硫化氢缓解羰基应激拮抗铀诱导的大鼠肾毒性

Hydrogen sulfide antagonizes uranium-induced rats nephrotoxicity by alleviating carbonyl stress

目的探讨硫化氢(H_(2)S)缓解羰基应激拮抗铀诱导的肾毒性。方法将SD雄性大鼠分成4组:正常对照组、GYY4137(H_(2)S供体)组、铀染毒组和GYY4137治疗组。采用微板法和比色法测定尿液中肌酐与尿素氮含量,HE染色检查肾组织损伤,Tunel法检测肾细胞凋亡,酶联免疫吸附法测定硫代巴比妥酸反应物(TBARS)和4-羟基壬烯醛(4-HNE)含量,硫代巴比妥酸法测定丙二醛(MDA)含量。分光光度法测定蛋白质羰基化(PCO)和8-羟基脱氧鸟苷(8-OhdG)含量,Western blots检测乙醇脱氢酶(ADH1)、乙醛脱氢酶(ALDH2)、醛酮还原酶(AKR7A1)等醛代谢酶的蛋白表达。结果铀染毒后,大鼠肌酐和尿素氮含量降低,活性醛、PCO、8-OhdG的含量增加,出现肾组织结构损伤和肾细胞凋亡,醛代谢酶表达量降低,差异有统计学意义(P<0.05)。GYY4137可缓解铀对肾组织的损伤,降低肾损伤相关指标(P<0.05)。结论H_(2)S通过诱导醛代谢酶表达、促进活性醛代谢和缓解羰基应激发挥拮抗铀产生的大鼠肾毒性。

Objective To investigate whether hydrogen sulfide(H_(2)S)can alleviate uranium-induced rats nephrotoxicity by antagonizing carbonyl stress.Methods SD male rats were divided into four groups:normal control group,GYY4137(H_(2)S donor)treatment group,uranium exposure group,and GYY4137 treatment group.The contents of creatinine and Blood urea nitrogen in urine were determined by microplate method and colorimetry.Kidney tissue damage was examined by HE staining.Kidney cell apoptosis was detected by Tunel method.Thiobarbituric Acid Reactive Substances(TBARS)and 4-Hydroxynonenal(4-HNE)contents were determined by enzyme-linked immunosorbent assay and MDA content was determined by thio Barbituric acid method.The contents of protein carbonylation(PCO)and 8-hydroxy Deoxyguanosine(8-OhdG)were measured by spectrophotometry.Western blots was used to detect the expression of aldehyde metabolic enzymes including alcohol dehydrogenase 1(ADH1),aldehyde dehydrogenase gene 2(ALDH2),aldo-keto-reductase-7A1(AKR7A1).Results Uranium exposure decreased the contents of creatinine and Blood urea nitrogen,led to structural damage of renal tissue,induced kidney cells apoptosis,increased the content of active aldehydes,PCO,8-OhdG,and reduced the expression of aldehyde metabolic enzymes.The difference is statistically significant(P<0.05).However,GYY4137 administration reversed the above experimental indicators of uranium effect(P<0.05).Conclusion H_(2)S can induce the expression of aldehyde metabolism enzymes,promote active aldehyde metabolism,and alleviate carbonyl stress,resulting in antagonize uranium-triggered rats nephrotoxicity.