本文選題：脂多糖 + 急性肺損傷��； 參考：《中南大學(xué)》2013年碩士論文

【摘要】：目的：通過(guò)檢測(cè)脂多糖誘導(dǎo)的急性肺損傷(ALI)大鼠肺組織中高遷移率蛋白1(HMGB1) mRNA及其蛋白表達(dá)水平,以及甲基強(qiáng)的松龍注射劑干預(yù)后對(duì)急性肺損傷大鼠肺組織HMGB1表達(dá)的影響,探討HMGB1在脂多糖誘導(dǎo)的急性肺損傷中的作用,以及甲基強(qiáng)的松龍注射劑干預(yù)后在急性肺損傷中的表達(dá)變化。 方法：清潔級(jí)雄性SD大鼠54只,隨機(jī)分成3組,每組18只；(1)正常對(duì)照組(N組)：每只尾靜脈注射生理鹽水2ml/kg,1小時(shí)后再尾靜脈注射生理鹽水1ml/kg;(2)急性肺損傷組(ALI組)：尾靜脈注射脂多糖6mg/kg,1小時(shí)后再尾靜脈注射生理鹽水1ml/kg;(3)甲基強(qiáng)的松龍干預(yù)組(MPN組)：每只尾靜脈注射脂多糖6mg/kg,1小時(shí)后再尾靜脈注射甲基強(qiáng)的松龍20mg/kg。觀察各組大鼠行為改變,三組分別于注射后12h、24h、36h采集標(biāo)本,每個(gè)時(shí)相點(diǎn)6只大鼠。采集動(dòng)脈血?dú)?放血后取部分肺組織行HE染色,觀察肺組織病理學(xué)改變,并測(cè)定干濕比重(W/D)；采用免疫組織化學(xué)法和Real-Time PCR法檢測(cè)大鼠肺組織HMGB1表達(dá)水平。 結(jié)果：(1)各組一般狀況的變化：ALI組大鼠12h后出現(xiàn)毛發(fā)豎直,蜷作一團(tuán),呼吸急促,爪趾發(fā)紺,反應(yīng)遲鈍,萎靡厭食,排便減少或性狀改變,24h最明顯,36h癥狀有所減輕；MPN組癥狀輕于同時(shí)點(diǎn)ALI組。 (2)各組干濕比重的變化：ALI組大鼠肺干濕比重(W/D)明顯增加,各時(shí)相點(diǎn)與N組比較差異均有顯著統(tǒng)計(jì)學(xué)意義(P0.01),但各時(shí)相點(diǎn)之間無(wú)統(tǒng)計(jì)學(xué)差異(P0.05), MPN組肺干濕比重(W/D)增高,與N組比較有統(tǒng)計(jì)學(xué)意義(P0.01), MPN組與ALI組比較肺干濕比重(W/D)下降。 (3)各組血?dú)庾兓篘組P02正常,ALI組大鼠血?dú)夥治鯬02明顯下降(P0.05), MPN組PO2下降,同一時(shí)相點(diǎn)較ALI組減輕。 (4)各組肺組織病理改變：N組大鼠肺組織小葉結(jié)構(gòu)清晰,肺泡腔及肺泡間質(zhì)無(wú)炎性細(xì)胞浸潤(rùn)。ALI組肺組織呈不同程度腫脹,表面散在紫紅色淤血斑,肺泡壁間隔增寬,肺泡及肺血管明顯充血水腫,肺泡腔內(nèi)見(jiàn)紅細(xì)胞及炎性細(xì)胞滲出,肺間質(zhì)也見(jiàn)大量炎癥細(xì)胞浸潤(rùn),且隨著時(shí)間的延長(zhǎng),肺組織病理改變逐漸加重,24h最明顯,36h有所減輕。MPN組較同一時(shí)相點(diǎn)ALI組肺組織改變有不同程度的減輕。 (5)各組肺組織中HMGB1表達(dá)變化：正常組中大鼠肺組織有少量HMGB1mRNA及其蛋白表達(dá),ALI組和MPN組大鼠肺組織中HMGB1mRNA及其蛋白表達(dá)水平明顯高于N組(均p0.05),且隨著時(shí)間延長(zhǎng)呈逐漸增加趨勢(shì),24h HMGB1mRNA表達(dá)達(dá)峰值；同一時(shí)相點(diǎn)ALI組大鼠肺組織HMGB1mRNA及其蛋白表達(dá)水平高于MPN組(P0.05)。 結(jié)論： 1、急性肺損傷中肺組織HMGB1mRNA及其蛋白表達(dá)明顯升高,且24h時(shí)達(dá)高峰,36h下降但仍高于N組,提示HMGB1在炎癥后期持續(xù)性高水平表達(dá),在ALI發(fā)生、發(fā)展過(guò)程中尤其是ALI后期失控性炎癥反應(yīng)過(guò)程中可能發(fā)揮重要作用。 2、甲基強(qiáng)的松龍注射劑干預(yù)后急性肺損傷中肺組織：HMGB1mRNA及其蛋白表達(dá)較急性肺損傷組有所降低,可有效改善大鼠急性肺損傷的炎癥反應(yīng),對(duì)脂多糖所致的急性肺損傷有一定的保護(hù)作用。
[Abstract]:Objective: to detect the expression of high mobility protein 1 (HMGB1) mRNA and protein in lung tissue of rats with acute lung injury (ALI) induced by lipopolysaccharide, and the effect of methylprednisolone injection on the expression of HMGB1 in lung tissue of rats with acute lung injury, and explore the role of HMGB1 in the acute lung injury induced by lipopolysaccharide, as well as the effect of HMGB1 on the acute lung injury induced by lipopolysaccharide. The expression of basal prednisolone injection in acute lung injury.
