本文選題：COPD　切入點：S100A8　出處：《川北醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:探討鈣結(jié)合蛋白S100A8、S100A9在慢性阻塞性肺疾病(COPD)大鼠肺泡巨噬細胞中的表達及其作用。方法:1.將12只健康成年雄性大鼠隨機分為兩組:COPD組和正常組。采用煙熏加氣管內(nèi)注射內(nèi)毒素方法1個月建立COPD大鼠模型。顯微鏡下觀察兩組大鼠肺組織病理形態(tài),圖像分析軟件測定肺平均內(nèi)襯間隔(MLI)、平均肺泡數(shù)(MAN)、肺泡腔面積與總面積比(PAA),瑞氏染色法檢測兩組大鼠支氣管肺泡灌洗液(BLAF)中細胞總數(shù)、肺泡巨噬細胞、淋巴細胞、中性粒細胞的絕對值和比例,分離、培養(yǎng)COPD組和正常組大鼠肺泡巨噬細胞,分別予以不同劑量S100A8、S100A9處理6h、12h。采用酶聯(lián)免疫吸附法(ELSA)檢測大鼠肺泡巨噬細胞上清液中IL-6、IL-8和TNF-a的濃度,原位雜交方法和免疫組化方法觀察離體培養(yǎng)的大鼠肺泡巨噬細胞中S100A8、S100A9mRNA和S100A8/A9蛋白的表達情況。結(jié)果:(1)與正常對照組比較,COPD組大鼠肺組織出現(xiàn)了典型的COPD病理改變,且MLI、PAA比正常對照組明顯增高(P0.05),而MAN則顯著低于正常對照組(P0.05)。(2)COPD組BALF中細胞總數(shù)與各炎癥細胞的絕對值較正常對照組顯著增高(P均0.05),其中,肺泡巨噬細胞絕對值升高最為明顯。(3)原位雜交顯示S100A8、S100A9mRNA主要表達在大鼠肺泡巨噬細胞細胞質(zhì)與細胞膜,統(tǒng)計分析示COPD組大鼠肺泡巨噬細胞中S100A8、S100A9mRNA表達較正常對照組顯著增加(P0.05)。(4)免疫組化顯示S100A8/A9蛋白也主要表達在大鼠肺泡巨噬細胞細胞質(zhì)和胞膜,統(tǒng)計分析示COPD組大鼠肺泡巨噬細胞中S100A8/A9蛋白表達量亦較正常對照組明顯增加(P0.05))。(5)ELISA結(jié)果顯示:S100A8、S100A9刺激兩組大鼠肺泡巨噬細胞釋放炎癥因子IL-6、IL-8、TNF-α呈劑量和時間依賴性增加。在相同濃度的S100A8、S100A9作用相同時間時,COPD組大鼠肺泡巨噬細胞上清液中IL-6、IL-8、TNF-α的濃度均高于正常對照組(P均0.05)。相同劑量S100A8與S100A9作用大鼠肺泡巨噬細胞比較,S100A8處理兩組大鼠肺泡巨噬細胞后上清液中IL-8和TNF-α的濃度均高于S100A9處理兩組大鼠肺泡巨噬細胞后上清液中IL-8和TNF-α的濃度(P均0.05);而較高劑量S100A8刺激COPD及正常大鼠肺泡巨噬細胞分泌IL-6的量也均高于S100A9刺激兩組大鼠肺泡巨噬細胞分泌IL-6的量(P均0.05)。結(jié)論:(1)COPD大鼠模型可以通過煙熏加氣管內(nèi)注射內(nèi)毒素方法建成。(2)COPD大鼠肺泡巨噬細胞內(nèi)S100A8、S100A9mRNA以及S100A8/A9蛋白的表達量較正常對照組明顯增加。(3)S100A8、S100A9呈時間、濃度依賴性刺激大鼠肺泡巨噬細胞分泌炎癥因子IL-6、IL-8和TNF-α增多。(4)S100A8、S100A9刺激COPD大鼠肺泡巨噬細胞分泌IL-6、IL-8和TNF-α增多作用較正常大鼠肺泡巨噬細胞更明顯。(5)S100A8刺激大鼠肺泡巨噬細胞分泌IL-6、IL-8和TNF-α增多作用明顯強于S100A9。
[Abstract]:Objective: to investigate the expression and role of calcium binding protein S100A8 (S100A8) S100A9 in alveolar macrophages of rats with chronic obstructive pulmonary disease (COPD). Methods: 1. Twelve healthy adult male rats were randomly divided into two groups: the control group and the control group. COPD rat model was established by endotoxin injection into trachea for 1 month. The pathological morphology of lung tissue was observed under microscope in two groups. The image analysis software was used to measure the mean lining interval (MLI), the average number of alveoli (MAN), the ratio of alveolar cavity area to total area (PAA). The total number of cells, alveolar macrophages and lymphocytes in bronchoalveolar lavage fluid (BLAF) of the two groups were detected by Rayleigh staining. The absolute value and proportion of neutrophils, isolation and culture of alveolar macrophages in COPD group and normal group were treated with different doses of S100A8 / S100A9 for 6 h and 12 h respectively. The concentrations of IL-6IL-8 and TNF-a in the supernatant of rat alveolar macrophages were detected by enzyme-linked immunosorbent assay (Elisa). The expression of S100A8, S100A9 mRNA and S100A9 protein in cultured rat alveolar macrophages was observed by in situ hybridization and immunohistochemistry. Compared with the normal control group, the total number of BALF cells and the absolute values of the inflammatory cells in the MAN group were significantly higher than those in the normal control group (P 0.05), especially in the normal control group (P < 0.05), and the total number of BALF cells and the absolute values of the inflammatory cells in the BALF group were significantly higher than those in the normal control group (P < 0.05). In situ hybridization showed that S100A8 and S100A9 mRNA were mainly expressed in the cytoplasm and cell membrane of alveolar macrophages in rats. Statistical analysis showed that the expression of S100A8, S100A9 mRNA in alveolar macrophages in COPD group was significantly higher than that in normal control group (P 0.05, P 0.05, P < 0.05). Immunohistochemical staining showed that S100A8/A9 protein was also mainly expressed in the cytoplasm and membrane of alveolar macrophages in rats. Statistical analysis showed that the expression of S100A8/A9 protein in alveolar macrophages in COPD group was also significantly higher than that in normal control group. The results of Elisa showed that the inflammatory factor IL-6 and IL-8 TNF- 偽 were increased in a dose-and time-dependent manner after stimulated by S100A8 and S100A9 in two groups. At the same time, the concentration of IL-6 and IL-8 TNF- 偽 in alveolar macrophage supernatant of COPD group was higher than that of normal control group (P < 0.05). The comparison of alveolar macrophages treated with S100A8 and S100A9 in the same dose of S100A8 and S100A9 was made. The concentrations of IL-8 and TNF- 偽 in supernatants of alveolar macrophages were higher than those of IL-8 and TNF- 偽 in the supernatants of alveolar macrophages treated with S100A9 in rats (P < 0.05), while the concentrations of IL-6 secreted by COPD and normal alveolar macrophages were stimulated by high dose S100A8. The amount of IL-6 secreted by alveolar macrophages stimulated by S100A9 was also higher than that of both groups (P < 0.05). Conclusion the S100A8 S100A9 mRNA and the surface of S100A8/A9 protein in alveolar macrophages can be established by smoking and endotoxin injection into the alveolar macrophages. Compared with the normal control group, the volume of S100A8 and S100A9 increased significantly. In a dose-dependent manner, the cytokines IL-6, IL-8 and TNF- 偽 from alveolar macrophages were stimulated in a dose-dependent manner. S100A8 and S100A9 stimulated the production of IL-6, IL-8 and TNF- 偽 by alveolar macrophages in COPD rats, which were more obvious than those in normal rat alveolar macrophages. 5S100A8 stimulated alveolar macrophages in rats. The increase of IL-6, IL-8 and TNF- 偽 was stronger than that of S100A9.
【學(xué)位授予單位】：川北醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R563.9

