LPS干预对军团菌肺炎作用的实验研究
发布时间：2018-11-15 17:44
【摘要】：目的 1.通过动物实验探讨LPS干预对TLR4表达的影响。 2.通过比较LPS干预后TLR4蛋白及其信号转导通路中相关细胞因子的表达,结合病理切片的结果,探讨LPS干预对军团菌感染的影响。 方法 1.以C3H/HeN小鼠(6-8周龄)作为实验动物,每组8只,雌雄各半。设立LPS干预染菌组、LPS未干预染菌组、.对照组三个实验组。首先对LPS干预组小鼠利用大肠杆菌脂多糖,以1mg/kg的剂量,进行腹腔注射干预,其他组腹腔注射生理盐水。24h后与LPS未干预组一起气管注射军团菌悬液染菌,对照组给予等量的生理盐水,分别于染菌后12h、24h、48h处死小鼠。 2.摘除眼球取血约1ml,采用密度梯度离心分离外周血单核细胞(peripheral blood mononuclear cel, PBMC),FCMS检测各组中PBMC TLR4水平,比较三组各时间点TLR4蛋白表达水平的变化；并取左肺上叶约0.02g组织固定做病理切片。 3.取左肺中叶约0.02g组织匀浆,ELISA法检测各组肺组织中TNF-a和IL-1β的含量。 结果 ①肺组织病理切片结果显示：LPS干预染菌组、LPS未干预染菌组均出现中性粒细胞、嗜酸粒细胞、单核巨噬细胞等炎性细胞,且肺泡血管有出血现象。同时两染菌组在24h可见肺组织炎性渗出液,对照组未出现炎症反应,表明对照组未受到染菌组的污染。LPS干预染菌组肺泡血管出血较少,且炎性浸润略轻于LPS未干预染菌组。经析因分析LPS干预染菌组、LPS未干预染菌组和对照组之间肺脏脏器系数差异有统计学意义(F=5.699,P=0.006),各个时间点之间差异有统计学意义(F=10.648,P0.001),分组和时间点之间交互作用差异无统计学意义(F=0.668,P0.05)。经LSD法两两比较,LPS干预染菌组和LPS未干预染菌组的脏器系数均高于对照组(P0.05),但LPS干预染菌组与LPS未干预染菌组间无统计学差异。 ②三组的TLR4蛋白水平表达有统计学差异(F=47.144,P0.001),各个时间点差异有统计学意义(F=141.543,P0.001),分组和时间点之间交互作用差异也有统计学意义(F=29.902,P0.001)。经LSD法两两比较,LPS干预染菌组TLR4‘表达水平在12h、24h与对照组相比,差异有统计学意义,且均高于对照组(P0.05)；LPS未干预组TLR4表达水平在12h高于对照组(P0.05)。LPS干预染菌组与LPS未干预染菌组TLR4蛋白均在12h达到高峰,且LPS干预组TLR4水平高于LPS未干预组(P0.05),同时两染菌组TLR4水平随着染菌时间的延长而下降,在24h、48h两染菌组TLR4表达水平差别无统计学意义。 ③三组间TNF-α浓度差异具有统计学意义(F=25.817,P0.001),时间的主效应也具有统计学意义(F=8.3933,P0.001),分组和时间点之间未发现交互作用(F=I.460,P0.05)。经两两比较,LPS干预染菌组与对照组未发现统计学差异,但LPS未干预染菌组与对照组、LPS干预染菌组之间的TNF-α浓度差异具有统计学意义,LPS未干预染菌组各个时间点TNF-α的分泌水平均高于LPS干预染菌组,两染菌组TNF-α浓度均在感染24h时达到高峰后开始下降。三组间IL-1β浓度差异具有统计学意义(F=18.990,P0.001),时间的主效应无统计学意义(F=I.323,P0.05),分组和时间点之间也未发现交互作用(F=0.382,P0.05)。LSD法两两比较后发现,LPS干预染菌组与对照组的IL-1β浓度未发现统计学差异,但LPS未干预染菌组与对照组、LPS干预染菌组之间的IL-1β浓度差异具有统计学意义,LPS未干预染菌组各个时间点IL-1β的分泌水平均高于LPS干预染菌组。 结论 ①LPS干预可显著调高小鼠PBMC TLR4表达水平,且具有短时效应。 ②LPS未干预组染菌组TNF-α、IL-1β浓度均高于对照组,提示军团菌感染后机体可能通过上调了TNF-α和IL-1β的表达水平以抵抗军团菌感染。但LPS干预组染菌组TNF-α、IL-1β浓度均与对照组无明显差别,说明LPS耐受可能会减轻军团菌感染的炎性症状。
[Abstract]:Purpose 1. The expression of TLR4 by LPS intervention was discussed by animal experiment. Influence. 2. By comparing the expression of the related cytokines in the TLR4 protein and its signal transduction pathway after LPS intervention, the results of the pathological sections were combined to study the effect of LPS on Legionella. infection Effect of method 1. C3H/ HeN mice (6-8 weeks old) as an experimental activity In each group, 8 animals and half of each group were male and female. The LPS intervention group was established. and LPS did not interfere with the stain. In the control group, three experimental groups were divided into three groups: first, the mice of the LPS intervention group were treated with E. coli lipopolysaccharide, and the injection of the abdominal cavity was carried out at a dose of 1 mg/ kg, and the other groups were injected with normal saline in the abdominal cavity. After 24 h, the suspension of the Legionella pneumophila was injected with the LPS without intervention group, and the control group was given the same amount of physiological saline, respectively. after the staining of the bacteria for 12h, The expression of TLR4 in the peripheral blood mononuclear cells (PBMC) and peripheral blood mononuclear cells (PBMC) were measured by density gradient centrifugation, and the expression levels of TLR4 protein in each group were compared in three groups. 0. 02g of tissue was fixed to pathological section. 3. The lung of each group was detected by ELISA in the middle of the left lung. organization The contents of TNF-a and IL-1 were found in the rats. Results The results of the pathological sections of the lung tissue of the rats showed that the LPS intervention group and LPS did not interfere with the bacterial group, and the neutrophils and eosinophils were found. The inflammatory cells, such as single-core macrophages, and the blood vessels of the alveoli were bleeding. At the same time, the two groups of bacteria were found in the pulmonary tissue inflammatory exudation liquid at the same time, and the control group was used as the control group. No inflammatory reaction was observed in the group, indicating that the control group was not contaminated by the stain group. The LPS interfered with the pulmonary alveolar blood of the bacterial group. There was a significant difference in lung organ coefficient between LPS and control group (F = 5.699, P = 0. 006), and there was a significant difference between the time points (F = 10.648, P0.001), group and time. point-to-point interaction There was no significant difference (F = 0. 