髓樣細(xì)胞觸發(fā)受體-1（TREM-1）在大腸埃希菌誘導(dǎo)新生大鼠敗血癥中的意義
發(fā)布時間：2018-12-11 07:04
【摘要】：目的:敗血癥是臨床常見重癥之一,為感染后所致的全身炎癥反應(yīng)綜合癥。新生兒對各種病原微生物易感性高,極易引起敗血癥,且病死率很高,敗血癥的早期診斷、及時合理治療是目前臨床所需迫切面對的問題。固有免疫是一種天然免疫系統(tǒng),是機(jī)體抵御病原體感染的第一道防線。巨噬細(xì)胞是機(jī)體固有免疫系統(tǒng)的重要一員,其功能狀態(tài)直接影響到感染的預(yù)后。髓樣細(xì)胞觸發(fā)受體-1(Triggering receptor expressed on myeloid cells-1,TREM-1)選擇性表達(dá)于中性粒細(xì)胞、單核巨噬細(xì)胞表面,能夠促進(jìn)TNF-α、IL-1β等細(xì)胞因子的表達(dá),具有激發(fā)和放大炎癥反應(yīng)的作用,其可溶性的形式sTREM-1存在于體液中,共同參與炎癥反應(yīng),發(fā)揮抗炎作用。本課題通過建立新生大鼠大腸埃希菌敗血癥模型來分析組織巨噬細(xì)胞表面TREM-1的表達(dá)情況以及外周血中sTREM-1水平的變化,來探討TREM-1在新生大鼠敗血癥發(fā)病機(jī)制中的作用,并嘗試開拓?cái)⊙Y免疫調(diào)控干預(yù)的新思路。 方法:選擇無特定病原體(SPF級)7日齡Sprague-Dawley(SD)大鼠72只,體重16-20g,雌雄不限,分成對照組和敗血癥組,每組36只。所有動物按照隨機(jī)的原則分入兩組不同的時間點(diǎn),即實(shí)驗(yàn)后第2、4、8、12、24、48小時六個時間點(diǎn),每個時間點(diǎn)為6只新生大鼠。采用腹腔注射大腸埃希菌(E.coli)方法建立敗血癥模型,對照組則用等量生理鹽水腹腔注射。于各目標(biāo)時間點(diǎn)麻醉動物后,心臟采血,再留取肝臟、肺臟于福爾馬林液中固定備用。選擇免疫組織化學(xué)法分析肝臟、肺臟巨噬細(xì)胞表面TREM-1的表達(dá)情況,ELISA法檢測血漿中sTREM-1的水平。 結(jié)果:1.對照組新生大鼠肝、肺組織中巨噬細(xì)胞表面TREM-1和血漿中sTREM-1各時間點(diǎn)表達(dá)水平均無顯著性差異(P0.05);2.敗血癥組新生大鼠肝、肺組織中巨噬細(xì)胞表面TREM-1和血漿中sTREM-1的表達(dá)水平,均隨著實(shí)驗(yàn)時間的延長而逐漸升高,于12小時左右達(dá)高峰,后有所下降,但至48小時仍維持在較高水平;3.實(shí)驗(yàn)進(jìn)展至2小時,敗血癥組肝、肺組織中巨噬細(xì)胞表面TREM-1表達(dá)及血漿中sTREM-1濃度分別與對照組比較均無顯著性差異(P0.05);實(shí)驗(yàn)進(jìn)展至4小時及以后,敗血癥組高于對照組水平,差異有顯著統(tǒng)計(jì)學(xué)意義(P0.01)。 結(jié)論:1.隨著實(shí)驗(yàn)進(jìn)展,敗血癥組和對照組新生大鼠肝、肺組織巨噬細(xì)胞表面TREM-1表達(dá)水平和血漿中sTREM-1濃度均存在顯著性差異,說明TREM-1在敗血癥發(fā)病機(jī)制中具有重要作用。2.敗血癥組新生大鼠組織巨噬細(xì)胞表面TREM-1分子表達(dá)水平隨著敗血癥的進(jìn)展,呈時間依賴性升高,提示促炎反應(yīng)加劇;而血漿中sTREM-1濃度亦升高,提示機(jī)體抗炎反應(yīng)亦增強(qiáng)。3.巨噬細(xì)胞表面TREM-1和血漿中sTREM-1水平呈平行同向變化,是機(jī)體促炎反應(yīng)和抗炎反應(yīng)力爭達(dá)到動態(tài)平衡的重要標(biāo)志之一。4.據(jù)此,TREM-1可作為敗血癥診斷的一種新標(biāo)志物,并嘗試開拓?cái)⊙Y免疫調(diào)控干預(yù)的新思路。
[Abstract]:Objective: septicemia is one of the common clinical severe cases, which is caused by systemic inflammatory response syndrome. Newborns are susceptible to various pathogens and are prone to septicemia, and the mortality is very high. The early diagnosis and timely and reasonable treatment of septicemia are urgent problems that need to be faced in clinical practice. Innate immunity is a innate immune system and the first line of defense against pathogen infection. Macrophages are important members of the innate immune system, and their functional status directly affects the prognosis of infection. Myeloid cell trigger receptor-1 (Triggering receptor expressed on myeloid cells-1,TREM-1) is selectively expressed on the surface of neutrophils and mononuclear macrophages, which can promote the expression of cytokines such as TNF- 偽 and IL-1 尾. The soluble form of sTREM-1 exists in the body fluid and participates in the inflammatory reaction and plays an anti-inflammatory role. In this study, we established a septicemia model of Escherichia coli in neonatal rats to analyze the expression of TREM-1 on macrophages and the changes of sTREM-1 level in peripheral blood. To explore the role of TREM-1 in the pathogenesis of septicemia in neonatal rats, and try to explore a new idea of immune regulation and intervention in septicemia. Methods: 72 Sprague-Dawley (SD) rats of 7 days age without specific pathogens (SPF grade) were divided into control group and septicemia group with 36 rats in each group. All the animals were divided into two different time points according to the principle of randomness, that is, the 