本文選題：重組人腫瘤壞死因子受體融合蛋白 + 膿毒癥�。� 參考：《安徽醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：本研究采用盲腸穿孔結(jié)扎法致大鼠膿毒癥動(dòng)物模型，觀察膿毒癥大鼠心肌損害情況，探討應(yīng)用人腫瘤壞死因子受體融合蛋白（rhTNFR：Fc）早期干預(yù)對(duì)膿毒癥心肌損害的保護(hù)作用，并初步研究其作用機(jī)理。 方法：清潔級(jí)SD大鼠108只，全為雌性，隨機(jī)分為3組：（1）假手術(shù)組（A組，n=36），膿毒癥組（B組,n=36），膿毒癥加rhTNFR：Fc組(C組,n=36)；（2）A組動(dòng)物于稱重后，麻醉，后行盲腸探查術(shù)后關(guān)閉腹腔，分別于術(shù)后3h,9h,24h,36h，48h和72h六時(shí)相點(diǎn)行腹主動(dòng)脈采血，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）、心肌肌鈣蛋白I（cTnI）水平,留取心臟組織，采用蛋白質(zhì)印跡實(shí)驗(yàn)（Western blot法）作NF-κB P65蛋白檢測(cè)，在每一時(shí)相點(diǎn)取6只大鼠（3）B組動(dòng)物稱重后，麻醉，后行盲腸結(jié)扎穿孔術(shù)后關(guān)閉腹腔，分別于術(shù)后3h,9h,24h,36h，48h和72h六時(shí)相點(diǎn)行與假手術(shù)組相同的操作步驟，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）、心肌肌鈣蛋白I（cTnI）水平,留取心臟組織，作核轉(zhuǎn)錄因子-κB（NF-κB）P65蛋白檢測(cè)，在每一時(shí)相點(diǎn)取6只大鼠；（4）C組動(dòng)物行盲腸結(jié)扎穿孔（CLP）術(shù)后，立即從頸外靜脈注入益賽普（rhTNFR：Fc）水溶液2mL／kg(1g／L)，分別于術(shù)后3h,9h,24h,36h，48h和72h六時(shí)相點(diǎn)行與假手術(shù)組相同的操作步驟，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）、心肌肌鈣蛋白I（cTnI）水平,留取心臟組織，作NF-κB P65蛋白檢測(cè)，在每一時(shí)相點(diǎn)取6只大鼠。(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-α的變化 膿毒癥加rhTNFR：Fc組與膿毒癥組血清TNF-α水平在CLP術(shù)后均較假手術(shù)組均增高（p0.05），膿毒癥加rhTNFR：Fc組在3h達(dá)峰值（257.619±5.558pg/ml）,隨后兩組均有下降趨勢(shì)，（p0.05），從3h至72h膿毒癥加rhTNFR：Fc組均明顯低于膿毒癥組（p0.05）。 2.血清IL-1的變化 與假手術(shù)組相比，膿毒癥加rhTNFR：Fc組與膿毒癥組血清IL-1水平在CLP術(shù)后均增高（p0.05），膿毒癥加rhTNFR：Fc組在3h達(dá)峰值（179.23±2.506pg/ml）,隨后兩組均有下降趨勢(shì)，（p0.05），，從3h至72h膿毒癥加rhTNFR：Fc組均明顯低于膿毒癥組（p0.05）。 3.血清cTnI的變化 與假手術(shù)組相比，膿毒癥加rhTNFR：Fc組與膿毒癥組血清cTnI水平在CLP術(shù)后均增高（p0.05），膿毒癥加rhTNFR：Fc組在3h達(dá)峰值（228.332±3.635pg/ml）,隨后兩組均有下降趨勢(shì)（p0.05），從3h至72h均明顯低于膿毒癥組（p0.05）。 4.（NF-κB）P65蛋白水平表達(dá)情況 膿毒癥加rhTNFR：Fc組與膿毒癥組CLP術(shù)后3h （NF-κB）P65蛋白水平均高于假手術(shù)組（p0.05），隨后均有下降趨勢(shì)，膿毒癥加rhTNFR：Fc組下降幅度較膿毒癥組更快,在24h，36h，48h膿毒癥加rhTNFR：Fc組（NF-κB）P65蛋白水平均低于膿毒癥組，至72h兩組無明顯差異。 5.cTnI與TNF-α、IL-1之間的相關(guān)性分析 TNF-α與cTnI具有顯著正相關(guān),相關(guān)系數(shù)為0.393，P=0.164；IL-1與cTnI具有顯著正相關(guān),相關(guān)系數(shù)為0.980，P=0.000。 結(jié)論： 1.在膿毒癥致急性心肌損傷的過程中,膿毒癥大鼠血清cTnI早期升高，反映了膿毒癥有心肌損傷，通過檢測(cè)血清cTnI水平，可以反映急性心肌損傷的嚴(yán)重程度。 2.膿毒癥發(fā)生發(fā)展過程中,膿毒癥大鼠血清TNF-α、IL-1升高，與cTnI具有高度相關(guān)性，表明炎癥因子過度表達(dá)參與了膿毒癥心肌損傷，（NF-κB）P65蛋白表達(dá)增強(qiáng)，推測(cè)膿毒癥可能通過NF-κB信號(hào)轉(zhuǎn)導(dǎo)通路致細(xì)胞因子過度表達(dá)，從而參與膿毒癥的心肌損傷的過程。 3.膿毒癥大鼠早期應(yīng)用rhTNFR：Fc干預(yù)后，cTnI下降，心肌損傷減輕，TNF-α、IL-1炎癥因子水平下降，（NF-κB）P65蛋白表達(dá)下調(diào)，說明rhTNFR：Fc通過抑制TNF-α介導(dǎo)NF-κB信號(hào)轉(zhuǎn)導(dǎo)阻斷細(xì)胞因子過度表達(dá)，調(diào)節(jié)失控性炎癥反應(yīng)，對(duì)膿毒癥心肌損傷起保護(hù)作用。
