肿瘤坏死因子-α诱导蛋白6治疗减轻百草枯中毒大鼠急性肾损伤

Tumor necrosis factor-α induced protein 6 attenuates acute kidney injury following paraquat poisoning in rats

目的 探讨肿瘤坏死因子-α诱导蛋白6（TSG-6）对百草枯中毒大鼠急性肾损伤（AKI）的作用.方法 24只雄性SD大鼠按随机数字表法分为假手术组、模型组和TSG-6干预组,每组8只.腹腔注射等体积稀释的百草枯溶液50 mg/kg染毒制备百草枯中毒模型;假手术组则给予无菌生理盐水2 mg/kg; TSG-6干预组染毒后1h腹腔注射30 μg重组人TSG-6.各组于染毒6h后取血评估肾功能,活杀取肾组织进行病理学评估并计算AKI评分,采用实时逆转录-聚合酶链反应（RT-PCR）检测肾组织促炎症细胞因子白细胞介素（IL-1β、IL-6）和肿瘤坏死因子-α（TN F-α）的基因表达.结果 与假手术组比较,模型组血清尿素氮（BUN）、肌酐（Cr）水平及AKI评分均明显升高[BUN（mmol/L）：22.64±2.36比7.09±0.65,t=6.986,P=0.000; Cr（μmol/L）：177.28±18.67比60.32±3.11,t=7.134,P=0.000;AKI评分（分）：9.14±0.28比0.30±0.23,t=9.013,P=0.000],肾组织IL-1β、IL-6和TNF-α的mRNA表达明显升高（IL-1β mRNA：3.23±0.28比1.00±0.07,t=5.874,P=0.000; IL-6 mRNA：4.16±0.37比1.00±0.08,t=7.125,P=0.000; TNF-α mRNA：3.85±0.31比1.00±0.10,t=6.342,P=0.000）.TSG-6干预组血清BUN、Cr、AKI评分及肾组织IL-1β、IL-6、TNF-α的mRNA表达均明显低于模型组[BUN（mmol/L）：14.07±5.23比22.64±2.36,t=2.533,P=0.026; Cr（μmol/L）：112.76±14.81比177.28±18.67,t=2.778,P=0.016;AKI评分（分）：5.35±0.19比9.14±0.28,t=2.885,P=0.013;IL-1β mRNA：2.26±0.19比3.23±0.28,t=2.457,P=0.023; IL-6 mRNA：2.92±0.29比4.16±0.37,t=2.975,P=0.011;TNF-α mRNA：2.58±0.23比3.85±0.31,t=2.564,P=0.019].结论 TSG-6干预可通过抑制炎症反应减轻百草枯中毒大鼠的AKI.

Objective To explore the effects of tumor necrosis factor-α induced protein 6 （TSG-6） on acute kidney injury （AKI） following paraquat poisoning in rats.Methods Twenty-four male Sprague-Dawley （SD） rats were randomly divided into sham group （n=8）,model group （n=8） and TSG-6-treated group （n=8） using a randomized number table.Rats were given an injection of 50 mg/kg of paraquat intraperitoneally （total volume was equalled to sterile normal saline） in model and TSG-6-treated groups.Rats in sham group were given 2 mg/kg of sterile saline.Mter 1 hour of paraquat administration,rats were treated with 30 μg of recombinant human TSG-6 intraperitoneally in TSG-6-treated group.After 6 hours of paraquat administration,serum was collected to assess renal function,then rats were sacrificed and renal tissues were immediately harvested.AKI score was evaluated by renal histopathology and gene expression of pro-inflammatory cytokines including interleukins （IL-1β and IL-6） and tumor necrosis factor-α （TNF-α） in kidney was assayed with real-time reverse transcription-polymerase chain reaction （RT-PCR）.Results Compared with sham group,blood urea nitrogen （BUN）,creatinine （Cr） and AKI score were significandy increased in model group [BUN （mmoUL）：22.64 ±2.36 vs.7.09 ±0.65,t=6.986,P=0.000; Cr （μmol/L）：177.28 ± 18.67 vs.60.32 ± 3.11,t=7.134,P=0.000; AKI score：9.14 ± 0.28 vs.0.30 ± 0.23,t=9.013,P=0.000].Moreover,the mRNA expressions of IL-1β,IL-6 and TNF-α were significantly elevated in model group （IL-1β mRNA：3.23 ± 0.28 vs.1.00 ±0.07,t=5.874,P=0.000; IL-6 mRNA：4.16 ±0.37 vs.1.00 ±0.08,t=7.125,P=0.000; TNF-α mRNA：3.85 ±0.31 vs.1.00 ±0.10,t=6.342,P=0.000）.However,serum BUN,Cr,AKI score and the mRNA expressions of IL-1β,IL-6 and TNF-α in TSG-6-treated group were significantly lower than those in model group [BUN （mmol/L）：14.07 ± 5.23 vs.22.64 ± 2.36,t=2.533,P=0.026; Cr （μmol/L）：112.76 ± 14.81 vs.177.28 ± 18.67,t=2.778,P=0.016; AKI score：5.35 ±0.19 vs.9.14 ±0.28,t=2.885,P=0.013; IL-1β mRNA：2.26 ± 0.19 vs.3.23 ±0.28,t=2.457,P=0.023; IL-6 mRNA：2.92 ±0.29 vs.4.16 ±0.37,t=2.975,P=0.011; TNF-α mRNA：2.58 ± 0.23 vs.3.85 ± 0.31,t=2.564,P=0.019].Conclusion TSG-6 attenuates AKI following paraquat poisoning by suppressing inflammatory response.