硫化氢对顺铂致犬肾脏氧化损伤及炎症的缓解作用

【目的】 探究硫化氢（H₂S）对顺铂诱导的犬急性肾损伤（AKI）的影响。【方法】 将18只健康成年比格犬随机分为对照组（C）、顺铂组（cis）和硫化氢+顺铂组（H+cis），每组6只。对照组犬经头静脉注射生理盐水，cis组犬经头静脉注射5 mg/kg顺铂，H+cis组犬在注射顺铂前30 min经头静脉注射1 mg/kg NaHS溶液，每隔24 h注射1次，一共3次。注射顺铂72 h后对犬进行麻醉，静脉采血用于检测血清肌酐（Scr）和尿素氮（BUN），HE染色观察各组肾脏病理变化，生化试剂盒检测犬肾脏中谷胱甘肽（GSH）、过氧化氢（H₂O₂）和一氧化氮（NO）含量，实时荧光定量PCR和Western blotting法检测犬肾脏中白介素-1β（IL-1β）、IL-10、肿瘤坏死因子-α（TNF-α）、核因子-κB （NF-κB）、环氧合酶2（COX2）、诱导型一氧化氮合成酶（iNOS）、IL-6、IL-4的相对表达量。【结果】 cis组Scr和BUN水平极显著高于对照组（P<0.01），H+cis组Scr和Bun水平极显著低于cis组（P<0.01）。病理检测结果显示，cis组犬肾脏组织发生明显病理学变化，而H+cis组犬肾脏组织病理变化有所减轻。生化检测结果表明，与C组相比，cis组肾脏中GSH含量极显著下降（P<0.01），H₂O₂和NO含量极显著增加（P<0.01）；与cis组相比，H+cis组犬肾脏中GSH含量极显著上升（P<0.01），而H₂O₂和NO含量极显著下降（P<0.01）。实时荧光定量PCR和Western blotting检测结果表明，与C组相比，cis组促炎因子IL-1β、IL-6、TNF-α、NF-κB和COX2的mRNA和蛋白相对表达量均显著或极显著升高（P<0.05；P<0.01），而抗炎因子IL-4和IL-10的mRNA和蛋白相对表达量均极显著下降（P<0.01）；与cis组相比，H+cis组犬肾脏中抗炎因子IL-4和IL-10的mRNA及蛋白表达量极显著升高（P<0.01），而促炎因子IL-β、IL-6、TNF-α、NF-κB和COX2的mRNA及蛋白相对表达量均极显著下降（P<0.01）。【结论】 H₂S通过提高肾脏抗氧化能力，抑制炎症因子表达来改善顺铂诱导的犬AKI。

Alleviative Effect of Hydrogen Sulfide on Cisplatin-induced Kidney Oxidative Damage and Inflammation in Dogs

【Objective】 The aim of this study was to explore the effect of hydrogen sulfide (H₂S) on the acute kidney injury (AKI) of dog induced by cisplatin.【Method】 18 healthy adult Beagle dogs were randomly divided into three groups:Control group (C), cis group and H+cis group.The dogs in control group were injected with normal saline intravenously, the dogs in cis group were injected with 5 mg/kg cisplatin intravenously, the dogs in H+cis group were injected with 1 mg/kg NaHS intravenously, and after 30 min cisplatin was injected, NaHS solution was injected once every 24 h, a total of 3 times.The dogs were anesthetized 72 h after cisplatin injection, and blood was collected from the vein to test the serum creatinine (Scr) and urea nitrogen (BUN).The kidney pathological changes were observed by HE staining.The contents of glutathione (GSH), hydrogen peroxide (H₂O₂) and nitric oxide (NO) in kidney tissues of dog were detected by biochemical kits.The mRNA and protein expressions of IL-1β, IL-6, TNF-α, NF-κB, COX2, iNOS, IL-4 and IL-10 in kidney were detected by Real-time quantitative PCR and Western blotting.【Result】 The levels of Scr and BUN in cis group were extremely significantly higher than those in control group (P<0.01), and the levels of Scr and BUN in H+cis group were extremely significantly lower than those in cis group (P<0.01).The results of pathological examination showed that there were significant pathological changes in kidney of dogs in cis group, while the pathological changes in kidney of dogs of H+cis group were alleviated.The biochemical test results showed that compared with C group, the content of GSH in kidney of cis group was decreased extremely significantly (P<0.01), and the contents of H₂O₂ and NO were increased extremely significantly (P<0.01);Compared with cis group, the content of GSH in kidney of H+cis group was increased extremely significantly (P<0.01), while the contents of H₂O₂ and NO were decreased extremely significantly (P<0.01).The results of Real-time quantitative PCR and Western blotting showed that compared with C group, the mRNA and protein relative expressions of the proinflammatory factor IL-1β, IL-6, TNF-α, NF-κB and COX2 in cis group were increased significantly or extremely significantly (P<0.05;P<0.01), while the mRNA and protein relative expressions of anti-inflammatory factors IL-4 and IL-10 were decreased extremely significantly (P<0.01);Compared with cis group, the mRNA and protein expressions of anti-inflammatory factors IL-4 and IL-10 in kidney of dogs in H+cis group were extremely significantly higher (P<0.01), while the mRNA and protein relative expressions of pro-inflammatory factor IL-1β, IL-6, TNF-α, NF-κB and COX2 were decreased extremely significantly (P<0.01).【Conclusion】 H₂S could improve cisplatin induced AKI in dogs by enhancing the antioxidant capacity of kindey and inhibiting the expression of inflammatory factors.