【參考文獻】

相關(guān)期刊論文 前7條

1 邱貞琴;張華;;蘿卜硫素對慢阻肺患者TLR4、MyD88以及下游炎性因子的影響分析[J];臨床肺科雜志;2016年03期

2 張宇鵬;;烏司他丁治療對COPD急性加重期患者血清TLRs、MMPs以及脂質(zhì)過氧化的影響[J];海南醫(yī)學(xué)院學(xué)報;2015年02期

3 ;慢性阻塞性肺疾病診治指南(2013年修訂版)[J];中國醫(yī)學(xué)前沿雜志(電子版);2014年02期

4 支修益;吳一龍;馬勝林;王天佑;王長利;王潔;石遠凱;盧鈾;劉倫旭;劉德若;陳東紅;楊躍;杜祥;步宏;周清華;姜格寧;韓寶惠;程剛;程穎;焦順昌;;原發(fā)性肺癌診療規(guī)范(2011年版)[J];中國肺癌雜志;2012年12期

5 顧延會;歐陽瑤;;煙熏聯(lián)合脂多糖制備大鼠慢性阻塞性肺疾病動物模型[J];重慶醫(yī)學(xué);2012年13期

6 尹磊淼;張慶華;王宇;徐玉東;楊永清;;鈣結(jié)合蛋白S100A9功能的研究進展[J];復(fù)旦學(xué)報(醫(yī)學(xué)版);2011年05期

7 金焱,龐寶森,武維屏,馮淬靈,任傳云,牛淑潔,黃秀霞,崔巍,阮英茆;一種實驗性大鼠慢性阻塞性肺疾病模型的建立[J];心肺血管病雜志;2004年03期

，

本文編號：1675098