本文關(guān)鍵詞： 急性肺損傷 腫瘤壞死因子 白細(xì)胞介素 烏司他丁 脂多糖　出處：《大連醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過(guò)觀察烏司他丁(ulinastatin，UTI)對(duì)急性肺損傷大鼠肺組織腫瘤壞死因子(TNF-α)、白細(xì)胞介素（IL-1β)、白介素1受體抗體（IL-1ra）mRNA表達(dá)的影響，探討烏司他丁治療急性肺損傷機(jī)制。 方法：將72只雄性SD大鼠隨機(jī)分為4組，即A組實(shí)驗(yàn)對(duì)照組，，B組內(nèi)毒素組和C組烏司他丁低劑量組(UTI50U/kg），D組烏司他丁高劑量組(UTI100U/kg)，制模前20分鐘給予C組和D組大鼠，分別經(jīng)腹腔注射不同劑量的烏司他丁，BCD組均采用尾靜脈注射內(nèi)毒素磷酸脂多糖（15mg/kg）制作急性肺損傷模型，A組則經(jīng)腹腔及尾靜脈注射等量的生理鹽水，各組于注射內(nèi)毒素磷酸脂多糖（LPS）后2小時(shí)、6小時(shí)、24小時(shí)各取樣6只；以1%戊巴比妥鈉50mg/kg腹腔注射麻醉，大鼠仰臥位固定于操作臺(tái)，開(kāi)胸取肺，10%甲醛固定右下肺葉組織，待行病理檢查。右下肺組織凍存于-80℃，應(yīng)用HE染色，光鏡下觀察各組大鼠肺組織變化情況，應(yīng)用引物設(shè)計(jì)軟件primer primer5.0設(shè)計(jì)引物，應(yīng)用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)（ReverseTranscription-Polymerase Chain Reaction，RTPCR）法擴(kuò)增并測(cè)定大鼠肺組織細(xì)胞TNF-α mRNA、IL-1β mRNA、IL-1ra mRNA表達(dá)情況。檢測(cè)不同劑量的烏司他丁干預(yù)下，內(nèi)毒素（LPS）致急性肺損傷大鼠肺組織TNF-α、IL-1β、IL-1ra的mRNA表達(dá)情況差異。 結(jié)果:大鼠右下肺組織經(jīng)HE染色檢查，光鏡下觀察顯示對(duì)照組大鼠肺泡結(jié)構(gòu)清晰完整，無(wú)破壞，間隔無(wú)增寬；內(nèi)毒素脂多糖(LPS)組肺組織炎癥表現(xiàn)明顯，肺泡結(jié)構(gòu)不同程度受損，肺組織間隔增厚變寬，炎細(xì)胞浸潤(rùn)明顯；烏司他丁(ulinastatin)組則顯示較內(nèi)毒素組呈現(xiàn)一定的炎癥抑制，表現(xiàn)為肺泡組織間隔變窄，炎細(xì)胞浸潤(rùn)減輕；對(duì)肺損傷大鼠肺組織內(nèi)TNF-α mRNA、IL-1β mRNA、IL-1ra mRNA表達(dá)情況，應(yīng)用RT-PCR法、凝膠電泳計(jì)算機(jī)灰度分析得出結(jié)果，經(jīng)SPSS19軟件統(tǒng)計(jì)分析，應(yīng)用方差分析及t檢驗(yàn)，分別比較同一時(shí)間點(diǎn)不同處理組的TNF-α表達(dá)情況的組間差異，結(jié)果提示烏司他丁干預(yù)組與內(nèi)毒素組之間統(tǒng)計(jì)學(xué)差異顯著(P0.001)，烏司他丁低劑量組與高劑量組相比較，差異也顯著(P0.001)，再分別比較IL-1β mRNA、IL-1ra mRNA表達(dá)情況，應(yīng)用烏司他丁組與內(nèi)毒素組比較均有顯著差異(P0.005)，烏司他丁低劑量組與高劑量組相比較，差異也顯著(P0.001)。 結(jié)論:烏司他丁對(duì)于膿毒癥引起的急性肺損傷具有一定的治療作用，其機(jī)制可能為降低以TNF-α IL-1β IL-1ra為代表的炎癥因子表達(dá)，抑制肺部炎癥反應(yīng)的發(fā)生。
[Abstract]:Objective: To observe the effect of ulinastatin (Ulinastatin, UTI) on the expression of tumor necrosis factor (TNF- alpha), interleukin (IL-1 beta) and interleukin 1 receptor antibody (IL-1ra) mRNA in lung tissue of rats with acute lung injury, and to explore the mechanism of ulinastatin in the treatment of acute lung injury.
Methods: 72 male SD rats were randomly divided into 4 groups: A group, experimental control group, B group, LPS group and ulinastatin group C low dose group (UTI50U/kg), group D Ulinastatin high dose group (UTI100U/kg), mold 20 minutes before giving C group and D group rats respectively, by intraperitoneal injection of different doses of ulinastatin, group BCD by intravenous injection of endotoxin lipopolysaccharide (15mg/kg) phosphate production model of acute lung injury in A group with normal saline intraperitoneal and intravenous injection of the same amount of phosphate groups in the injection of endotoxin lipopolysaccharide (LPS) after 2 hours, 6 hours, 24 hours the sampling of 6; with 1% 50mg/kg intraperitoneal injection of pentobarbital sodium anesthesia, rats supine fixed on the operation table, lung, 10% formalin fixed to the lower lobe of the right lung tissue, lung tissue for pathological examination. The right under the freezing at -80 deg.c, using HE staining, observe the changes of lung tissue of rats under the light microscope, using primer Design software primer Primer5.0 primers by reverse transcriptase polymerase chain reaction (ReverseTranscription-Polymerase Chain Reaction, RTPCR) were amplified by TNF- and determination of mRNA cells in lung tissue of rats with IL-1 alpha, beta mRNA, IL-1ra mRNA expression. The effects of different doses of statin intervention detection, endotoxin (LPS) injury of TNF- alpha, lung tissue of rats with acute the expression of IL-1ra induced lung IL-1 beta, mRNA difference.
Results: the lung tissue of rats after right HE staining, light microscope observation showed that rat alveolar structure of control group was clear and complete, no damage, no interval widened; lipopolysaccharide (LPS) group showed obvious inflammation of lung tissue, alveolar structure damaged to varying degrees, lung septum thick and wide, infiltration of inflammatory cells obviously; ulinastatin (Ulinastatin) group showed endotoxin group showed a suppression of inflammation, tissue showed alveolar septum, infiltration of inflammatory cells to reduce; lung injury in rats TNF- mRNA IL-1 alpha, beta mRNA, IL-1ra mRNA expression, using RT-PCR method, gel electrophoresis and computer gray analysis results by SPSS19 software, statistical analysis, analysis of variance and t test, respectively, compared with the same time point of different groups of TNF- expression differences between the groups. The results suggest that the effects between the intervention group and endotoxin group he d statistics Significant differences (P0.001), ulinastatin low dose group and high dose group compared with significant difference (P0.001), and then were compared with IL-1 IL-1ra beta mRNA, mRNA expression and application of ulinastatin group and endotoxin group were significantly different (P0.005), ulinastatin low dose group and high dose compared to the group, a significant difference (P0.001).
Conclusion: Ulinastatin has certain therapeutic effect on acute lung injury caused by sepsis, and its mechanism may be to reduce the expression of inflammatory factors represented by TNF- alpha IL-1 beta IL-1ra and inhibit the occurrence of pulmonary inflammatory reaction.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R563.8

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