發(fā)布時間：2018-05-04 18:23

  本文選題：S100A9鈣結(jié)合蛋白 + S100A9mRNA��； 參考：《瀘州醫(yī)學(xué)院》2013年碩士論文

【摘要】：目的：觀察小鼠銅綠假單胞菌肺炎肺組織中鈣結(jié)合蛋白S100A9mRNA表達(dá)，并同時測定對應(yīng)小鼠血液中S100A9鈣結(jié)合蛋白含量，觀察肺組織的病理改變，探討鈣結(jié)合蛋白S100A9在銅綠假單胞菌所致小鼠肺部感染中的地位與作用。方法：小鼠36只，按隨機數(shù)字表分為空白對照組12只、模型組小鼠24只。通過經(jīng)環(huán)莢膜穿刺注射銅綠假單胞菌菌液建立肺部感染模型�？瞻讓φ战M（經(jīng)環(huán)莢膜注射等量的2個麥?zhǔn)蠁挝粷舛龋?×10~8cfu/ml）0.6ml的銅綠假單胞菌菌液，造模過程中死亡2只）、模型組（經(jīng)環(huán)莢膜注射等量的2個麥?zhǔn)蠁挝粷舛鹊你~綠假單胞菌菌液0.6ml，造模中死亡5只,分別在在3小時和9小時2個時間點各處死9和10只）。取小鼠右肺下葉HE染色觀察肺組織病理，觀察空白對照組，3h模型組及9h模型組的病理改變；使用普通PCR檢測系統(tǒng)對三組小鼠肺組織中S100A9mRNAPCR反應(yīng)進(jìn)程中的產(chǎn)量進(jìn)行實時監(jiān)測,最終對實驗數(shù)據(jù)進(jìn)行分析,從而比較三組小鼠S100A9mRNA表達(dá)，并同步采用雙抗體夾心酶聯(lián)免疫吸附測定(ELISA)法分別測定三組小鼠血液中S100A9以及TNF-a的含量，比較三組小鼠血液中生成S100A9鈣結(jié)合蛋白的差異及與TNF-a變化的相關(guān)系。結(jié)果：（1）病理觀察：大鼠右下肺肺組織HE染色200倍光鏡下病理結(jié)果顯示，正常對照組（1組）肺泡結(jié)構(gòu)較為均勻與清晰，有少許淋巴細(xì)胞與巨噬細(xì)胞侵潤；模型組（3h組）主要為肺泡炎性改變，肺泡內(nèi)及周圍可見滲出，中性粒細(xì)胞和淋巴細(xì)胞，巨噬細(xì)胞浸潤;模型組（9h組）肺泡炎性最嚴(yán)重，肺泡內(nèi)及周圍可見大量滲出，大量中性粒細(xì)胞和淋巴細(xì)胞，巨噬細(xì)胞浸潤，病理炎性改變明顯重于3h組。（2）普通PCR實驗:本實驗根據(jù)電泳后條帶亮度的強弱來判斷模板拷貝數(shù)的高低，或者是表達(dá)量的高低。實驗結(jié)果得出s100A9mRNA在第3h，9h組中的條帶亮度強度明顯高于空白組，提示S100A9表達(dá)量模型組明顯高于對照組,且經(jīng)統(tǒng)計分析后3h與對照組差異具有統(tǒng)計學(xué)意義（p0.05)，與9h模型組比較（P0.05）差異有意義。（3）S100A9鈣結(jié)合蛋白ELISA檢測：結(jié)果顯示3h，9h組外周血中S100A9的含量明顯高于空白組，且以3h組增高最明顯，3h組與空白組比較差異具有統(tǒng)計學(xué)意義（p0.05）與9h組比較（P0.05）有差異。（4）腫瘤壞死因子TNF-a的ELISA檢測結(jié)果顯示：3h，9h組外周血中TNF-a的含量明顯高于空白組，差異具有統(tǒng)計學(xué)意義（p0.05）。（5）通過對S100A9鈣結(jié)合蛋白與腫瘤壞死因子-a的相關(guān)性分析后，得出兩者之間的變化趨勢存在正相關(guān)，3h模型組r=0.77；9h模型組，，r=0.653.結(jié)論：（1）.鈣結(jié)合蛋白S100A9參與小鼠銅綠假單胞菌肺部炎癥反應(yīng)。（2）.鈣結(jié)合蛋白S100A9在小鼠銅綠假單胞菌肺炎中與經(jīng)典炎性因子TNF-a存在密切相關(guān)。
[Abstract]:Objective: to observe the expression of calcium-binding protein (S100A9mRNA) in the lung tissues of mice with Pseudomonas aeruginosa pneumonia, and to detect the content of S100A9 calcium binding protein in the blood of corresponding mice, and to observe the pathological changes of lung tissue. To investigate the role of calcium binding protein (S100A9) in pulmonary infection induced by Pseudomonas aeruginosa (Pseudomonas aeruginosa) in mice. Methods: 36 mice were randomly divided into control group (n = 12) and model group (n = 24). Pulmonary infection model was established by injection of Pseudomonas aeruginosa through circular capsule. Blank control group (injected with the same amount of 2 McClone unit concentrations of 6 脳 10~8cfu/ml)0.6ml Pseudomonas aeruginosa solution through circular capsule), In the model group, two rats died in the process of modeling, and in the model group, 9 and 10 rats were killed at 3 hours and 9 hours, respectively, and 5 rats were killed in the model group (injection of equal amount of 0.6 ml of Pseudomonas aeruginosa per unit concentration of 2 McClomonas aeruginosa) through the capsule of the model, and 9 hours, respectively. The histopathology of lung tissue was observed by HE staining in the lower lobe of the right lung of mice, and the pathological changes were observed in the 3 h model group and 9 h model group of the blank control group, and the output of the S100A9mRNAPCR reaction process in the lung tissues of the three groups was monitored by ordinary PCR detection system in real time. Finally, the experimental data were analyzed to compare the expression of S100A9mRNA in