apoptosis was investigated. Computer molecular docking and dynamics were used to simulate the interaction between NOB and p53 protein, and this interaction was verified in vitro by Ultraviolet and visible spectrum (UV-visible spectroscopy), fluorescence spectroscopy and circular dichroism. Furthermore, the changes in the expression of p53 signaling pathway genes and proteins were detected by RT-qPCR and Western blot analysis, respectively. Results: Hypoxia treatment resulted in a decreased cell viability and cell membrane integrity in JEG-3 and BeWo cell lines, and an increased expression of HIF1α, cell cycle arrest in the G1 phase, and massive cell apoptosis, which were alleviated after NOB treatment. Molecular docking and dynamics simulations found that NOB spontaneously bonded to human p53 protein, leading to the change of protein conformation. The intermolecular interaction between NOB and human p53 protein was further confirmed by UV-visible spectroscopy, fluorescence spectroscopy and circular dichroism. After the treatment of 100 μM NOB, a down-regulation of mRNA and protein levels of p53 and p21 and an up-regulation of BCL2/BAX mRNA and protein ratio were observed in JEG-3 cells; however, there was also a down-regulation of mRNA and protein levels observed for p53 and p21 in BeWo cells after the treatment of NOB. The BCL2/BAX ratio of BeWo cells did not change after the treatment of 100 μM NOB. Conclusion: NOB attenuated hypoxia-induced apoptosis in JEG-3 and BeWo cell lines and might be a potential functional ingredient to prevent pregnancy-related diseases caused by hypoxia-induced apoptosis. These findings would also suggest the exploration and utilization of citrus resources, and the development of citrus industry.

Section: Discussionmentioning

confidence: 99%

Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

Liu

et al. 2021

fnr

“…In addition, the heart and kidney sections were incubated with primary antibodies specific for p53 diluted in PBS overnight at 4 °C (mouse monoclonal antibody; Clone: DO-7; Code No. M 7001; diluted 1:25; Dako, CA, USA) [105]. After three washes with PBS, the sections were incubated with a goat anti-rabbit IgG biotin-conjugated secondary antibody (1:2,000; sc‑2040; Santa Cruz Biotechnology, Inc.) at 32 °C for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Investigation of the In-Vivo Cytotoxicity and the In Silico-Prediction of MDM2-p53 Inhibitor Potential of Euphorbia peplus Methanolic Extract in Rats

Abd‐Elhakim

Nassan

Salem

et al. 2019

Toxins

This study explored the probable in vivo cardiac and renal toxicities together with in silico approaches for predicting the apoptogenic potential of Euphorbia peplus methanolic extract (EPME) in rats. Cardiac and renal injury biomarkers were estimated with histopathological and immunohistochemical evaluations of both kidney and heart. The probable underlying mechanism of E. peplus compounds to potentiate p53 activity is examined using Molecular Operating Environment (MOE) docking software and validated experimentally by immunohistochemical localization of p53 protein in the kidney and heart tissues. The gas chromatography/mass spectrometry analysis of E. peplus revealed the presence of nine different compounds dominated by di-(2-ethylhexyl) phthalate (DEHP). Significant elevations of troponin, creatine phosphokinase, creatine kinase–myocardium bound, lactate dehydrogenase, aspartate transaminase, alkaline phosphatase, urea, creatinine, and uric acid were evident in the EPME treated rats. The EPME treated rats showed strong renal and cardiac p53 expression and moderate cardiac TNF-α expression. Further, our in silico results predicted the higher affinity and good inhibition of DEHP, glyceryl linolenate, and lucenin 2 to the MDM2-p53 interface compared to the standard reference 15 a compound. Conclusively, EPME long-term exposure could adversely affect the cardiac and renal tissues probably due to their inflammatory and apoptotic activity. Moreover, the in silico study hypothesizes that EPME inhibits MDM2-mediated degradation of p53 suggesting possible anticancer potentials which confirmed experimental by strong p53 expression in renal and cardiac tissues.

“…Inhibition of liver oxidative stress Pre-clinical model: 50 male C57BL/6 J mice [ 77 ] 11 Silybum marianum (L.) Gaertn Silymarin Lower GSSG content, lower HO-1 induction, alleviated nitrosative stress, decreased p-JNK activation. improve ALT, AST, BUN, SCr and tissue NO levels Pre-clinical model: Male BALB/c mice; Female mice (8–10 weeks, 30–35 g); Male Wistar rats (150–180 g) [ 78 – 80 ] 12 Hypericum monogynum L Hyperoside Inhibition of the formation of toxic intermediates and promotes APAP liver detoxification Pre-clinical model: Male Kunming mice (6 weeks old, 20–25 g) [ 81 ] 13 Garcinia mangostana α-otwistinin Inhibiting oxidative stress and attenuates inflammatory response through NF-κB and MAPK signaling pathways Pre-clinical model: Male ICR mice (4–5 weeks old, 20–25 g) [ 82 ] 14 Glycyrrhiza uralensis Fisch Licochalcone A Protective effect of Nrf2-mediated oxidative stress on APAP-induced liver injury Pre-clinical model: Nrf2 −/− (knockout) C57BL/6 mice [ 83 ] 15 Kaempferol galanga L Kaempferol Antioxidative stress, promoting metabolism and inhibiting inflammation Pre-clinical model: L02 cell [ 84 ] 16 Glycyrrhiza uralensis Fisch Monoammonium glycyrrhizin Regulation of expression of hepatobiliary membrane transporters Pre-clinical model: Male Wistar rats (180–220 g) [ 85 ] …”

Section: Common Drugs and Mechanism Of Dilimentioning

confidence: 99%

Mechanism of drug-induced liver injury and hepatoprotective effects of natural drugs

et al. 2021

Drug-induced liver injury (DILI) is a common adverse drug reaction (ADR) and a serious threat to health that affects disease treatments. At present, no targeted clinical drugs are available for DILI. Traditional natural medicines have been widely used as health products. Some natural medicines exert specific hepatoprotective effects, with few side effects and significant clinical efficacy. Thus, natural medicines may be a promising direction for DILI treatment. In this review, we summarize the current knowledge, common drugs and mechanisms of DILI, as well as the clinical trials of natural drugs and their bioactive components in anticipation of the future development of potential hepatoprotective drugs.