2nd hour, 812h, 24h, 48h after the experiment, each time point was 6 newborn rats. The septicemia model was established by intraperitoneal injection of Escherichia coli (E.coli), while the control group was injected intraperitoneally with the same amount of normal saline. After anaesthetized animals at each target time point, blood was taken from the heart, liver was taken and lung was fixed in formalin solution. The expression of TREM-1 on the surface of hepatic and pulmonary macrophages was analyzed by immunohistochemical method and the level of sTREM-1 in plasma was detected by ELISA method. Results: 1. There was no significant difference in the expression of TREM-1 on the surface of macrophages and sTREM-1 in plasma at different time points in the control group (P0.05). In septicemia group, the expression of TREM-1 on the surface of macrophages and sTREM-1 in plasma increased gradually with the prolongation of experimental time, and reached its peak at about 12 hours, and then decreased. However, it remained at a relatively high level at 48 hours. 3. When the experiment progressed to 2 hours, there was no significant difference in the expression of TREM-1 on the surface of macrophages and the concentration of sTREM-1 in plasma between the septicemia group and the control group (P0.05). The level of septicemia was significantly higher in septicemia group than that in control group after 4 hours (P0.01). Conclusion: 1. With the progress of the experiment, there were significant differences in the expression of TREM-1 on the surface of macrophages and the concentration of sTREM-1 in plasma between the septicemia group and the control group. It is suggested that TREM-1 plays an important role in the pathogenesis of sepsis. 2. In septicemia group, the expression of TREM-1 molecules on macrophages in neonatal rats increased in a time-dependent manner with the progression of septicemia, suggesting that the pro-inflammatory response was aggravated. The concentration of sTREM-1 in plasma was also increased, indicating that the anti-inflammatory response was also increased by 3. 3%. The changes of TREM-1 on macrophage surface and sTREM-1 level in plasma are parallel, which is one of the important markers of pro-inflammatory and anti-inflammatory response to achieve dynamic balance. Therefore, TREM-1 can be used as a new marker in the diagnosis of septicemia, and try to develop a new idea of immune regulation and intervention in septicemia.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363
本文編號：2372108
[Abstract]:Objective: septicemia is one of the common clinical severe cases, which is caused by systemic inflammatory response syndrome. Newborns are susceptible to various pathogens and are prone to septicemia, and the mortality is very high. The early diagnosis and timely and reasonable treatment of septicemia are urgent problems that need to be faced in clinical practice. Innate immunity is a innate immune system and the first line of defense against pathogen infection. Macrophages are important members of the innate immune system, and their functional status directly affects the prognosis of infection. Myeloid cell trigger receptor-1 (Triggering receptor expressed on myeloid cells-1,TREM-1) is selectively expressed on the surface of neutrophils and mononuclear macrophages, which can promote the expression of cytokines such