Journal: Molecular and Cellular Biochemistry

Article Title: Olaparib alleviates diclofenac-induced toxicity in HepG2 cells via modulation of oxidative stress and mitochondrial functions

doi: 10.1007/s11010-026-05581-3

Figure Lengend Snippet: Olaparib diminished diclofenac-induced cell death. A and B Viabilities of HepG2 presented as ratio of dead cells ( A ) and of AML12 ( B ) cells were assessed by trypan blue exclusion ( A ) and MTT ( B ) assays after treating the cells with different combinations of diclofenac (Dic, 200 µM) and olaparib (Ola, 40 µM) for 24 h. Data is given as mean ± SD of three independent experiments running in at least three parallels ( n = 9–16). C and D RTCA assay was used to assess the effect of the drugs on the viability and invasive growth of HepG2 cells. The cells were treated with diclofenac (Dic, 200 µM), olaparib (Ola, 40 µM), NAD + (100 µM) and rotenone (Rot, 1 µM) in the indicated combination for up to 120 h. Cell index calculated from the curves that reflect invasive growth is presented as mean ± SD of three independent experiments running in at least three parallels ( n = 9–12). Calculated p values are indicated above the bars. Differences between the groups were considered significant at p < 0.05. ns, non-significant difference

Article Snippet: Diclofenac and 2’,7’-dichlorodihydrofluorescein diacetate (H 2 DCFDA) were purchased from MedChem Express (Princeton, NJ, USA).

Techniques:

Journal: Molecular and Cellular Biochemistry

Article Title: Olaparib alleviates diclofenac-induced toxicity in HepG2 cells via modulation of oxidative stress and mitochondrial functions

doi: 10.1007/s11010-026-05581-3

Figure Lengend Snippet: Olaparib ameliorated diclofenac induced mitochondrial depolarization. A HepG2 cells were treated with diclofenac (Diclo, 200 µM) and olaparib (Ola, 40 µM) in the indicated combination for 20 h, and live-imaged by fluorescent microscopy after loading with the membrane potential dependent fluorescent dye, JC-1. The results are presented as representative microscopy images of identical fields in the red (aggregated JC-1 indicating polarized mitochondria) and green (monomeric JC-1 indicating depolarized mitochondria) channels. Scale bars in the lower right corner of the merged images represent 25 μm. The accompanying bar diagram presents the average ratio of red to green fluorescence intensity in arbitrary units (a.u.), mean ± SD of three separate experiments running in 2–3 parallels ( n = 6–9). (B) K562 cells were treated with diclofenac (Diclo, 200 µM) and olaparib (Ola, 40 µM) in the indicated combination for 22 h, stained with JC-1 dye, and red and green fluorescence intensities were determined using flow cytometry. Data are presented as red fluorescence in the % of total fluorescence, mean ± SD of three independent experiments ( n = 3). Calculated p values are indicated above the bars. Differences between the groups were considered significant at p < 0.05

Article Snippet: Diclofenac and 2’,7’-dichlorodihydrofluorescein diacetate (H 2 DCFDA) were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Microscopy, Membrane, Fluorescence, Staining, Flow Cytometry

Journal: Molecular and Cellular Biochemistry

Article Title: Olaparib alleviates diclofenac-induced toxicity in HepG2 cells via modulation of oxidative stress and mitochondrial functions

doi: 10.1007/s11010-026-05581-3

Figure Lengend Snippet: Olaparib reduced diclofenac-induced ROS formation. A HepG2 cells were treated with diclofenac (Diclo, 200 µM), olaparib (Ola, 40 µM), or their combination for 24 h, followed by loading the cells with the ROS-sensitive H 2 DCFDA. Fluorescence of 2’,7’-dichlorofluorescein (DCF) resulting from the oxidation of non-fluorescent H 2 DCFDA was assessed using confocal fluorescence microscopy. Data is presented as representative images and as a bar diagram of the DCF fluorescence intensities normalized to a standard and expressed in arbitrary units (a.u.), mean ± SD of three independent experiments ( n = 3). Bright-field images of the same microscopic field are presented underneath the fluorescent images to indicate cell positions. Scale bars in the lower right corner of the fluorescence images represent 50 μm. B Following the same treatment as in ( A ), the cells were fixed, permeabilized, blocked and incubated with anti-GPX4 primary- and anti-mouse secondary antibodies with appropriate wash between steps. Hoechst33342 was used for nuclear counterstaining. Representative immunofluorescence images from three independent experiments ( n = 3) are shown. Scale bars in the lower right corner of the merged images represent 30 μm. C OH − radicals formed in the cell-free Fenton reaction system were measured by photometry. Spontaneous oxidation of salicylic acid was assessed in the absence of H 2 O 2 (Blank), and the reaction was performed in the absence (Control) and presence of 1 mM Olaparib or 1 mM NAC. Absorbance at 510 nm (E 510 ), mean ± SD of three independent experiments ( n = 3) is presented as a bar diagram. D The viability of HepG2 cells was assessed using the trypan blue assay after treating the cells with different combinations of diclofenac (200 µM) and NAC (150 µM or 1 mM) for 24 h. Data is presented as mean ± SD from three independent experiments running in at least three parallels ( n = 10). Calculated p values are indicated above the bars. Differences between the groups were considered significant at p < 0.05. ns, non-significant difference

