rhTNFR:FC对脓毒症大鼠肺脏的保护作用

发布时间：2018-04-19 16:46

  本文选题：脓毒症 + 肺脏炎性损伤　； 参考：《安徽医科大学》2013年硕士论文

【摘要】：目的：研究rhTNFR:Fc(益赛普)对脓毒症大鼠血清中肿瘤坏死因子(TNF-α)、白细胞介素-1(IL-1)、细胞间粘附分子1(ICAM-1)、肺脏组织中核因子κB(NF-κB)的表达及肺脏组织炎症变化的影响,探讨rhTNFR:Fc(益赛普)对脓毒症大鼠的肺脏是否具有保护作用。 方法：清洁级Sprague Dawley (SD)大鼠108只,雌雄不拘,随机分为3组：(1)对照组(A组,n=36),脓毒症组(B组,n=36),治疗组(C组,n=36)；(2)A组36只动物于麻醉后开腹翻动盲肠后关腹,在术后3h,9h.24h.36h,48h和72h六个时相点从腹主动脉抽取血标本,采用酶联免疫分析法检测肿瘤坏死因子(TNF-α)、白细胞介素-1(IL-1)、细胞间粘附分子-1(ICAM-1)水平；打开胸腔,采取绿豆大小左下肺脏组织,做蛋白质印迹实验(Western blot)检测肺脏NF-kB蛋白含量,取少许左上肺组织行病理切片观察肺脏组织炎症变化情况。每个时相点6只大鼠。(3)B组动物麻醉后行盲肠结扎穿孔(CLP)术,制造脓毒症大鼠模型,分别于术后3h,9h,24h,36h,48h和72h六个时相点行与对照组相同的操作步骤。(4)C组动物行盲肠结扎穿孔(CLP)术,术后立即经尾静脉注射rhTNFR:Fc(益赛普)2mL/kg(lg/L),于术后3h,9h,24h,36h,48h和72h六个时相点行与脓毒症组相同的操作步骤。(5)所有数据以“均数±标准差”表示,采用SPSS13.0统计软件包进行分析,相关性分析采用Spearman等级相关分析。 结果： 1.血清TNF-a水平变化：脓毒症组、治疗组血清TNF-a水平在术后3h时相点为最大值,随着时间推移,两组浓度水平呈下降趋势,但仍高于假手术对照组,在CLP术后各时相点,脓毒症组及治疗组与对照组比较均具有显著差异性(P0.01)。治疗组血清TNF-a水平较脓毒症组低,在术后各时相点与脓毒症组比较差异有显著性(P0.05)。 2.血清IL-1水平变化：脓毒症组、治疗组血清IL-1水平在术后3h时相点为最大值,随着时间推移,两组浓度水平呈下降趋势,但仍高于假手术对照组,在CLP术后各时相点,脓毒症组及治疗组与对照组比较均具有显著差异性(P0.01)。治疗组血清IL-1水平较脓毒症组低,在术后各时相点与脓毒症组比较差异有显著性(P0.05)。 3.血清ICAM-1水平变化：脓毒症组、治疗组血清ICAM-1水平在术后3h时相点为最大值,随着时间推移,两组浓度水平呈下降趋势,但仍高于假手术对照组,在CLP术后各时相点,脓毒症组及治疗组与对照组比较均具有显著差异性(P0.01)。治疗组血清ICAM-1水平较脓毒症组低,在术后各时相点与脓毒症组比较差异有显著性(P0.05)。 4.病理切片结果显示：假手术对照组的肺泡组织完整性好,周围无明显炎性细胞浸润及组织水肿；脓毒症组肺脏组织见肺泡组织完整性破坏,周围可见明显组织水肿和大量炎性细胞浸润；而治疗组相对脓毒症组肺泡组织结构完整性较好,周围组织水肿及炎性细胞浸润相对较轻。但与对照组相比有所增加。 5.Western-blot结果显示：脓毒症组的电泳条带的面积明显大于治疗组和对照组,颜色也较治疗组和对照组偏深。治疗组的电泳条带的面积和颜色深度在3h、72h时相点低于对照组,其余时相点均高于对照组,但较脓毒症组低。 6. TNF-α、IL-1及ICAM-1之间的相关性分析结果： 1).TNF-α与ICAM-1具有显著正相关,相关系数为0.415, P0.05; 2). ICAM-1与IL-1具有显著正相关,相关系数为0.983, P0.01; 3). TNF-α与IL-1具有显著正相关,相关系数为0.342, P0.05。 结论： 1.在脓毒症大鼠模型中,肺部组织NF-κB蛋白的表达量增加以及病理切片结果均提示脓毒症大鼠肺脏组织中存在炎症反应过程。 2.在脓毒症大鼠实验模型中,脓毒症大鼠血清中TNF-α、IL-1及ICAM-1等炎症因子异常升高,是引起全身炎症反应和脓毒症引起肺部炎性损伤的主要因子。 3. rhTNFR:Fc(益赛普)可以减少脓毒症大鼠血清中TNF-α、IL-1及ICAM-1等炎症因子的释放,也降低了核因子κB (NF-κB)在肺脏组织中的表达,可能是干扰了由脓毒症引起的肺部炎症反应的核心环节,从而减轻肺脏的炎性损伤。 4. rhTNFR:Fc(益赛普)作为TNF-α受体阻断剂,可能部分阻断了早期炎症反应的过程,降低了脓毒症的炎症反应程度,从而对脓毒症大鼠肺脏的炎性损伤具有保护作用。
[Abstract]:Objective: To investigate the effect of rhTNFR:Fc (rhTNFR:Fc) on the expression of tumor necrosis factor (TNF- a), interleukin -1 (IL-1), intercellular adhesion molecule 1 (ICAM-1), nuclear factor kappa B (NF- kappa B) in the lung tissues and the changes of lung tissue inflammation in the rats of sepsis, and to explore whether the rhTNFR:Fc (leonopup) has protective effects on the lungs of sepsis rats. Use.
Methods: 108 Sprague Dawley (SD) rats were randomly divided into 3 groups: (1) the control group (group A, n=36), sepsis group (B group, n=36), the treatment group (group C, n=36), and (2) the 36 animals in the A group opened the caecum after anaesthesia, and the blood samples were extracted from the abdominal aorta and used enzymes for 3h, 9h.24h.36h, and six phase points after the operation. The level of tumor necrosis factor (TNF- alpha), interleukin -1 (IL-1) and intercellular adhesion molecule -1 (ICAM-1) were detected by combined immunoassay; the thoracic cavity was opened and the lower left lung tissue of mung bean was taken. The protein content of lung NF-kB was detected by the Western blot test (Western blot), and a little left upper lung tissue was taken to observe the inflammatory changes in the lung tissue. 6 rats at each time. (3) group B animals were treated with cecal ligation and perforation (CLP), and the rat model of sepsis was made. The same operation steps were performed at the six time points of 3H, 9h, 24h, 36h, 48h and 72h, respectively. (4) the operation of the blind intestinal perforation (CLP) was performed in the group of C, and the tail vein was injected rhTNFR:Fc after the operation. 2mL/kg (lg/L), 3h, 9h, 24h, 36h, 48h and 72h were the same operation steps as the sepsis group after the operation. (5) all the data were expressed as "mean standard deviation", the analysis was carried out by the SPSS13.0 statistical package, and the correlation analysis was analyzed by the Spearman grade correlation analysis.
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