as TNF- 偽 and IL-1 尾. The soluble form of sTREM-1 exists in the body fluid and participates in the inflammatory reaction and plays an anti-inflammatory role. In this study, we established a septicemia model of Escherichia coli in neonatal rats to analyze the expression of TREM-1 on macrophages and the changes of sTREM-1 level in peripheral blood. To explore the role of TREM-1 in the pathogenesis of septicemia in neonatal rats, and try to explore a new idea of immune regulation and intervention in septicemia. Methods: 72 Sprague-Dawley (SD) rats of 7 days age without specific pathogens (SPF grade) were divided into control group and septicemia group with 36 rats in each group. All the animals were divided into two different time points according to the principle of randomness, that is, the 2nd hour, 812h, 24h, 48h after the experiment, each time point was 6 newborn rats. The septicemia model was established by intraperitoneal injection of Escherichia coli (E.coli), while the control group was injected intraperitoneally with the same amount of normal saline. After anaesthetized animals at each target time point, blood was taken from the heart, liver was taken and lung was fixed in formalin solution. The expression of TREM-1 on the surface of hepatic and pulmonary macrophages was analyzed by immunohistochemical method and the level of sTREM-1 in plasma was detected by ELISA method. Results: 1. There was no significant difference in the expression of TREM-1 on the surface of macrophages and sTREM-1 in plasma at different time points in the control group (P0.05). In septicemia group, the expression of TREM-1 on the surface of macrophages and sTREM-1 in plasma increased gradually with the prolongation of experimental time, and reached its peak at about 12 hours, and then decreased. However, it remained at a relatively high level at 48 hours. 3. When the experiment progressed to 2 hours, there was no significant difference in the expression of TREM-1 on the surface of macrophages and the concentration of sTREM-1 in plasma between the septicemia group and the control group (P0.05). The level of septicemia was significantly higher in septicemia group than that in control group after 4 hours (P0.01). Conclusion: 1. With the progress of the experiment, there were significant differences in the expression of TREM-1 on the surface of macrophages and the concentration of sTREM-1 in plasma between the septicemia group and the control group. It is suggested that TREM-1 plays an important role in the pathogenesis of sepsis. 2. In septicemia group, the expression of TREM-1 molecules on macrophages in neonatal rats increased in a time-dependent manner with the progression of septicemia, suggesting that the pro-inflammatory response was aggravated. The concentration of sTREM-1 in plasma was also increased, indicating that the anti-inflammatory response was also increased by 3. 3%. The changes of TREM-1 on macrophage surface and sTREM-1 level in plasma are parallel, which is one of the important markers of pro-inflammatory and anti-inflammatory response to achieve dynamic balance. Therefore, TREM-1 can be used as a new marker in the diagnosis of septicemia, and try to develop a new idea of immune regulation and intervention in septicemia.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363
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