PPARγ激动剂罗格列酮对大鼠肾脏再灌注损伤的保护作用

Protective effect of PPAR-γ agonist rosiglitazone on renal ischemia-reperfusion injury

目的探讨罗格列酮(Rosiglitazone,RGZ)对大鼠肾脏缺血再灌注损伤的保护作用及作用机制。方法将30只大鼠按照双盲法随机均分成假手术组(Sham)、缺血性肾损伤组(IRI)和罗格列酮(RGZ)组,各10只。利用血管夹夹闭左侧肾蒂,去除右肾诱导大鼠肾缺血再灌注损伤模型,缺血45 min,再灌注4 h。ELISA检测血清血清肌酐(Cr)、尿素氮(BUN)、肿瘤坏死因子(TNF-α)、白介素-8(IL-8)和白介素-6(IL-6)表达水平;RT-PCR检测TNF-α、IL-8和IL-6 mRNA水平;硫代巴比妥酸(TBA)法检测丙二醛(MDA)的含量;Western blot检测PPAR-γ和pPPAR-γ表达水平;黄嘌呤氧化酶法检测过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)活性;过碘酸雪夫氏(PAS)染色检测肾组织病理形态。结果 IRI组血清Cr和BUN分别为(160.15±25.17)μmol/L和(90.22±16.17)mmol/L,较RGZ组的(47.16±5.13)μmol/L和(23.11±3.28)mmol/L明显增高,差异均有统计学意义(P<0.05);IRI组肾脏病理损伤评分为(3±0.5)分,较RGZ组的(1±0.5)分明显增高,差异具有统计学意义(P<0.05);IRI组血清中TNF-α、IL-8和IL-6表达水平分别为(1 400±350)pg/mL、(1 200±300)pg/mL、(1 150±250)pg/mL,较RGZ组的(650±150)pg/mL、(450±120)pg/mL和(550±170)pg/mL明显增高,差异均具有统计学意义(P<0.05);IRI组肾脏中TNF-α、IL-8和IL-6 m RNA表达水平分别为(14.00±55)、(11.00±4.3)、(9.50±2.8),较RGZ组的(5.50±1.7)、(6.40±2.1)和(3.80±1.3)明显增高,差异均具有统计学意义(P<0.05);IRI组肾脏中MDA含量为(18.12±4.15)nmol/mg,较RGZ组[(7.12±1.15)nmol/mg明显增高,差异有统计学意义(P<0.05);IRI组肾脏中CAT、GPX和SOD活性分别为(8.77±1.52)U/mg、(7.22±1.03)U/mg和(6.54±1.22)U/mg,较RGZ组的(25.17±3.22)U/mg、(23.42±3.07)U/mg和(21.22±2.79)U/mg明显增高,差异均具有统计学意义(P<0.05);IRI组肾脏中p-PPAR-γ含量为(0.32±0.07),较RGZ组的(0.51±0.11)明显降低,差异具有统计学意义(P<0.05);IRI组与RGZ组P PAR-γ表达差异无统计学意义(P>0.05)。结论 PPAR-γ激动剂罗格列酮对大鼠肾缺血再灌注损伤具有保护作用,能够减少肾脏病理改变,其作用机制与抑制氧化应激和炎症反应相关。

Objective To explore the protective effect and potential mechanism of rosiglitazone （RGZ） on renal ischemia reperfusion injury. Methods Thirty male Sprague-Dawley （SD） rats were randomly equally divided into 3 groups： Sham group, ischemia-reperfusion injury （IR1） group, and RGZ group. The left renal pedicle was clamped with vascular clamp, and the model of renal ischemia-reperfusion injury was induced by removing right kidney with 45 minutes of ischemia and 4 hours of reperfusion. The expression of creatinine （Cr）, blood urea nitrogen （BUN）, tumor necrosis factor alpha （TNF-α）, interleukin-6 （IL-6）, and interleukin-8 （IL-8） in the serum were determined by en- zyme-linked immunosorbent assay （ELISA）. The mRNA level IL 8, TNF-α, and IL-6 in the kidney were evaluated by RT-PCR. The content of malondialdehyde （MDA） was by measured by thiobarbituric acid （TBA） method. The expression of peroxisome proliferator-activated receptor T （PPAR T） and p-PPAR-T were evaluated by Western blot. The activ- ities of catalase （CAT）, glutathione peroxidase （GPx） and superoxide dismutase （SOD） were measured by xanthine oxi dase. The kidney morphology was evaluated by periodic acid-Schiff （PAS）. Results The serum expression level of Cr and BUN in the IRI group were respectively （160.15±25.17） gmol/L and （90.22±16.17） mmol/L, which were significantly higher than （47.16±5.13） μmol/L and （23.11±4.3.28） mmol/L in the RGZ group （P〈0.05）. The renal pathological damage score in the IR1 group was （3±0.5） points, which was significantly higher （1±0.5） points in the RGZ group （P〈0.05）. The serum expression level ofTNF -α, IL-8 and IL-6 in the IR1 group were （1 400±350） pg/mL, （1 200±300） pg/mL, （1 150± 250） pg/mL, respectively, which were all significantly higher than corresponding （650±150） pg/mL, （450±120） pg/mL, （550±170） pg/mL in the RGZ group （P〈0.05）. The mRNA expression levels of TNF-α, IL-8, and IL-6 in the IRI group were （14.00±5.5）, （11.00±4.3）, （9.50±2.8）, which were all significantly higher than corresponding （5.50± 1.7）, （6.40± 2.1）, （3.80±1.3） in the RGZ group （P〈0.05）. The content of MDA in the IRI group was （18.12 ± 4.15） nmol/mg, which was significantly higher than （7.12±1.15） nmol/mg in the RGZ group （P〈0.05）. The activation of CAT, GPX and SOD in the IRI group were （8.77±1.52） U/rag, （7.22±1.03） U/rag, and （6.54±1.22） U/mg, which were all significantly lower than （25.17±3.22） U/mg, （23.42±3.07） U/rag, （21.22±2.79） U/mg in the RGZ group （P〈0.05）. The content of p-PPAR y in the IRI group was （0.32±0.07）, which was lower than （0.51±0.11） in the RGZ group （P〈0.05）. There was no difference between the IRI group and the RGZ group in the expression of PPAR-T （P〉0.05）. Conclusion PPAR-3, agonist rosiglitazone can attenuate renal iscmia reperfusion injury with reducing the renal pathological changes. The mechanism of which is related to inhibition of oxidative stress and inflammatory reaction.