Objective To explore the protective effects and underlying mechanisms of H_(2)S against lipid peroxidation-mediated carbonyl stress in the uranium-treated NRK-52E cells.Methods Cell viability was evaluated using CCK-8...Objective To explore the protective effects and underlying mechanisms of H_(2)S against lipid peroxidation-mediated carbonyl stress in the uranium-treated NRK-52E cells.Methods Cell viability was evaluated using CCK-8 assay. Apoptosis was measured using flow cytometry. Reagent kits were used to detect carbonyl stress markers malondialdehyde, 4-hydroxynonenal, thiobarbituric acid reactive substances, and protein carbonylation. Aldehyde-protein adduct formation and alcohol dehydrogenase, aldehyde dehydrogenase 2, aldo-keto reductase, nuclear factor E2-related factor 2(Nrf2), and cystathionine β-synthase(CBS) expression were determined using western blotting or real-time PCR. Sulforaphane(SFP) was used to activate Nrf2. RNA interference was used to inhibit CBS expression.Results GYY4137(an H_(2)S donor) pretreatment significantly reversed the uranium-induced increase in carbonyl stress markers and aldehyde-protein adducts. GYY4137 effectively restored the uraniumdecreased Nrf2 expression, nuclear translocation, and ratio of nuclear to cytoplasmic Nrf2, accompanied by a reversal of the uranium-decreased expression of CBS and aldehyde-metabolizing enzymes. The application of CBS siRNA efficiently abrogated the SFP-enhanced effects on the expression of CBS, Nrf2 activation, nuclear translocation, and ratio of nuclear to cytoplasmic Nrf2 and concomitantly reversed the SFP-enhanced effects of the uranium-induced mRNA expression of aldehyde-metabolizing enzymes.Simultaneously, CBS siRNA reversed the SFP-mediated alleviation of the uranium-induced increase in reactive aldehyde levels, apoptosis rates, and uranium-induced cell viability.Conclusion H_(2)S induces Nrf2 activation and nuclear translocation, which modulates the expression of aldehyde-metabolizing enzymes and the CBS/H_(2)S axis. Simultaneously, the Nrf2-controlled CBS/H_(2)S axis may at least partially promote Nrf2 activation and nuclear translocation. These events form a cycleregulating mode through which H_(2)S attenuates the carbonyl stress-mediated NRK-52E cytotoxicity triggered by uranium.展开更多
基金supported by the National Natural Science Foundation of China(No.82160627)the Natural Science Foundation of the Guangxi Autonomous Region(No.2020GXNFSAA297262)。
文摘Objective To explore the protective effects and underlying mechanisms of H_(2)S against lipid peroxidation-mediated carbonyl stress in the uranium-treated NRK-52E cells.Methods Cell viability was evaluated using CCK-8 assay. Apoptosis was measured using flow cytometry. Reagent kits were used to detect carbonyl stress markers malondialdehyde, 4-hydroxynonenal, thiobarbituric acid reactive substances, and protein carbonylation. Aldehyde-protein adduct formation and alcohol dehydrogenase, aldehyde dehydrogenase 2, aldo-keto reductase, nuclear factor E2-related factor 2(Nrf2), and cystathionine β-synthase(CBS) expression were determined using western blotting or real-time PCR. Sulforaphane(SFP) was used to activate Nrf2. RNA interference was used to inhibit CBS expression.Results GYY4137(an H_(2)S donor) pretreatment significantly reversed the uranium-induced increase in carbonyl stress markers and aldehyde-protein adducts. GYY4137 effectively restored the uraniumdecreased Nrf2 expression, nuclear translocation, and ratio of nuclear to cytoplasmic Nrf2, accompanied by a reversal of the uranium-decreased expression of CBS and aldehyde-metabolizing enzymes. The application of CBS siRNA efficiently abrogated the SFP-enhanced effects on the expression of CBS, Nrf2 activation, nuclear translocation, and ratio of nuclear to cytoplasmic Nrf2 and concomitantly reversed the SFP-enhanced effects of the uranium-induced mRNA expression of aldehyde-metabolizing enzymes.Simultaneously, CBS siRNA reversed the SFP-mediated alleviation of the uranium-induced increase in reactive aldehyde levels, apoptosis rates, and uranium-induced cell viability.Conclusion H_(2)S induces Nrf2 activation and nuclear translocation, which modulates the expression of aldehyde-metabolizing enzymes and the CBS/H_(2)S axis. Simultaneously, the Nrf2-controlled CBS/H_(2)S axis may at least partially promote Nrf2 activation and nuclear translocation. These events form a cycleregulating mode through which H_(2)S attenuates the carbonyl stress-mediated NRK-52E cytotoxicity triggered by uranium.