668, P0.05). Compared with the control group (P <0.05), the organ coefficients of the LPS-treated group and the LPS-free group were higher than those in the control group (P0.05). There was no significant difference in the level of TLR4 protein in the three groups (F = 47. 144, P 0.001), and the difference of time point was statistically significant (F = 141.543, P0.001), and the interaction between the group and time point was statistically significant (F = 141.543, P 0.001). The difference was also of statistical significance (F = 29. 902, P0. The expression level of TLR4 in LPS-treated group was significantly higher than that of control group (P <0.05). The expression of TLR4 in LPS-free group was higher than that of control group (P <0.05), and the level of TLR4 expression in LPS-free group was higher than that of control group (P0.05). The TLR4 protein in both group and LPS group reached the peak at 12h, and the level of TLR4 in LPS group was higher than that of LPS group (P0.05). There was no significant difference in the expression of TLR4 in the two groups. The difference between the three groups was statistically significant (F = 25. 817, P 0.001). The primary effect of time was also of statistical significance (F = 8.3933, P 0.001). There was no interaction between the group and the time point (F = I. 460, P0.05). There was a significant difference in the concentration of TNF-1 between the group and the LPS-treated group, and the levels of TNF-1 in the LPS-free group were higher than that of the LPS-treated group. The concentration of TNF-1 in the two groups was decreased after 24 h of infection. There was no significant difference in the concentration of IL-1 in the three groups (F = 18. 990, P. 001). The primary effect of time was not significant (F = I. 323, P0.05). There was no interaction between the group and the time point (F = 0.382, P0.05). After the LSD method, the levels of IL-1 in the LPS-treated group and the control group were not found to be statistically different, but the LPS did not interfere with the control group and the control group. There was no significant difference in the concentration of IL-1 in the group and LPS intervention group, and LPS did not interfere. bacteria The levels of IL-1 at all time points in the group were higher than that of the LPS-treated group. The expression level of TLR4 in the high-dose mice was significantly increased by LPS intervention and short-time effect. The concentration of TNF-1 and IL-1 in the group-infected cell group was higher than that of the control group. After the infection of Legionella, the expression level of TNF-1 and IL-1 was up-regulated in order to resist the infection of Legionella.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.1
本文编号：2333996
[Abstract]:Purpose 1. The expression of TLR4 by LPS intervention was discussed by animal experiment. Influence. 2. By comparing the expression of the related cytokines in the TLR4 protein and its signal transduction pathway after LPS intervention, the results of the pathological sections were combined to study the effect of LPS on Legionella. infection Effect of method 1. C3H/ HeN mice (6-8 weeks old) as an experimental activity In each group, 8 animals and half of each group were male and female. The LPS intervention group was established. and LPS did not interfere with the stain. In the control group, three experimental groups were divided into three groups: first, the mice of the LPS intervention group were treated with E. coli lipopolysaccharide, and the injection of the abdominal cavity was carried out at a dose of 1 mg/ kg, and the other groups were injected with normal saline in the abdominal cavity. After 24 h, the suspension of the Legionella pneumophila was injected with the LPS without intervention group, and the control group was given the same amount of physiological saline, respectively. after the staining of the bacteria for 12h, The expression of TLR4 in the peripheral blood mononuclear cells (PBMC) and peripheral blood mononuclear cells (PBMC) were measured by density gradient centrifugation, and the expression levels of TLR4 protein in each group were compared in three groups. 0. 02g of tissue was fixed to pathological section. 