Objective: To investigate the effects of cadmium (Cd) on antioxidant enzymes in testis of mice and the protective effect of vitamin C (VC). Methods: A total of 72 male Kunming mice of clean grade were divided into four groups (n=18): the control group, the Cd group (CdCl₂ 3 mg/kg), the VC group (200 mg/kg), and the VC (200 mg/kg) +Cd group (CdCl₂ 3 mg/kg). Mice were poisoned once a day, exposed for 1 and 3 days and were treated with VC at the same time. Twenty-four hours after exposure on the 1st and 3rd day, half of the mice in each group were weighed, the serum and testis tissues were collected. Testicular organ coefficient, malondialdehyde (MDA) and superoxide dismutase (SOD) in serum and testis tissues, and glutathione peroxidase (GSH-Px), reduced glutathione (GSH), oxidized glutathione (GSSG) and total glutathione (T-GSH) in testis tissues were detected. Results: Compared with the control group, the body weight and testicle organ coefficient of mice in the Cd group were decreased on the 1st and 3rd day; after 3 days of exposure, the serum SOD in the Cd group was decreased significantly and MDA was increased significantly (P＜0.05); the levels of SOD, GSH-Px, T-GSH and GSH/GSSG of testis in the Cd group were increased significantly on the 1st day (P＜0.05), while all the above indexes were decreased significantly on the 3rd day (P＜0.05), and the content of MDA was increased significantly on the 1st and 3rd days in the Cd group (P＜0.05); after VC treatment, the degree of reduction was decreased. Compared with the Cd group, the serum SOD and MDA levels in the VC+ Cd group were significantly different after 3 days of exposure (P＜0.05); the changes of SOD, GSH-Px, T-GSH and GSH/GSSG levels of the testis in the VC+ Cd group were significantly different on the 1st and 3rd day of exposure (P＜0.05), and the MDA level of the testis in the VC+ Cd group was decreased significantly on the 3rd day of exposure (P＜0.05). Compared with the Cd group for 1 day, the level of serum SOD exposed for 3 days was decreased significantly (P＜0.05), and the changes of testis indexes were also significantly different (P＜0.05). Conclusion: VC treatment can improve the antioxidant function of cadmium-loaded mice to some extent, and has protective effect on oxidative damage of testis.
Animals ; Antioxidants/pharmacology* ; Ascorbic Acid/pharmacology* ; Cadmium/toxicity* ; Glutathione ; Glutathione Disulfide/pharmacology* ; Glutathione Peroxidase ; Male ; Superoxide Dismutase ; Testis

OBJECTIVETo investigate the alterations in auditory brainstem evoked responses (ABRs) and the changes of carboplatin-induced ototoxicity in the cochlear oxidant/antioxidant systems and otoprotection by an antioxidant lipoate.

METHODSMale wistar rats were divided into four groups and treated as follows: 1) vehicle (saline) control, 2) carboplatin (256 mg/kg, i.p.), 3) lipoate (100 mg/kg, i.p.), 4) lipoate + carboplatin. Post-treatment ABRs were performed after four days and rats were sacrificed with their cochleae harvested and analyzed.

RESULTSCarboplatin significantly elevated ABR threshold above the pretreatment thresholds. Lipoate+carboplatin treated rats showed decreased elevation of hearing threshold. Carboplatin significantly depleted cochlear reduced to oxizized glutathione (GSH/GSSG) ratio, whereas lipoate+carboplatin treatment increased GSH/GSSG ratio. Carboplatin significantly decreased cochlear copper zinc-superoxide dismutase (CuZn-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and glutathione-S-transferase (GST) activities and enzyme protein expressions and a significant increase in Mn-SOD activity, protein expression and malondialdehyde (MDA) level. Cochlear antioxidant enzyme activities, enzyme protein expressions and MDA level were partially restored in lipoate+carboplatin treated rats, compared to carboplatin alone.

CONCLUSIONCarboplatin-induced ototoxicity is related to impairment of cochlear antioxidant system and otoprotection conferred by lipoate is associated with partial sparing of the cochlear antioxidant defense system.

Animals ; Antioxidants ; pharmacology ; Auditory Threshold ; drug effects ; Carboplatin ; Catalase ; metabolism ; Cochlea ; drug effects ; enzymology ; metabolism ; Evoked Potentials, Auditory, Brain Stem ; drug effects ; Glutathione ; metabolism ; Glutathione Disulfide ; metabolism ; Glutathione Peroxidase ; metabolism ; Glutathione Reductase ; metabolism ; Glutathione Transferase ; metabolism ; Hearing Loss, Sensorineural ; chemically induced ; Lipid Peroxidation ; Male ; Malondialdehyde ; metabolism ; Protective Agents ; pharmacology ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism ; Thioctic Acid ; pharmacology

