發(fā)布時(shí)間：2018-04-19 16:46

  本文選題：膿毒癥 + 肺臟炎性損傷�。� 參考：《安徽醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：研究rhTNFR:Fc(益賽普)對(duì)膿毒癥大鼠血清中腫瘤壞死因子(TNF-α)、白細(xì)胞介素-1(IL-1)、細(xì)胞間粘附分子1(ICAM-1)、肺臟組織中核因子κB(NF-κB)的表達(dá)及肺臟組織炎癥變化的影響,探討rhTNFR:Fc(益賽普)對(duì)膿毒癥大鼠的肺臟是否具有保護(hù)作用。 方法：清潔級(jí)Sprague Dawley (SD)大鼠108只,雌雄不拘,隨機(jī)分為3組：(1)對(duì)照組(A組,n=36),膿毒癥組(B組,n=36),治療組(C組,n=36)；(2)A組36只動(dòng)物于麻醉后開(kāi)腹翻動(dòng)盲腸后關(guān)腹,在術(shù)后3h,9h.24h.36h,48h和72h六個(gè)時(shí)相點(diǎn)從腹主動(dòng)脈抽取血標(biāo)本,采用酶聯(lián)免疫分析法檢測(cè)腫瘤壞死因子(TNF-α)、白細(xì)胞介素-1(IL-1)、細(xì)胞間粘附分子-1(ICAM-1)水平；打開(kāi)胸腔,采取綠豆大小左下肺臟組織,做蛋白質(zhì)印跡實(shí)驗(yàn)(Western blot)檢測(cè)肺臟NF-kB蛋白含量,取少許左上肺組織行病理切片觀察肺臟組織炎癥變化情況。每個(gè)時(shí)相點(diǎn)6只大鼠。(3)B組動(dòng)物麻醉后行盲腸結(jié)扎穿孔(CLP)術(shù),制造膿毒癥大鼠模型,分別于術(shù)后3h,9h,24h,36h,48h和72h六個(gè)時(shí)相點(diǎn)行與對(duì)照組相同的操作步驟。(4)C組動(dòng)物行盲腸結(jié)扎穿孔(CLP)術(shù),術(shù)后立即經(jīng)尾靜脈注射rhTNFR:Fc(益賽普)2mL/kg(lg/L),于術(shù)后3h,9h,24h,36h,48h和72h六個(gè)時(shí)相點(diǎn)行與膿毒癥組相同的操作步驟。(5)所有數(shù)據(jù)以“均數(shù)±標(biāo)準(zhǔn)差”表示,采用SPSS13.0統(tǒng)計(jì)軟件包進(jìn)行分析,相關(guān)性分析采用Spearman等級(jí)相關(guān)分析。 結(jié)果： 1.血清TNF-a水平變化：膿毒癥組、治療組血清TNF-a水平在術(shù)后3h時(shí)相點(diǎn)為最大值,隨著時(shí)間推移,兩組濃度水平呈下降趨勢(shì),但仍高于假手術(shù)對(duì)照組,在CLP術(shù)后各時(shí)相點(diǎn),膿毒癥組及治療組與對(duì)照組比較均具有顯著差異性(P0.01)。治療組血清TNF-a水平較膿毒癥組低,在術(shù)后各時(shí)相點(diǎn)與膿毒癥組比較差異有顯著性(P0.05)。 2.血清IL-1水平變化：膿毒癥組、治療組血清IL-1水平在術(shù)后3h時(shí)相點(diǎn)為最大值,隨著時(shí)間推移,兩組濃度水平呈下降趨勢(shì),但仍高于假手術(shù)對(duì)照組,在CLP術(shù)后各時(shí)相點(diǎn),膿毒癥組及治療組與對(duì)照組比較均具有顯著差異性(P0.01)。治療組血清IL-1水平較膿毒癥組低,在術(shù)后各時(shí)相點(diǎn)與膿毒癥組比較差異有顯著性(P0.05)。 3.血清ICAM-1水平變化：膿毒癥組、治療組血清ICAM-1水平在術(shù)后3h時(shí)相點(diǎn)為最大值,隨著時(shí)間推移,兩組濃度水平呈下降趨勢(shì),但仍高于假手術(shù)對(duì)照組,在CLP術(shù)后各時(shí)相點(diǎn),膿毒癥組及治療組與對(duì)照組比較均具有顯著差異性(P0.01)。治療組血清ICAM-1水平較膿毒癥組低,在術(shù)后各時(shí)相點(diǎn)與膿毒癥組比較差異有顯著性(P0.05)。 4.病理切片結(jié)果顯示：假手術(shù)對(duì)照組的肺泡組織完整性好,周?chē)鸁o(wú)明顯炎性細(xì)胞浸潤(rùn)及組織水腫；膿毒癥組肺臟組織見(jiàn)肺泡組織完整性破壞,周?chē)梢?jiàn)明顯組織水腫和大量炎性細(xì)胞浸潤(rùn)；而治療組相對(duì)膿毒癥組肺泡組織結(jié)構(gòu)完整性較好,周?chē)M織水腫及炎性細(xì)胞浸潤(rùn)相對(duì)較輕。但與對(duì)照組相比有所增加。 5.Western-blot結(jié)果顯示：膿毒癥組的電泳條帶的面積明顯大于治療組和對(duì)照組,顏色也較治療組和對(duì)照組偏深。治療組的電泳條帶的面積和顏色深度在3h、72h時(shí)相點(diǎn)低于對(duì)照組,其余時(shí)相點(diǎn)均高于對(duì)照組,但較膿毒癥組低。 6. TNF-α、IL-1及ICAM-1之間的相關(guān)性分析結(jié)果： 1).TNF-α與ICAM-1具有顯著正相關(guān),相關(guān)系數(shù)為0.415, P0.05; 2). ICAM-1與IL-1具有顯著正相關(guān),相關(guān)系數(shù)為0.983, P0.01; 3). TNF-α與IL-1具有顯著正相關(guān),相關(guān)系數(shù)為0.342, P0.05。 結(jié)論： 1.在膿毒癥大鼠模型中,肺部組織NF-κB蛋白的表達(dá)量增加以及病理切片結(jié)果均提示膿毒癥大鼠肺臟組織中存在炎癥反應(yīng)過(guò)程。 2.在膿毒癥大鼠實(shí)驗(yàn)?zāi)Ｐ椭?膿毒癥大鼠血清中TNF-α、IL-1及ICAM-1等炎癥因子異常升高,是引起全身炎癥反應(yīng)和膿毒癥引起肺部炎性損傷的主要因子。 3. rhTNFR:Fc(益賽普)可以減少膿毒癥大鼠血清中TNF-α、IL-1及ICAM-1等炎癥因子的釋放,也降低了核因子κB (NF-κB)在肺臟組織中的表達(dá),可能是干擾了由膿毒癥引起的肺部炎癥反應(yīng)的核心環(huán)節(jié),從而減輕肺臟的炎性損傷。 4. rhTNFR:Fc(益賽普)作為T(mén)NF-α受體阻斷劑,可能部分阻斷了早期炎癥反應(yīng)的過(guò)程,降低了膿毒癥的炎癥反應(yīng)程度,從而對(duì)膿毒癥大鼠肺臟的炎性損傷具有保護(hù)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.
Result錛，

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