多聚腺苷酸二磷酸核糖聚合酶抑制剂对重症急性胰腺炎肾损伤的作用

Effect of poly adenosine diphasphate ribose polymerase inhibitor on renal injury in rats with severe acute pancreatitis

  • 摘要: 目的 探讨多聚腺苷酸二磷酸核糖聚合酶(PARP)抑制剂3氨基苯甲酰胺(3AB)对重症急性胰腺炎(SAP)肾损伤的作用。方法 雄性Wistar大鼠56只,分为7组:假手术组(简称SO组),SAP(3、6、12 h)组,3AB+SAP(3、6、12 h)组,每组8只。采用5%牛磺胆酸钠逆行胰胆管注射法制备SAP模型,3AB+SAP组在SAP造模前30 min经股静脉注射3AB(20 μg/g)。检测血清淀粉酶、肾功能、肾髓过氧化物酶(MPO)水平,光镜观察胰腺和肾脏病理变化,Western blot检测肾组织PARP活化程度(PAR蛋白表达水平代表PARP活性程度)、细胞内黏附分子1 (ICAM1)和P选择素蛋白表达。多个样本均数比较采用单因素方差分析,两组间比较采用t检验。肾损伤分级采用KruskalWallis法秩和检验。结果 SAP(3、6、12 h)组血清淀粉酶分别为(3806±229)U/L、(4898±295)U/L和(5726±372)U/L,高于3AB+SAP相应时点组的(2785±160)U/L、(3241±198)U/L和(3953±249)U/L,差异有统计学意义(t=3.652,4.672,4.407,P<0.05);SAP(3、6、12 h)组尿素氮分别为(11.6±0.8)mmol/L、(19.3±1.3)mmol/L和(29.6±2.1)mmol/L,均高于3AB+SAP相应时点组的(7.5±0.5)mmol/L、(10.5±0.7)mmol/L和(21.6±1.5)mmol/L,其中6、12 h时相点两组比较,差异有统计学意义(t=3.836,6.849,P<0.05);SAP(3、6、12 h)组肌酐分别为(48.7±3.1)μmol/L、(58.3±3.7)μmol/L和(75.9±5.4)μmol/L,均高于3AB+SAP相应时点组的(40.7±2.6)μmol/L、(43.2±2.6)μmol/L和(53.4±3.2)μmol/L,其中6、12 h时相点两组比较,差异有统计学意义(t=3.279,3.073,P<0.05);SAP(3、6、12 h)组肾脏MPO值分别为(0.69±0.06)U/g、(1.07±0.09)U/g和(1.42±0.13)U/g,均高于3AB+SAP相应时点组的(0.57±0.05)U/g、(0.75±0.06)U/g和(0.89±0.07)U/g,其中6、12 h时相点两组比较,差异有统计学意义(t=3.066,4.012,P<0.05)。SAP(3、6、12 h)组胰腺病理评分分别为(6.50±0.53)分、(9.06±0.66)分和(11.75±0.89)分, 显著高于3AB+SAP相应时点组的(4.25±0.31)分、(6.06±0.51)分和(7.57±0.59)分,差异有统计学意义(t=3.631,3.598,5.147,P<0.05)。SO组肾脏结构正常,SAP(12 h)组可见肾小球淤血性改变,细胞界限模糊,肾小管上皮细胞坏死脱落,间质出血、管腔变窄或闭塞,炎症细胞浸润,而3AB+SAP(12 h)组上述病理改变均减轻,病理分级降低(P<0.05)。SO组肾组织PAR、ICAM1和P选择素蛋白的相对表达量分别为1.00±0.21、1.00±0.18和1.00±0.16,显著低于SAP(12 h)组的3.83±0.63、5.42±0.83和3.71±0.48,差异有统计学意义(t=6.955,23.107,10.352,P<0.05),而3AB+SAP(12 h)组肾组织PAR、ICAM1和P选择素蛋白相对表达量分别为1.94±0.36、2.35±0.35和2.11±0.29,较SAP(12 h)组显著下降,差异有统计学意义(t=3.977,12.115,5.012,P<0.05)。结论 PARP抑制剂3AB对SAP肾损伤有保护作用。其作用可能与抑制肾组织ICAM1和P选择素蛋白表达以及减缓中性粒细胞浸润程度有关。

     