OBJECTIVE: To explore the role of Runt-related transcription factor 3 (RUNX3) in metabolic regulation of trastuzumab-resistant gastric cancer cells and investigate the mechanism of RUNX3 knockdown-mediated reversal of trastuzumab resistance. METHODS: We performed a metabolomic analysis of trastuzumab-resistant gastric cancer cells (NCI N87R) and RUNX3 knockdown cells (NCI N87R/RUNX3) using ultra performance liquid chromatography (UPLC) coupled with Q Exactive Focus Orbitrap mass spectrometry (MS). Multivariate combined with univariate analyses and MS/MS ion spectrums were used to screen the differential variables. MetaboAnalyst 5.0 database was employed for pathway enrichment analysis. Differential metabolites-genes regulatory relationships were constructed based on OmicsNet database. The changes in GSH/GSSG and NADPH/NADP ratios in NCI N87R/RUNX3 cells were measured using detection kits. RESULTS: The metabolic profile of NCI N87R cells was significantly altered after RUNX3 knockdown, with 81 differential metabolites identified to contribute significantly to the classification, among which 43 metabolites were increased and 38 were decreased (P < 0.01). In NCI N87R cells, RUNX3 knockdown resulted in noticeable alterations in 8 pathways involving glutamine metabolism, glycolysis, glycerophospholipid, nicotinate-nicotinamide and glutathione metabolism, causing also significant reduction of intracellular GSH/GSSG and NADPH/NADP ratios (P < 0.01). The differential metabolites-genes network revealed a regulatory relationship between the metabolic molecules and genes. CONCLUSION RUNX3 reverses trastuzumab resistance in gastric cancer cells by regulating energy metabolism and oxidation-reduction homeostasis and may serve as a potential therapeutic target for trastuzumab-resistant gastric cancer.
Chromatography, High Pressure Liquid ; Core Binding Factor Alpha 3 Subunit/genetics* ; Glutathione Disulfide ; Humans ; Metabolomics ; NADP ; Stomach Neoplasms/genetics* ; Tandem Mass Spectrometry ; Trastuzumab/pharmacology*

BACKGROUND: Gallbladder cancer (GBC) is the most common malignant tumor of biliary tract. Isoliquiritigenin (ISL) is a natural compound with chalcone structure extracted from the roots of licorice and other plants. Relevant studies have shown that ISL has a strong anti-tumor ability in various types of tumors. However, the research of ISL against GBC has not been reported, which needs to be further investigated. METHODS: The effects of ISL against GBC cells in vitro and in vivo were characterized by cytotoxicity test, RNA-sequencing, quantitative real-time polymerase chain reaction, reactive oxygen species (ROS) detection, lipid peroxidation detection, ferrous ion detection, glutathione disulphide/glutathione (GSSG/GSH) detection, lentivirus transfection, nude mice tumorigenesis experiment and immunohistochemistry. RESULTS: ISL significantly inhibited the proliferation of GBC cells in vitro . The results of transcriptome sequencing and bioinformatics analysis showed that ferroptosis was the main pathway of ISL inhibiting the proliferation of GBC, and HMOX1 and GPX4 were the key molecules of ISL-induced ferroptosis. Knockdown of HMOX1 or overexpression of GPX4 can reduce the sensitivity of GBC cells to ISL-induced ferroptosis and significantly restore the viability of GBC cells. Moreover, ISL significantly reversed the iron content, ROS level, lipid peroxidation level and GSSG/GSH ratio of GBC cells. Finally, ISL significantly inhibited the growth of GBC in vivo and regulated the ferroptosis of GBC by mediating HMOX1 and GPX4 . CONCLUSION ISL induced ferroptosis in GBC mainly by activating p62-Keap1-Nrf2-HMOX1 signaling pathway and down-regulating GPX4 in vitro and in vivo . This evidence may provide a new direction for the treatment of GBC.
Animals ; Mice ; Carcinoma in Situ ; Chalcones/pharmacology* ; Ferroptosis ; Gallbladder Neoplasms/genetics* ; Glutathione Disulfide ; Kelch-Like ECH-Associated Protein 1 ; Mice, Nude ; NF-E2-Related Factor 2/genetics* ; Reactive Oxygen Species ; Humans

OBJECTIVETo study the effects of Astragaloside (AST) and Astragalus Saponin I (ASI) on metabolism of free radical and immune function in senescent rats treated by HC.

METHODHydrocorisone (HC) was used to estabilsh the aging model in rats. The content of molondialdehyde (MDA), glutathoine (GSH) and oxidized glutathoine (GSSG) in liver and brain was detected according to kit. The activity of Mn superoride dismulase (Mn-SOD) and catalase (CAT) was also surveyed by kit. Concanavalin (ConA) was used to detect the proliferation and interleukin-2 (IL-2) production of splenocytes.

RESULTCompared with HC control, AST and ASI could decrease the content of MDA, GSH and GSSG in liver and brain, increase the activity of Mn-SOD and CAT, and promote the proliferation and interleukin-2 (IL-2) activity of splenocytes.

CONCLUSIONAST and ASI could delay the aging effect in rats treated by HC, and its mechanism maybe the antioxidant and regulating immunity.