3. The lung of each group was detected by ELISA in the middle of the left lung. organization The contents of TNF-a and IL-1 were found in the rats. Results The results of the pathological sections of the lung tissue of the rats showed that the LPS intervention group and LPS did not interfere with the bacterial group, and the neutrophils and eosinophils were found. The inflammatory cells, such as single-core macrophages, and the blood vessels of the alveoli were bleeding. At the same time, the two groups of bacteria were found in the pulmonary tissue inflammatory exudation liquid at the same time, and the control group was used as the control group. No inflammatory reaction was observed in the group, indicating that the control group was not contaminated by the stain group. The LPS interfered with the pulmonary alveolar blood of the bacterial group. There was a significant difference in lung organ coefficient between LPS and control group (F = 5.699, P = 0. 006), and there was a significant difference between the time points (F = 10.648, P0.001), group and time. point-to-point interaction There was no significant difference (F = 0. 668, P0.05). Compared with the control group (P <0.05), the organ coefficients of the LPS-treated group and the LPS-free group were higher than those in the control group (P0.05). There was no significant difference in the level of TLR4 protein in the three groups (F = 47. 144, P 0.001), and the difference of time point was statistically significant (F = 141.543, P0.001), and the interaction between the group and time point was statistically significant (F = 141.543, P 0.001). The difference was also of statistical significance (F = 29. 902, P0. The expression level of TLR4 in LPS-treated group was significantly higher than that of control group (P <0.05). The expression of TLR4 in LPS-free group was higher than that of control group (P <0.05), and the level of TLR4 expression in LPS-free group was higher than that of control group (P0.05). The TLR4 protein in both group and LPS group reached the peak at 12h, and the level of TLR4 in LPS group was higher than that of LPS group (P0.05). There was no significant difference in the expression of TLR4 in the two groups. The difference between the three groups was statistically significant (F = 25. 817, P 0.001). The primary effect of time was also of statistical significance (F = 8.3933, P 0.001). There was no interaction between the group and the time point (F = I. 460, P0.05). There was a significant difference in the concentration of TNF-1 between the group and the LPS-treated group, and the levels of TNF-1 in the LPS-free group were higher than that of the LPS-treated group. The concentration of TNF-1 in the two groups was decreased after 24 h of infection. There was no significant difference in the concentration of IL-1 in the three groups (F = 18. 990, P. 001). The primary effect of time was not significant (F = I. 323, P0.05). There was no interaction between the group and the time point (F = 0.382, P0.05). After the LSD method, the levels of IL-1 in the LPS-treated group and the control group were not found to be statistically different, but the LPS did not interfere with the control group and the control group. There was no significant difference in the concentration of IL-1 in the group and LPS intervention group, and LPS did not interfere. bacteria The levels of IL-1 at all time points in the group were higher than that of the LPS-treated group. The expression level of TLR4 in the high-dose mice was significantly increased by LPS intervention and short-time effect. The concentration of TNF-1 and IL-1 in the group-infected cell group was higher than that of the control group. After the infection of Legionella, the expression level of TNF-1 and IL-1 was up-regulated in order to resist the infection of Legionella.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.1
【参考文献】
相关期刊论文 前10条
1 蒋志勤,阮先成;空调系统与军团菌感染因素探讨[J];湖北预防医学杂志;2001年05期
2 金建敏;军团病研究进展[J];国外医学(内科学分册);2001年10期
3 胡大林,廖建坤,杨光,欧阳小明;军团菌病[J];国外医学(卫生学分册);2003年04期
4 路凤;金银龙;程义斌;;军团菌病的流行概况[J];国外医学(卫生学分册);2008年02期
5 杨一新;李桂源;;LPS所介导的信号转导通路研究进展[J];中南大学学报(医学版);2006年01期
6 邵祝军;军团菌病的监测与防治[J];疾病监测;2005年06期
7 王刚毅,陈悦,沈健民,张曦,姜庆五,林海江;呼吸道感染病人军团菌抗体水平调查[J];疾病控制杂志;2005年01期
8 康晓明,汤忠群,夏锡荣;嗜肺军团菌感染1例报告[J];解放军医学杂志;1982年04期
9 郭菲,汪泱,方方,邢娟娟,彭燕,黄学明;脂多糖刺激后腹腔巨噬细胞表面Toll样受体表达的变化[J];江西医学检验;2004年06期
10 杨占清,刘运喜,彭清平,王宏伟,王闵,彭佐林,于晓敏,冯茂全,索翠萍,芮庆广;山东部分地区军团杆菌感染临床流行病学调查分析[J];实用医药杂志;2002年05期
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