    Abstract: Objective To investigate the effects and mechanisms of poly adenosine diphosphate (ADP) ribose polymerase inhibitor 3aminobenzamide (3AB) on kidney injury in rates with severe acute pancreatitis (SAP). Methods Fiftysix male Wistar rats were divided into the sham operation (SO) group, SAP (3, 6, 12 hours) groups, and 3AB+SAP (3, 6, 12 hours) groups, and there were 8 rats in each group. SAP model was established by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. Rats in the 3AB+SAP group were infused with 3AB (20 μg/g) via femoral vein 30 minutes before SAP model establishment. The serum amylase, kidney function and renal myeloperoxidase (MPO) were determined, and pathological scores of pancreatic and renal tissues were evaluated under light microscope. Renal poly ADPribose formation, intercellular adhesion molecules1 (ICAM1) and Pselectin expression were detected by the Western blot. All data were analyzed using the analysis of variance or t test. Renal injury grading was analyzed using the KruskalWallis nonparametric test. Results The levels of serum amylase of SAP 3, 6, 12 groups were (3806±229)U/L, (4898±295)U/L and (5726±372)U/L, which were significantly higher than (2785±160)U/L, (3241±198)U/L and (3953±249)U/L of the 3AB+SAP groups (t=3.652, 4.672, 4.407, P<0.05). The levels of blood urea nitrogen were (11.6±0.8)mmol/L, (19.3±1.3)mmol/L and (29.6±2.1)mmol/L, which were higher than (7.5±0.5)mmol/L, (10.5±0.7)mmol/L and (21.6±1.5)mmol/L of the 3AB+SAP groups. There were significant differences in the levels of blood urea nitrogen between the SAP group and the 3AB+SAP group at the 6 and 12 hours (t=3.836, 6.849, P<0.05). The levels of creatinine of the SAP 3, 6, 12 hours groups were (48.7±3.1)μmol/L, (58.3±3.7)μmol/L and (75.9±5.4)μmol/L, which were higher than (40.7±2.6)μmol/L, (43.2±2.6)μmol/L and (53.4±3.2)μmol/L of the 3AB+SAP groups. There were significant differences in the levels of creatinine between the SAP group and the 3AB+SAP group at the 6 and 12 hours (t=3.279, 3.073, P<0.05). The renal MPO activity of the SAP 3, 6, 12 hours groups were (0.69±0.06)U/g, (1.07±0.09)U/g and (1.42±0.13)U/g, which were higher than (0.57±0.05)U/g, (0.75±0.06)U/g and (0.89±0.07)U/g of the 3AB+SAP groups. There were significant differences in the renal MPO activity between the SAP group and the 3AB+SAP group at the 6 and 12 hours (t=3.066, 4.012, P<0.05). The pancreatic pathological scores of the SAP 3, 6 and 12 hours group were 6.50±0.53, 9.06±0.66 and 11.75±0.89, which were significantly higher than 4.25±0.31, 6.06±0.51 and 7.57±0.59 of the 3AB+SAP group (t=3.631, 3.598, 5.147, P<0.05). The structure of the kidney was normal in the SO group. Congestive changes were observed in glomerulus of kidney, the renal tubular epithelial cell was necrosed, and luminal narrowing or occlusion, hemorrhage in the intercellular substance and inflammatory cell infiltration were observed in the SAP 12 hours group. The pathological changes of the 3AB+SAP 12 hours group were significantly slighter (P<0.05). The relative expressions of poly ADPribose, ICAM1 and Pselectin of the SO group were 1.00±0.21, 1.00±0.18, 1.00±0.16, which were significantly lower than 3.83±0.63, 5.42±0.83, 3.71±0.48 of the SAP 12 hours group (t=6.955, 23.107, 10.352, P<0.05). The relative expressions of polyADPribose, ICAM1 and Pselectin of the 3AB+SAP 12 hours group were 1.94±0.36, 2.35±0.35, 2.11±0.29, which were significantly lower than SAP 12 hours group (t=3.977, 12.115, 5.012, P<0.05). Conclusions Poly ADPribose polymerase inhibitor 3AB protects kidney from injury in the experimental SAP rats. Poly ADPribose polymerase inhibitor 3AB functions by suppressing the ICAM1 and Pselectin expression and reducing neutrophil infiltration.

     

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