硫氧还蛋白-1 在急性坏死性胰腺炎大鼠中的表达及褪黑素对其的影响
Expression of pancreatic thioredoxin-1 in rats with acute necrotizing pancreatitis and effect of melatonin on thioredoxin-1
摘要目的 观察硫氧还蛋白-1(TRX-1)在急性坏死性胰腺炎(ANP)大鼠胰腺组织的表达和褪黑素干预对其的影响.方法 72只雄性SD大鼠随机分成ANP组、褪黑素组和对照组,每组24只.以腹腔注射6%L-Arginine 25 mL/kg体重3次、每次间隔1 h的方法制备ANP模型.褪黑素组在制模前30min腹腔注射0.25%褪黑素20 ml/kg体重;ANP组和对照组大鼠腹腔注射等容积生理盐水.术后6、12、24 h分批处死大鼠.抽血测定血清淀粉酶含量;取胰腺组织行病理学检查,并评分;检测胰腺组织丙二醛(MDA)、髓过氧化酶(MPO)含量;免疫组化检测胰腺组织TRX-1蛋白表达;RT-PCR检测胰腺组织TRX-1 mRNA的表达.结果 ANP组12 h点血清淀粉酶、胰腺组织MDA和MPO以及TRX-1 mRNA和蛋白表达水平分别为(3 012±1 425)u/L、(4.13±1.85)nmol/mg prot、(7.45±1.26)nmol/mg prot、0.68±0.18、66.8±8.1;褪黑素组分别为(1 835±499)U/L、(3.03±2.12)nmol/mg prot、(5.32±1.06)nmml//mgprot、0.50±0.09、80.29±8.14,两组比较均有显著差异(P<0.05).褪黑素组胰腺TRX-1 mRNA和蛋白表达峰值从ANP组的制模后12 h提前到制模后6 h.结论 ANP大鼠胰腺组织TRX-1 mRNA和蛋白表达增高;褪黑素干预能促进胰腺组织早期表达TRX-1 mRNA和蛋白,减轻胰腺组织的损伤.
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abstractsObjective To investigate the expression of pancreatic thioredoxin-1 (TRX-1) in rats with acute necrotizing pancreatitis (ANP) and the effect of pretreatment of melatonin on its expression. Methods Male Spraque-Dawley rats (n = 12) were randomly divided to ANP group, melatonin group, control group with 24 rats in each group. The rats in ANP group received three intraperitoneal injections of 25 ml/kg body weight 6% L-arginine at an interval of 1 h to induce ANP. The rats in melatonin group received intraperitoneal injections of 25 ml/kg body weight 6% melatonin 30 min before ANP induction; rats in ANP group and control group received intraperitoneal injections of same amount of saline. Rats were sacrificed at 6 h, 12 h and 24 h after ANP induction. The serum level of amylase was measured and the pathological evaluation of pancreatic tissues was performed. The concentrations of malondialdehyde (MDA) and myeloperoxidase (MPO) in pancreatic tissues were measured. The expressions of TRX-1 protein were detected by immunohistochemistry and the expressions of TRX-1 mRNA in pancreatic tissues were determined by RT-PCR.Results In ANP group, serum level of amylase, MDA, MPO, TRX-1 mRNA and TRX-1 protein in pancreatic tissues were (3 012 ±1 425) U/L, (4.13 ± 1. 85)nmol/mg prot,(7.45 ± 1.26)nmol/mg prot, 0.68 ±0. 18, 66.8 ±8. 1, while they were (1 835±499)U/L, (3.03 ±2.12) nmol/mg prot, (5. 32 ± 1.06) nmol/mg prot, 0.50±0.09, 80. 29 ±8. 14, respectively in melatonin group, the values in melatonin group were significantly lower thanthose in ANP group (P < 0.05). The peak value of TRX-1 mRNA and TRX-1 protwein expressions shifted from 12 h after ANP induction in ANP group to 6 h after ANP induction in melatonin group. Conclusions The expression of pancreatic TRX-1 protein and TRX-1 mRNA in rats with ANP was significantly increased. Melatonin pretreatment could promote pancreatic tissues to express TRX-1 protein and TRX-1 mRNA, and may be protective for pancreatic tissues damages.
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