Methods: 54 clean male SD rats were randomly divided into 3 groups, 18 rats in each group, and (1) normal control group (group N): each caudal vein was injected with saline 2ml/kg, and 1ml/kg was injected into the tail vein after 1 hours; (2) the acute lung injury group (ALI group): the tail vein was injected with lipopolysaccharide 6mg/kg, and the tail vein was injected with saline 1ml/kg after 1 hours; (3) a The MPN group (group MPN): each tail vein was injected with lipopolysaccharide 6mg/kg, and after 1 hours the tail vein was injected with methylprednisolone 20mg/kg. to observe the behavior changes in each group. The three groups were collected respectively after the injection of 12h, 24h, 36h and 6 rats at each time phase. The blood gas was collected and the lung tissue was stained with HE after blood letting and the lung was observed. The changes of histopathology and dry wet weight (W/D) were measured. The expression level of HMGB1 in lung tissue was detected by immunohistochemistry and Real-Time PCR.
Results: (1) the changes in the general condition of each group: the rats in group ALI had hair vertical, curled up, breathing quickly, cyanosis of claw toes, slow reaction, weary anorexia, reduced defecation or change of character, 24h was most obvious, 36h symptoms were lightened, and MPN group was less than group ALI in MPN.
(2) the change of dry and wet specific gravity of each group: the lung dry and wet specific gravity (W/D) of the ALI group increased significantly, and the difference of the phase points with the N group had significant statistical significance (P0.01), but there was no statistical difference between the phase points (P0.05), and the proportion of lung and humidity (W/D) in the MPN group increased, and was statistically significant (P0.01), and the lung dry and wet ratio was compared with the ALI group. The MPN and ALI groups compared the lung dry wet ratio. The weight (W/D) drops.
(3) blood gas changes in each group: P02 in group N was normal, blood gas analysis P02 in group ALI decreased significantly (P0.05), PO2 in MPN group decreased, and the same time point decreased compared with ALI group.
(4) pathological changes of lung tissue in each group: the structure of pulmonary lobule in group N was clear, pulmonary alveolus and alveolar interstitial free cell infiltration in group.ALI, the lung tissue was swollen in varying degrees, the surface scattered in the purple red congestion spot, the alveolar septum widened, the alveolar and pulmonary blood vessels were obviously congested and edema, the alveolar and inflammatory cells exuded and the pulmonary interstitium was found. There was also a large number of inflammatory cells infiltrating, and with the prolongation of time, the pathological changes of lung tissue were gradually aggravated, 24h was the most obvious, and 36h was relieved in.MPN group as compared with the same phase point ALI group.
(5) the change of HMGB1 expression in lung tissue of each group: there was a small amount of HMGB1mRNA and protein expression in the lung tissue of the normal group. The expression level of HMGB1mRNA and protein in lung tissue of group ALI and MPN rats was significantly higher than that of group N (P0.05), and the expression of 24h HMGB1mRNA expression was gradually increasing with time, and the same phase point ALI group rats were in the same time. The expression level of HMGB1mRNA and protein in lung tissue was higher than that in group MPN (P0.05).
Conclusion:
1, the expression of HMGB1mRNA and its protein in lung tissue in acute lung injury increased significantly, and reached the peak at 24h, but 36h decreased but still higher than that in group N, suggesting that HMGB1 may be expressed at a high level in the late stage of inflammation, which may play an important role in the process of ALI, especially in the process of uncontrolled inflammatory reaction in the late stage of ALI.
2, after the methylprednisolone injection, the expression of HMGB1mRNA and its protein expression in acute lung injury is lower than that in the acute lung injury group. It can effectively improve the inflammatory response of acute lung injury in rats and protect the acute lung injury caused by lipopolysaccharide.
【學(xué)位授予單位】：中南大學(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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本文編號(hào)：1983998