[Abstract]:Objective: in this study, the rat model of sepsis caused by cecal perforation and ligation was used to observe the myocardial damage in septic rats and to explore the protective effect of the early intervention of human tumor necrosis factor receptor fusion protein (rhTNFR:Fc) on the myocardial damage of sepsis, and the mechanism of its action was preliminarily studied.
Methods: 108 clean SD rats, all female, were randomly divided into 3 groups: (1) sham operation group (group A, n=36), sepsis group (group B, n=36), sepsis plus rhTNFR:Fc group (group C, n=36), and (2) group A, after weighing, anesthesia, and after cecum exploration, the abdominal aorta was closed after the operation of 3H, 9h, 24h, six, and six. The level of tumor necrosis factor (TNF- a), interleukin -1 (IL-1), cardiac troponin I (cTnI), cardiac tissue and NF- kappa B P65 protein were detected by enzyme linked immunosorbent assay (Western blot), and 6 rats (3), B group were weighed at each moment, anesthesia, and post cecal ligation and perforation. Closed abdominal cavity, the same operation steps as 3h, 9h, 24h, 36h, 48h and 72h six after the operation were the same as that of the sham operation group. The level of tumor necrosis factor (TNF- alpha), interleukin -1 (IL-1) and cardiac troponin I (cTnI) were detected by enzyme immunoassay, and the nuclear factor kappa B (nuclear factor kappa) protein was detected at each phase point. 6 rats were taken, and (4) after the C group was treated with cecal ligation and perforation (CLP), the Yasip (rhTNFR:Fc) aqueous solution 2mL / kg (1g / L) was injected immediately from the external jugular vein. The same operation steps as 3h, 9h, 24h, 36h, 48h and six were performed respectively after the operation, and the tumor necrosis factor (alpha) and leukocyte were detected by enzyme immunoassay. -1 (IL-1), cardiac troponin I (cTnI) level, cardiac tissue and NF- kappa B P65 protein were detected. 6 rats were taken at each phase point. (5) all data were expressed as "mean standard deviation", and SPSS13.0 statistical package was used for analysis. Correlation analysis used Spearman grade correlation analysis.
Result錛

本文編號(hào)：1833586