硫化氢对尿源性脓毒血症肾组织细胞因子表达的影响

发布时间：2018-06-10 08:42

本文选题：硫化氢 + 尿源性脓毒血症　；参考：《南华大学》2014年硕士论文

【摘要】：目的建立尿源性脓毒血症兔模型；通过应用硫化氢干预,比较兔肾组织细胞因子的表达，明确外源性硫化氢对脓毒血症兔模型肾组织细胞因子的影响，从而揭示硫化氢在尿源性脓毒血症肾组织中发挥作用的可能机制。方法 1、家兔左侧输尿管内注入大肠杆菌（ATCC25922），并结扎左侧输尿管远端，模拟上尿路急性梗阻并感染所致脓毒血症兔模型。 2、将48只健康雄性家兔随机分成4组，每组12只。正常组（Control组）；假手术组（Sham组）；脓毒血症组（Sepsis组）；硫氢化钠组（NaHS组）。 3、每组动物按时间点又分为术后12小时、术后24小时、术后36小时、术后48小时4个时间点。分别于每个时间点观测肛温（RT）、呼吸（RR）、心率（HR）。 4、术前、术后12小时、术后24小时、术后36小时、术后48小时分别采血行外周血细胞计数分析（WBC、中性粒细胞）、肾功能（Cr、BUN）测定。 5、术后12小时、24小时、36小时、48小时各组家兔肾组织分别行HE染色，光学显微镜下观察形态结构变化。Western blot法检测各组肾组织TNF-α、IL-10的蛋白表达。RT-PCR（reverse translation-PCR）法检测NF-κB的基因表达。结果 1、一般情况：sham组的体温、心率、呼吸频率在各个时间点上与control组比较，变化不大（P0.05）；随造模时间的延长，sepsis组明显高于control、sham组（P0.05），并呈上升趋势，NaHS组略低于sepsis组（P0.05）。 2、血液检测指标：各组术前的血细胞计数（WBC、中性粒细胞）、Cr、Bun比较，差异无统计学意义。control组的血细胞计数（WBC、中性粒细胞）、Cr、Bun在12h、24h、36h、48h与sham组比较，差异无统计学意义（P0.05）；随着造模时间的延长，sepsis组明显高于control、sham组（P0.05），并呈上升趋势，NaHS组与sepsis组比较，有下降趋势，其差异有统计学意义（P0.05）。 3、病理形态学：HE染色光镜下可见sepsis组肾组织大量炎症细胞浸润，，肾小球萎缩、肾小囊扩张，肾小管管腔变大。NaHS组较sepsis组上述变化有所减轻。 4、Western Blot法检测肾组织TNF-α、IL-10的蛋白表达：Control组肾组织TNF-α的蛋白表达和sham组相比较，差异无统计学意义（P0.05）；sepsis组明显高于control、sham组，随造模时间延长，呈上升趋势（P0.05）；NaHS组与sepsis组比较，有下降趋势（P0.05）。 Control组肾组织IL-10的蛋白表达和sham组相比较，差异无统计学意义（P0.05）；sepsis组明显高于control、sham组，随造模时间延长，呈上升趋势（P0.05）；NaHS组与sepsis组比较，仍呈现上升趋势（P0.05）。 5、RT-PCR（reverse translation-PCR）法检测NF-κB的基因表达：Control组NF-κB的基因表达和sham组相比较，差异无统计学意义（P0.05）；sepsis组高于control、sham组，随造模时间延长，呈现上升趋势（P0.05）；NaHS组与sepsis组比较，有下降趋势（P0.05）。结论 1、成功建立尿源性脓毒血症兔模型。 2、外源性硫化氢抑制尿源性脓毒血症模型肾组织NF-κB及TNF-α的表达，上调IL-10来减轻尿源性脓毒血症肾损伤。
[Abstract]:Objective to establish a rabbit model of urine-derived sepsis, compare the expression of cytokines in renal tissues of rabbits with hydrogen sulfide, and clarify the effect of exogenous hydrogen sulfide on renal cytokines in rabbits with sepsis. Methods 1. Escherichia coli ATCC 25922 was injected into the left ureter of rabbits and the distal end of the left ureter was ligated. The rabbit model of sepsis caused by acute upper urinary tract obstruction and infection was simulated. 2 48 healthy male rabbits were randomly divided into 4 groups with 12 rabbits in each group. Control group in normal group; Sham group in sham group; sepsis group in sepsis group; sodium sulfide group in NaHS group. Each group was divided into 4 time points according to the time points: 12 hours after operation, 24 hours after operation, 36 hours after operation and 48 hours after operation. At each time point, we observed the anal temperature, RRV, HR4, 12 hours before operation, 24 hours after operation, 36 hours after operation, and at each time point, we observed RRV, RRV and HR4 before operation, 24 hours after operation and 36 hours after operation, respectively. Blood samples were collected 48 hours after operation for the analysis of WBC, neutrophils and renal function CrBUN.5. the renal tissues of each group were stained by HE staining after 12 hours, 24 hours, 36 hours and 48 hours after operation, respectively. The expression of TNF- 伪 IL-10 in renal tissue was detected by optical microscope. Western blot method was used to detect the expression of NF- 魏 B. results 1. The body temperature, heart rate and respiratory rate of the control group were compared with those of the control group at different time points. With the prolongation of the modeling time, the blood count in sepsis group was significantly higher than that in control sham group, and the increase trend was slightly lower in NaHS group than that in sepsis group. The blood cell count and neutrophils were compared in each group before operation. There was no significant difference in blood cell count between control group and control group. There was no significant difference between control group and sham group at 12 h, 24 h, 36 h or 48 h, but with the prolongation of modeling time, the blood cell count in control group was significantly higher than that in control sham group (P 0.05), and there was an increasing trend between NaHS group and sepsis group. There was a downward trend, and the difference was statistically significant (P 0.05). The pathological morphology showed that there were a large number of inflammatory cells infiltrating, glomerular atrophy, and renal microcapsule dilatation in sepsis group under microscope. Compared with sham group, the protein expression of TNF- 伪 IL-10 in renal tissue was detected by Western blot. There was no significant difference between control group and sham group, and the expression of TNF- 伪 protein in control group was significantly higher than that in control group, and with the prolongation of modeling time, the expression of TNF- 伪 protein in renal tissue of the control group was significantly higher than that in the control group, and the protein expression of TNF- 伪 IL-10 protein in the renal tissue of the control group was significantly higher than that of the control group. Compared with sepsis group, the expression of IL-10 protein in renal tissue of the control group was significantly higher than that of the control group compared with the control group. The expression of IL-10 protein in the control group was significantly higher than that in the controlsham group. With the prolongation of the modeling time, the expression of IL-10 protein in the renal tissue of the control group was significantly higher than that in the control group, and there was an increasing trend between the P0.05NaHS group and the sepsis group. The gene expression of NF- 魏 B was detected by RT-PCR reverse translation (PCR- PCR- PCR) method. There was no significant difference between the control group and the sham group. There was no significant difference between the control group and the control group. With the prolongation of the modeling time, the expression of NF- 魏 B gene in the control group was higher than that in the control group. With the prolongation of the modeling time, the expression of NF- 魏 B gene in the nahs group was higher than that in the sepsis group, and the expression of NF- 魏 B gene in the control group was higher than that in the control group. Conclusion 1. The rabbit model of urine-derived sepsis was successfully established. 2. Exogenous hydrogen sulfide inhibited the expression of NF- 魏 B and TNF- 伪 in renal tissue of the model of urogenic sepsis, and upregulated IL-10 to alleviate renal injury of urine-derived sepsis.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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