硫化氢对顺铂致犬肾脏氧化损伤及炎症的缓解作用

doi:10.16431/j.cnki.1671-7236.2022.06.037

摘要/Abstract

摘要： 【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。

关键词: 犬; 硫化氢; 急性肾损伤; 炎症; 氧化应激

Abstract: 【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.

Key words: dogs; hydrogen sulfide; acute kidney injury; inflammation; oxidative stress

中图分类号:

S852.2

王爽, 刘星尧, 贾昊天, 刘云. 硫化氢对顺铂致犬肾脏氧化损伤及炎症的缓解作用[J]. 中国畜牧兽医, 2022, 49(6): 2376-2384.

WANG Shuang, LIU Xingyao, JIA Haotian, LIU Yun. Alleviative Effect of Hydrogen Sulfide on Cisplatin-induced Kidney Oxidative Damage and Inflammation in Dogs[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2376-2384.

参考文献

[1] VANMASSENHOVE J, KIELSTEIN J, JORRES A, et al.Management of patients at risk of acute kidney injury[J].Lancet, 2017, 389(10084):2139-2151.
[2] YAO W, CHEN Y, LI Z, et al.Single cell RNA sequencing identifies a unique inflammatory macrophage subset as a druggable target for alleviating acute kidney injury[J].Advanced Science (Weinh), 2022, 3:e2103675.
[3] ZHENG X, DENG F, SHARMA I, et al.Myo-inositol oxygenase overexpression exacerbates cadmium-induced kidney injury via oxidant stress and necroptosis[J].American Journal of Physiology Renal Physiology, 2022, 322(3):F344-F359.
[4] PABLA N, DONG Z.Cisplatin nephrotoxicity:Mechanisms and renoprotective strategies[J].Kidney International, 2008, 73(9):994-1007.
[5] OZKOK A, EDELSTEIN C L.Pathophysiology of cisplatin-induced acute kidney injury[J].BioMed Research International, 2014, 2014:967826.
[6] 刘建民, 陈晓农, 江永娣.急性硫化氢中毒致心肌损伤一例[J].中华内科杂志, 1995, 4:286. LIU J M, CHEN X N, JIANG Y D.A case of myocardial injury caused by acute hydrogen sulfide poisoning[J].Chinese Journal of Internal Medicine, 1995, 4:286.(in Chinese)
[7] 张劲松, 蒋蕾, 张华忠.上皮钠通道在硫化氢气体引起急性肺水肿中的作用[A].中华医学会灾难医学分会[C].2015. ZHANG J S, JIANG L, ZHANG H Z.Role of epithelial sodium channel in acute pulmonary edema induced by hydrogen sulfide gas[A].Disaster Medicine Branch of Chinese Medical Association[C].2015.(in Chinese)
[8] ABE K, KIMURA H.The possible role of hydrogen sulfide as an endogenous neuromodulator[J].Journal of Neuroscience, 1996, 16(3):1066-1071.
[9] KIMURA Y, GOTO Y, KIMURA H.Hydrogen sulfide increases glutathione production and suppresses oxidative stress in mitochondria[J].Antioxidants & Redox Signaling, 2010, 12(1):1-13.
[10] TAMIZHSELVI R, SUN J, KOH Y H, et al.Effect of hydrogen sulfide on the phosphatidylinositol 3-kinase-protein kinase B pathway and on caerulein-induced cytokine production in isolated mouse pancreatic acinar cells[J].The Journal of Pharmacology and Experimental Therapeutics, 2009, 329(3):1166-1177.
[11] CHEN Y H, WU R, GENG B, et al.Endogenous hydrogen sulfide reduces airway inflammation and remodeling in a rat model of asthma[J].Cytokine, 2009, 45(2):117-123.
[12] LIN Y, LIN J Q, GUO Y, et al.Atorvastatin protects against contrast-induced acute kidney injury via upregulation of endogenous hydrogen sulfide[J].Renal Failure, 2020, 42(1):270-281.
[13] DAUGAARD G, ABILDGAARD U, HOLSTEIN-RATHLOU N H, et al.Effect of cisplatin on renal haemodynamics and tubular function in the dog kidney[J].International Journal of Andrology, 1987, 10(1):347-351.
[14] LING K, XU A, CHEN Y, et al.Protective effect of a hydrogen sulfide donor on balloon injury-induced restenosis via the Nrf2/HIF-1α signaling pathway[J].International Journal of Molecular Medicine, 2019, 43(3):1299-1310.
[15] 崔永华, 汪洋, 虞正权.硫化氢对大鼠蛛网膜下腔出血后早期脑损伤的保护作用[J].中华实验外科杂志, 2016, 33(3):833-834. CUI Y H, WANG Y, YU Z Q.Protective effects of hydrogen sulfide against the early brain injury after subarachnoid hemorrhage in rat[J].Chinese Journal of Experimental Surgery, 2016, 33(3):833-834.(in Chinese)
