乌司他丁对尿源性SIRS猪的治疗作用及对肾组织NF-κB表达的影响

发布时间：2018-09-17 07:17

【摘要】：目的：观察乌司他丁注射液对尿源性SIRS猪的治疗效果并探讨乌司他丁对尿源性SIRS猪肾组织核因子（NF-κB）表达的影响。方法：健康家猪48头随机分为假手术对照组、SIRS模型组和乌司他丁治疗组3组，后两组又分为术后4h、6h、12h和24h4个时间点亚组。应用生化分析仪行肾功能测定（Cr、β2-MG、BUN）。采用Wester blot法检测肾组织中NF-κB的活性，采用SP法检测肾组织NF-κB蛋白的表达，应用RT-PCR法检测肾组织中NF-κBp65mRNA的表达，通过透射电镜观察肾组织的形态结构变化。结果： SIRS组和对照组12小时、24小时肌酐比较，有统计学意义(P0.05)，SIRS组与对照组6小时、12小时、24小时Bun、β2 MG比较，有统计学意义(P0.01)。随造模时间的延长，SIRS组动物血清Cr、Bun、β2 MG升高，Cr于术后24小时达到(122.00±11.87) μmol/L, Bun于术后24小时达到10.05±0.65mmol/L, β2 MG于术后12小时达最大值4.14±0.38mg/L。乌司他丁组血清Cr、Bun在24小时时间点，β2 MG于12小时及24小时比SIRS组降低，差异有统计学意义.对照组动物手术后Cr、Bun、β2 MG略有上升，，但和术前相比，差异无统计学意义(P0.05)。3组动物术前、0小时Cr、Bun、β2 MG比较，无统计学意义(P0.05)。RT-PCR检测肾组织中NF-κB p65mRNA的表达：对照组与各组比较，差异无统计学意义(P0.05)。乌司他丁治疗组较模型组的NF-κB p65mRNA的表达在术后12h显著降低（P0.01）；SIRS组肾组织NF-κB p65mRNA表达显著高于Sham组（P0.01），24h到达最大值。Western blot检测蛋白表达：SIRS模型组猪肾组织NF-κB蛋白的表达显著高于假手术组（P0.01），于24h到达最大值；乌司他丁治疗组较模型组的蛋白的表达在术后12h显著降低（P0.01）。免疫组织化学检测蛋白表达：模型组猪肾组织的表达显著高于假手术组（P0.01），于24h到达最大值；乌司他丁治疗组较模型组的蛋白的表达在术后12h显著降低（P0.01）。电镜观察对照组肾组织结构无明显改变；SIRS组随时间延长，肾小球足突细胞融合，肾小球内皮细胞和系膜细胞明显增生并有水肿，线粒体嵴减少，结构排列紊乱、空泡形成，有断裂或缺失，甚至崩解；UTI组肾组织的病理损害明显减轻，肾小球瘀血程度较轻。结论：乌司他丁可能通过抑制NF-κB的合成与释放，减轻肾组织病理损害,改善肾功能，而达到治疗尿源性SIRS的作用。
[Abstract]:Aim: to observe the therapeutic effect of ulinastatin injection on urogenic SIRS pigs and to investigate the effect of ulinastatin on the expression of NF- 魏 B in renal tissue of SIRS pigs. Methods: Forty-eight healthy domestic pigs were randomly divided into sham operation control group (Sirs model group) and ulinastatin treatment group (group 3). The latter two groups were divided into two groups: 4 h, 6 h, 12 h and 24h4 time point subgroup. Renal function was measured by biochemical analyzer (Cr, 尾 2-MGG bun). The activity of NF- 魏 B in renal tissue was detected by Wester blot assay, the expression of NF- 魏 B protein in renal tissue was detected by SP method, the expression of NF- 魏 Bp65mRNA in renal tissue was detected by RT-PCR method, and the morphological changes of renal tissue were observed by transmission electron microscope. Results: there was significant difference in creatinine between SIRS group and control group at 12 hours and 24 hours (P0.05). There was significant difference in Bun, 尾 2 MG between Sirs group and control group at 6 hours and 12 hours and 24 hours (P0.01). In Sirs group, the serum Cr,Bun, 尾 _ 2 MG increased and Cr reached (122.00 卤11.87) 渭 mol/L, Bun at 24 hours after operation (10.05 卤0.65 mmol / L), and 尾 _ 2 MG reached a maximum value of 4.14 卤0.38 mg / L at 12 hours after operation. The serum Cr,Bun of ulinastatin group was significantly lower than that of SIRS group at 12 hours and 24 hours, and the difference was statistically significant. The expression of Cr,Bun, 尾 2 MG in the control group was slightly increased after operation, but there was no significant difference between the control group and the preoperative group (P0.05). There was no statistical significance (P0.05). RT-PCR was used to detect the expression of NF- 魏 B p65mRNA in the renal tissue: compared with the control group, the expression of NF- 魏 B p65mRNA in the control group was significantly higher than that in the control group. The difference was not statistically significant (P0.05). The expression of NF- 魏 B p65mRNA in ulinastatin group was significantly lower than that in model group at 12 h after operation (P0.01). The expression of NF- 魏 B p65mRNA in renal tissue of SIRS group was significantly higher than that in Sham group (P0.01). The expression of NF- 魏 B protein in porcine kidney tissue of SIRS group was significantly higher than that of sham operation group (P0.01), and reached the maximum at 24 h. The expression of protein in ulinastatin group was significantly lower than that in model group at 12 h after operation (P0.01). The expression of protein in the model group was significantly higher than that in the sham-operated group (P0.01), and reached the maximum at 24 hours, and the protein expression in the ulinastatin group was significantly lower than that in the model group at 12 h after operation (P0.01). Electron microscope observation showed that there was no obvious change in renal tissue structure in Sirs group. In Sirs group, glomerular podocyte cells fused, glomerular endothelial cells and Mesangial cells proliferated and edema, mitochondrial cristae decreased, structure was disordered and vacuoles were formed. In UTI group, the pathological damage of renal tissue was alleviated, and the degree of glomerular blood stasis was mild. Conclusion: ulinastatin may inhibit the synthesis and release of NF- 魏 B, alleviate the pathological damage of renal tissue and improve renal function, so as to achieve the therapeutic effect of urine-derived SIRS.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7

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