Article Snippet: Diclofenac and 2’,7’-dichlorodihydrofluorescein diacetate (H 2 DCFDA) were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Fluorescence, Microscopy, Incubation, Immunofluorescence, Control

Journal: Molecular and Cellular Biochemistry

Article Title: Olaparib alleviates diclofenac-induced toxicity in HepG2 cells via modulation of oxidative stress and mitochondrial functions

doi: 10.1007/s11010-026-05581-3

Figure Lengend Snippet: Olaparib diminished diclofenac-induced apoptosis. A Following a 20-hour treatment of HepG2 cells with diclofenac (Diclo, 200 µM), olaparib (Ola, 40 µM), or their combination, the cells were fixed, permeabilized, blocked and incubated with anti-AIF monoclonal antibody and anti-mouse secondary antibody with appropriate washings between the steps. Hoechst33342 was used for nuclear staining, and the cells were visualized under a confocal microscope. The results are presented as representative images and as a bar diagram of the fluorescence intensities normalized to a standard and expressed in arbitrary units (a.u.), mean ± SD of three independent experiments ( n = 3). Scale bars in the lower right corner of the merged images represent 15 μm. Calculated p values are indicated above the bars. Differences between the groups were considered significant at p < 0.05. B The cells were treated as in (A) for 24 h. Subsequently, they were stained with 3% Giemsa solution for 30 min and were imaged using an inverted microscope. Results are presented as representative images from three independent experiments ( n = 3). Arrows indicate nuclear fragmentation and chromatin condensation. C Schematic illustration of the interactions between ROS, PARP inhibition, and nuclear fragmentation. Pointed arrows indicate activation, whereas flat-headed lines indicate inhibition

Article Snippet: Diclofenac and 2’,7’-dichlorodihydrofluorescein diacetate (H 2 DCFDA) were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Incubation, Staining, Microscopy, Fluorescence, Inverted Microscopy, Inhibition, Activation Assay

Journal: Molecular and Cellular Biochemistry

Article Title: Olaparib alleviates diclofenac-induced toxicity in HepG2 cells via modulation of oxidative stress and mitochondrial functions

doi: 10.1007/s11010-026-05581-3

Figure Lengend Snippet: Olaparib attenuated the oxidative phosphorylation-inducing effect of diclofenac in HepG2 cells. A Original plot generated by the Seahorse cellular respirometer from recordings of OCR, after treating the cells with a combination of diclofenac (Diclo, 200 µM) and olaparib (Ola, 40 µM) for 13.5 h. During each recording, oligomycin, FCCP and rotenone plus antimycin A were added at 15, 35, and 55 min of the run (at the arrows). B Original plot of ECAR from the same recordings as for the OCR. (C) OCR/ECAR ratio was calculated from the raw data according to Desousa et al. . All data is presented as mean ± SD, based on three independent experiments ( n = 3). Asterix indicates a significant difference from the control, p < 0.05

Article Snippet: Diclofenac and 2’,7’-dichlorodihydrofluorescein diacetate (H 2 DCFDA) were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Phospho-proteomics, Generated, Control

Journal: Molecular and Cellular Biochemistry

Article Title: Olaparib alleviates diclofenac-induced toxicity in HepG2 cells via modulation of oxidative stress and mitochondrial functions

doi: 10.1007/s11010-026-05581-3

Figure Lengend Snippet: Rotenone enhanced ΔΨm and cell survival in diclofenac-induced toxicity. A HepG2 cells were treated with diclofenac (Diclo, 200 M µM) and rotenone (Rot, 1 µM) for 14 h and then live-imaged by fluorescent microscopy after loading them with the membrane potential dependent fluorescent dye, JC-1. The results are presented as representative microscopic images of identical fields in the red and green channels. Scale bars in the lower right corner of the merged images represent 20 μm. The bar diagram presents the average ratio of red to green fluorescence intensity in arbitrary units (a.u.), mean ± SD of three independent experiments ( n = 3). B and C HepG2 cells were treated with diclofenac (Diclo, 200 µM) and rotenone (Rot, 0.7 or 1 µM) for 24 h before measuring ROS formation by DCF fluorescence ( B ) and cell viability using the MTT assay ( C ). DCF fluorescence and viability are presented as mean ± SD from seven separate experiments ( n = 7). Calculated p values are indicated above the bars. Differences between the groups were considered significant at p < 0.05. ns, non-significant difference; *, significant difference p < 0.05; **, significant difference p < 0.01

Article Snippet: Diclofenac and 2’,7’-dichlorodihydrofluorescein diacetate (H 2 DCFDA) were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Microscopy, Membrane, Fluorescence, MTT Assay

Journal: Food Science & Nutrition

Article Title: Pharmacological Evaluation of Methanolic Fruit Extract of Lannea coromandelica : Antioxidant, Cytotoxic, Antidiabetic, Antibacterial, Anti‐Inflammatory, and Sedative Activities

doi: 10.1002/fsn3.71858

Figure Lengend Snippet: Violin plot and time‐course analysis of paw‐edema inhibition showing dose‐dependent anti‐inflammatory activity of ME compared with diclofenac in mice.

Article Snippet: Square Pharmaceuticals Limited is a Bangladeshi pharmaceutical company that supplies diclofenac sodium and loperamide.

Techniques: Inhibition, Activity Assay