the three groups of mice, and the contents of S100A9 and TNF-a in blood of the three groups were measured by double antibody sandwich enzyme-linked immunosorbent assay (Elisa). To compare the difference of S100A9 calcium binding protein in blood of three groups and its relationship with the change of TNF-a. Results (1) pathological observation: the pathological results showed that the alveolar structure of the right lower lung tissue of the rats was uniform and clear, with a few lymphocytes and macrophages infiltrating into the lung tissue of the right lower lung tissue by HE staining for 200 times under the light microscope. The results showed that the alveolar structure was more uniform and clear in the normal control group (1 group). In the model group (3 h), the alveolar inflammatory changes were mainly observed, the infiltration of neutrophils, lymphocytes and macrophages was observed in and around the alveoli, and the alveolar inflammation was the most serious in the model group (9 h), and a large amount of exudation was observed in and around the alveoli. A large number of neutrophils and lymphocytes, macrophages infiltration, pathological inflammatory changes were significantly more serious than the 3h group. (2) ordinary PCR experiment: this experiment according to the intensity of the band brightness after electrophoresis to judge the template copy number, or the high or low level of expression. The results showed that the luminance intensity of s100A9mRNA was significantly higher in the 3h / 9h group than in the blank group, suggesting that the expression of S100A9 in the model group was significantly higher than that in the control group. After statistical analysis, there was a significant difference between the control group and the control group at 3h, and the difference was significant compared with the model group at 9h (P0.05). The results showed that the content of S100A9 in the peripheral blood of the 3h / 9h group was significantly higher than that of the blank group. The results of ELISA detection of tumor necrosis factor (TNF-a) showed that the level of TNF-a in peripheral blood of the 3h group was significantly higher than that of the control group (P 0.05), and that of the control group was significantly higher than that of the control group (P < 0.05). By analyzing the correlation between S100A9 calcium binding protein and tumor necrosis factor-a, it was found that there was a positive correlation between the changes of S100A9 calcium binding protein and tumor necrosis factor-a in 3h model group. Conclusion: 1: 1. Calcium binding protein (S100A9) is involved in pulmonary inflammation of Pseudomonas aeruginosa in mice. Calcium binding protein S100A9 is closely related to the classical inflammatory factor TNF-a in mouse pseudomonas aeruginosa pneumonia.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R563.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 張青,毛寶齡,錢桂生,李琦,陳正堂,徐劍鋮;油酸-內(nèi)毒素序貫致傷大鼠血漿和肺泡灌洗液TNF-α、IL-1β和IL-6水平的變化[J];第三軍醫(yī)大學(xué)學(xué)報;2004年15期

2 費敏;王選錠;;銅綠假單胞菌肺炎的最新進(jìn)展[J];國際呼吸雜志;2006年04期

3 陳霞;李建生;;S100A9蛋白的生物學(xué)特性與臨床意義[J];國外醫(yī)學(xué)(內(nèi)科學(xué)分冊);2006年06期

4 呂志敢;郭政;;腫瘤壞死因子的研究進(jìn)展[J];山西醫(yī)科大學(xué)學(xué)報;2006年03期

5 高桂新;沈華浩;;感冒雙解合劑對流感病毒FM1感染小鼠肺部炎性損傷的影響[J];中國病理生理雜志;2007年06期

本文編號：1844091