Aging ; drug effects ; metabolism ; Animals ; Astragalus membranaceus ; chemistry ; Catalase ; metabolism ; Cell Proliferation ; drug effects ; Cerebral Cortex ; metabolism ; Female ; Free Radicals ; metabolism ; Glutathione ; metabolism ; Glutathione Disulfide ; metabolism ; Hydrocortisone ; toxicity ; Interleukin-2 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saponins ; isolation & purification ; pharmacology ; Spleen ; cytology ; Superoxide Dismutase ; metabolism ; Triterpenes ; isolation & purification ; pharmacology

OBJECTIVETo investigate the in vitro protective effect of Pinus massoniana bark extracts (PMBE) against cisplatin-induced nephrotoxicity in human embryonic kidney cells (HEK293), and preliminarily study its mechanism.

METHODHuman embryonic kidney cells (HEK293) were cultured in vitro. The MTT assay was adopted to test the effect of PMBE and cisplatin on growth of HEK293 cells, and the protective effect of PMBE on cisplatin-induced nephrotoxicity of HEK293, and then detect the intracellular reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH) content, catalase (CAT), superoxide dismutase (SOD) and activity of thioredoxin reductase (TrxR).

RESULTPMBE could promote growth of HEK293 cells at low concentrations, but generate slight nephrotoxicity at high concentration. Cisplatin could inhibit growth of HEK293 cells, increase ROS and MDA content, while reducing SOD, CAT and TrxR. The pre-protective PMBE was added to reduce cisplatin's injury to HEK293 cells, ROS, MDA and GSH content, SOD, CAT and TrxR within certain range.

CONCLUSIONPMBE at specific concentration has the protective effect in cisplatin-induced nephrotoxicity in HEK293 cells. Its mechanism may be related to PMBE's antioxidant activity.

Animals ; Antioxidants ; metabolism ; Cisplatin ; toxicity ; Epithelial Cells ; drug effects ; enzymology ; metabolism ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; HEK293 Cells ; Humans ; Kidney ; drug effects ; enzymology ; metabolism ; Malondialdehyde ; metabolism ; Mice ; Pinus ; chemistry ; Plant Bark ; chemistry ; Plant Extracts ; pharmacology ; Protective Agents ; pharmacology ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Thioredoxin-Disulfide Reductase ; metabolism

Oxidative stress plays a crucial role in cadmium (Cd)-induced myocardial injury. Mitsugumin 53 (MG53) and its mediated reperfusion injury salvage kinase (RISK) pathway have been demonstrated to be closely related to myocardial oxidative damage. Potentilla anserina L. polysaccharide (PAP) is a polysaccharide with antioxidant capacity, which exerts protective effect on Cd-induced damage. However, it remains unknown whether PAP can prevent and treat Cd-induced cardiomyocyte damages. The present study was desgined to explore the effect of PAP on Cd-induced damage in H9c2 cells based on MG53 and the mediated RISK pathway. For in vitro evaluation, cell viability and apoptosis rate were analyzed by CCK-8 assay and flow cytometry, respectively. Furthermore, oxidative stress was assessed by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining and using superoxide dismutase (SOD), catalase (CAT), and glutathione/oxidized glutathione (GSH/GSSG) kits. The mitochondrial function was measured by JC-10 staining and ATP detection assay. Western blot was performed to detect the expression of proteins related to MG53, the RISK pathway, and apoptosis. The results indicated that Cd increased the levels of reactive oxygen species (ROS) in H9c2 cells. Cd decreased the activities of SOD and CAT and the ratio of GSH/GSSG, resulting in decreases in cell viability and increases in apoptosis. Interestingly, PAP reversed Cd-induced oxidative stress and cell apoptosis. Meanwhile, Cd reduced the expression of MG53 in H9c2 cells and inhibited the RISK pathway, which was mediated by decreasing the ratio of p-Akt^Ser473/Akt, p-GSK3β^Ser9/GSK3β and p-ERK1/2/ERK1/2. In addition, Cd impaired mitochondrial function, which involved a reduction in ATP content and mitochondrial membrane potential (MMP), and an increase in the ratio of Bax/Bcl-2, cytoplasmic cytochrome c/mitochondrial cytochrome c, and Cleaved-Caspase 3/Pro-Caspase 3. Importantly, PAP alleviated Cd-induced MG53 reduction, activated the RISK pathway, and reduced mitochondrial damage. Interestingly, knockdown of MG53 or inhibition of the RISK pathway attenuated the protective effect of PAP in Cd-induced H9c2 cells. In sum, PAP reduces Cd-induced damage in H9c2 cells, which is mediated by increasing MG53 expression and activating the RISK pathway.
Cadmium/metabolism* ; Caspase 3/metabolism* ; Potentilla/metabolism* ; Glycogen Synthase Kinase 3 beta/pharmacology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Cytochromes c/metabolism* ; Glutathione Disulfide/pharmacology* ; Oxidative Stress ; Myocytes, Cardiac ; Reactive Oxygen Species/metabolism* ; Reperfusion Injury/metabolism* ; Apoptosis ; Polysaccharides/pharmacology* ; Adenosine Triphosphate/metabolism*