[16] 曹旭, 黄若兰, 陈小武.硫化氢通过TLR4/NF-κB通路改善MPP⁺所致SH-SY5Y细胞炎症性损伤[J].华中科技大学学报(医学版), 2020, 49(4):399-403. CAO X, HUAGN R L, CHEN X W.H₂S alleviates MPP⁺-induced inflammation by inhibiting TKR4/NF-κB pathway[J].Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2020, 49(4):399-403.(in Chinese)
[17] SIDHAPURIWALA J, NG S W, BHATIA M.Effects of hydrogen sulfide on inflammation in caerulein-induced acute pancreatitis[J].Journal of Inflammation, 2009, 6(1):35.
[18] ZHANG P, LI F, WIEGMAN C H, et al.Inhibitory effect of hydrogen sulfide on ozone-induced airway inflammation, oxidative stress, and bronchial hyperresponsiveness[J].American Journal of Respiratory Cell and Molecular Biology, 2015, 52(1):129-137.
[19] WANG X, WANG Y, KONG M, et al.miR-22-3p suppresses sepsis-induced acute kidney injury by targeting PTEN[J].Bioscience Reports, 2020, 40(6):BSR20200527.
[20] 艾厚喜, 黄文婷, 王文, 等.莫诺苷抑制过氧化氢诱导SH-SY5Y神经细胞氧化损伤[J].中国康复理论与实践, 2007, 13(11):1023-1025. AI H X, HUANG W T, WANG W, et al.Morroniside inhibits H₂O₂-induced oxidative injury in SH-SY5Y neuroblastoma cells[J].Chinese Journal of Rehabilitation Theory and Practice, 2007, 13(11):1023-1025.(in Chinese)
[21] 卢春凤, 杨玉, 商宇, 等.ROS介导的氧化应激在INH诱导的细胞毒性中的作用及槲皮素的干预[J].生态毒理学报, 2015, 10(3):209-215. LU C F, YANG Y, SHANG Y, et al.The role of ROS-mediated oxidative stress in INH-induced cytotoxicity and the intervention of quercetin[J].Asian Journal of Ecotoxicology, 2015, 10(3):209-215.(in Chinese)
[22] ALLEN R G, TRESINI M.Oxidative stress and gene regulation[J].Free Radical Biology and Medicine, 2000, 28:463-499.
[23] PRASADA R, MUKTESH G, SAMANTA J, et al.Natural history and profile of selective cytokines in patients of acute pancreatitis with acute kidney injury[J].Cytokine, 2020, 133:155177.
[24] VURAL P, DEGIRMENCIOGLU S, SARAL N Y, et al.Tumor necrosis factor α (-308), interleukin-6(-174) and interleukin-10(-1082) gene polymorphisms in polycystic ovary syndrome[J].European Journal of Obstetrics & Gynecology & Reproductive Biology, 2010, 150(1):61-65.
[25] ZHU Y, XU D L, DENG F, et al.Angiotensin (1-7) attenuates sepsis-induced acute kidney injury by regulating the NF-κB pathway[J].Frontiers in Pharmacology, 2021, 12:601909.
[26] CROFFORD L J J.COX-1 and COX-2 tissue expression:Implications and predictions[J].Journal of Rhematology Supplement, 1997, 49(49):15-19.
[27] 曲道炜, 王昕冉, 冯博, 等.桂芍知母汤对佐剂性关节炎大鼠抗炎作用的机制研究[J].中药药理与临床, 2020, 36(3):6. QU D W, WANG X R, FENG B, et al.Mechanism on anti-inflammatory effect of Guishaozhimu decoction in adjuvant rats[J].Pharmacology and Clinics of Chinese Materia Medica, 2020, 36(3):6.(in Chinese)
[28] VUOLTEENAHO K, MOILANEN T, HMLINEN M, et al.Regulation of nitric oxide production in osteoarthritic and rheumatoid cartilage role of endogenous IL-1 inhibitors[J].Acta Rheumatologica Scandinavica, 2003, 32(1):19-24.
[29] LEVY Y, BROUET J.Interleukin-10 prevents spontaneous death of germinal center B cells by induction of the bcl-2 protein[J].Journal of Clinical Investigation, 1994, 93(1):424-428.
[30] CENCI E, ROMANI L, MENCACCI A, et al.Interleukin-4 and interleukin-10 inhibit nitric oxide-dependent macrophage killing of Candida albicans[J].European Journal of Immunology, 2010, 23(5):1034-1038.
[31] LI T, ZHAO B, WANG C, et al.Regulatory effects of hydrogen sulfide on IL-6, IL-8 and IL-10 levels in the plasma and pulmonary tissue of rats with acute lung injury[J].Experimental Biology and Medicine, 2008, 233(9):1081-1087.
[32] 杨永华, 王伟, 胡斌, 等.硫化氢对大鼠肢体再灌注损伤后炎症因子和线粒体能量代谢障碍的影响[J].中国医学科学院学报, 2019, 41(2):234-241. YANG Y H, WANG W, HU B, et al.Effect of hydrogen sulfide on inflammatory factors and mitochondrial energy metabolic disorders after reperfusion injury in rats[J].Acta Academiae Medicinae Sinicae, 2019, 41(2):234-241.(in Chinese)