E bc k031 s

Reduced glutathione (GSH, µg dL⁻¹) was estimated using GSH colorimetric assay kit (E-BC-K030-S, Elabscience, Houston, TX, USA) according to the method described by Beutler et al. [45 (link)]. Lipid peroxidation was estimated using a malondialdehyde (MDA, nmol mL⁻¹) colorimetric assay kit (E-BC-K025-S, Elabscience, Houston, TX, USA) by measuring thiobarbituric acid reactive substance (TBARS) and expressed in terms of MDA content according to Ohkawa et al. [46 (link)]. MDA, an end product of fatty acid peroxidation, forms a colored complex reacting with Thiobarbituric acid (TBA). The absorbance of the supernatant was measured at 532 nm, and the results were calculated as nmol mL⁻¹. Superoxide dismutase (SOD, U L⁻¹) activity using SOD typed activity assay kit (E-BC-K022-S, Elabscience, Houston, TX, USA) was determined according to Giannopolitis and Ries [47 (link)]. The color reaction was measured at 550 nm, expressed as U L⁻¹. Catalase (CAT, U L⁻¹) activity was determined using a CAT activity assay kit (E-BC-K031-S, Elabscience, Houston, TX, USA) according to the method of Aebi [48 ]. All Oxidative stress markers were determined using a blood chemistry analyzer (HumaLyzer 4000, Human Gesellschaft für Biochemica und Diagnostica mbH, Wiesbaden, Germany).

Reduced glutathione (GSH) concentrations were determined using a GSH colorimetric assay kit (E-BC-K030-S, Elabscience, Texas, USA) in accordance with the protocol established by Beutler et al. [54 (link)]. Using a malondialdehyde (MDA, nmol mL⁻¹) colorimetric test kit (E-BC-K025-S, Elabscience, Texas, USA), we quantified lipid peroxidation in terms of MDA content, as described by Ohkawa et al. [55 (link)]. The activity of superoxide dismutase (SOD, U L⁻¹) was measured using a SOD typed activity assay kit (E-BC-K022-S, Elabscience, Texas, USA) in accordance with Giannopolitis and Ries [56 (link)]; results were expressed as U L⁻¹. The activity of catalase (CAT, U L⁻¹) was measured using a CAT activity assay kit (E-BC-K031-S, Elabscience, Texas, USA) according to the method of Aebi [57 ]. To determine the oxidative stress biomarkers in the kidney tissues, the appropriate weight of kidney tissue was homogenized in PBS (0.01 M, pH 7.4) on ice (1:9, w:v), then tissue homogenates were centrifuged (Sigma, Nussloch, Germany) under cooling at 10,000× g for 10 min. The clear supernatant was collected and preserved, and the GSH, MDA, SOD, and CAT concentrations were determined using the same kits. The blood chemistry analyzer (HumaLyzer 4000, HUMAN Gesellschaft für Biochemica und Diagnostica mbH, Max-Planck-Ring 21, 65205 Wiesbaden, Germany) measured all oxidative stress markers.

Reduced glutathione (GSH, µg dL⁻¹) was estimated using a GSH colorimetric assay kit (E-BC-K030-S, Elabscience, Houston, TX, USA) according to the method described by Beutler et al. [36 (link)]. Lipid peroxidation was assessed using a malondialdehyde (MDA, nmol mL⁻¹) colorimetric assay kit (E-BC-K025-S, Elabscience, Houston, TX, USA) measuring TBARS, and expressed in terms of MDA content according to Ohkawa et al. [37 (link)]. MDA, a byproduct of fatty acid peroxidation, reacts with thiobarbituric acid to form a colored complex (TBA). The absorbance of the supernatant was measured at 532 nm and converted to nmol mL⁻¹. Superoxide dismutase (SOD, U L⁻¹) activity was determined using a SOD activity assay kit (E-BC-K022-S, Elabscience, Houston, TX, USA) according to Giannopolitis and Ries [38 (link)]. At 550 nm, the color reaction was measured and expressed as U L⁻¹. The activity of catalase (CAT, U L⁻¹) was measured using a CAT activity assay kit (E-BC-K031-S, Elabscience, Houston, TX, USA) and the Aebi method [39 ]. A blood chemistry analyzer was used to determine all oxidative stress markers (HumaLyzer 4000, Human Gesellschaft für Biochemica und Diagnostica mbH, Wiesbaden, Germany).

According to the technique outlined by Beutler et al. [43 (link)], reduced glutathione (GSH, g dL⁻¹) was determined using a GSH colorimetric test kit (E-BC-K030-S, Elabscience, Houston, TX, USA). According to Ohkawa et al. [44 (link)], lipid peroxidation was evaluated using a malondialdehyde (MDA, nmol mL⁻¹) colorimetric assay kit (E-BC-K025-S, Elabscience, Houston, TX, USA) by detecting the thiobarbituric acid reactive substance (TBARS) MDA complex. The absorbance of the generated colored complex was measured at 532 nm and calculated as nmol mL⁻¹. Giannopolitis and Ries’ method [45 (link)] was used to measure the activity of superoxide dismutase (SOD, U L⁻¹) using a SOD-type activity assay kit (E-BC-K022-S, Elabscience, Houston, TX, USA). The color reaction was measured at 550 nm, expressed as U L⁻¹. Utilizing a CAT activity test kit (E-BC-K031-S, Elabscience, Houston, TX, USA), the catalase (CAT, U L⁻¹) activity was assessed using the method of Aebi [46 ]. All oxidative stress markers were determined using a blood chemistry analyzer (HumaLyzer 4000, HUMAN Gesellschaft für Biochemica und Diagnostica mbH, Max-Planck-Ring 21, 65205 Wiesbaden, Germany).