發(fā)布時(shí)間：2018-06-09 06:34

  本文選題：矽肺 + 肺泡巨噬細(xì)胞　； 參考：《華北理工大學(xué)》2017年碩士論文

【摘要】：目的探討矽肺患者肺泡巨噬細(xì)胞中脂質(zhì)代謝的變化,分析其對(duì)矽肺纖維化發(fā)生發(fā)展的作用,進(jìn)一步闡明矽肺發(fā)生的機(jī)制,為矽肺治療提供新思路。方法研究對(duì)象選取:研究對(duì)象為北戴河中國(guó)煤礦工人療養(yǎng)院的首次進(jìn)行肺灌洗手術(shù)的矽肺患者,主要從事開(kāi)拓、掘進(jìn)、粉碎、風(fēng)鉆、石材加工等工作。由塵肺診斷專家組依據(jù)GBZ-70-2009《塵肺病診斷標(biāo)準(zhǔn)》對(duì)患者矽肺期別進(jìn)行集體診斷。其中,矽肺壹期18例,矽肺貳期17例,矽肺叁期24例,均為男性,對(duì)照組8例,包括5例矽肺觀察對(duì)象,3例誤吸患者。此外,選取12例健康成年男性作為血清對(duì)照組。納入研究對(duì)象時(shí),排除患有傳染性疾病、肺部疾病如肺炎、肺心病、活動(dòng)期肺結(jié)核及患有其他部位纖維化的矽肺患者,同時(shí)排除肝功能異常和高脂血癥患者。對(duì)照組、血清對(duì)照組、矽肺壹期、矽肺貳期、矽肺叁期患者的年齡、體質(zhì)指數(shù)(BMI)比較差異無(wú)顯著性。生物樣本收集:抽取研究對(duì)象空腹外周靜脈血3ml,離心提取血清樣本;收集研究對(duì)象適量肺泡灌洗回收液,提取其中的肺泡巨噬細(xì)胞,保留上清液,所有樣本均于-80℃保存。實(shí)驗(yàn)方法:應(yīng)用HE、油紅O染色對(duì)巨噬細(xì)胞進(jìn)行形態(tài)學(xué)觀察;應(yīng)用透射電鏡觀察巨噬細(xì)胞超微結(jié)構(gòu)。應(yīng)用薄層色譜法(TLC)對(duì)肺泡巨噬細(xì)胞及肺泡灌洗液中的脂質(zhì)進(jìn)行分離定性;采用雙抗體夾心法(ELISA)測(cè)定肺泡巨噬細(xì)胞及肺泡灌洗液中轉(zhuǎn)化因子β(TGF-β1)、結(jié)締組織生長(zhǎng)因子(CTGF)和氧化低密度脂蛋白(ox-LDL)濃度,同時(shí)測(cè)定血清TGF-β1、CTGF濃度;比色法測(cè)定肺泡巨噬細(xì)胞及肺泡灌洗液中膽固醇、甘油三酯、磷脂濃度。結(jié)果1 HE染色發(fā)現(xiàn),與對(duì)照組相比,矽肺各組肺泡巨噬細(xì)胞胞質(zhì)上出現(xiàn)大量空泡,胞漿中含大量吞噬顆粒,以叁期矽肺最為明顯。2油紅O染色發(fā)現(xiàn),與對(duì)照組相比,矽肺各組肺泡巨噬細(xì)胞中出現(xiàn)橘紅色脂滴,胞漿中可見(jiàn)大量吞噬顆粒,以叁期矽肺最為明顯,確定肺泡巨噬細(xì)胞中存在脂質(zhì)。3電鏡發(fā)現(xiàn)矽肺各組肺泡巨噬細(xì)胞出現(xiàn)偽足斷裂、消失,細(xì)核染色質(zhì)邊集,染色加深;次級(jí)溶酶體增加,線粒體發(fā)生腫脹、固縮、空泡,胞質(zhì)中出現(xiàn)密度均一,大小不等的脂滴等現(xiàn)象,以矽肺貳期、叁期患者最為明顯。4薄層色譜結(jié)果顯示肺泡巨噬細(xì)胞和肺泡灌洗液中含有膽固醇、甘油三酯、磷脂三種脂質(zhì)。5隨著矽肺分期增高,AM和BALF中膽固醇含量均呈上升趨勢(shì),矽肺叁期組均明顯高于對(duì)照組、矽肺壹期組、矽肺貳期組,差異具有顯著性(P0.05);AM中ox-LDL和磷脂水平隨矽肺分期增高呈上升趨勢(shì),而B(niǎo)ALF中ox-LDL水平呈下降趨勢(shì),各組間兩兩比較差異均具有顯著性(P0.05),BALF中的磷脂水平在各組間兩兩比較差異也均具有顯著性(P0.05),其中矽肺各組均明顯高于對(duì)照組,矽肺貳期組明顯低于矽肺壹期組,矽肺叁期組明顯高于矽肺壹期組、矽肺貳期組。在各組BALF中,甘油三酯水平的差別不具有統(tǒng)計(jì)學(xué)意義(P0.05)。6血清中TGF-β、CTGF水平隨矽肺進(jìn)展呈上升趨勢(shì),組間兩兩比較,差異均具有顯著性(P0.05),且血TGF-β水平與血CTGF水平呈正相關(guān)(P0.05);對(duì)照組肺泡巨噬細(xì)胞中TGF-β、CTGF水平均明顯低于矽肺各組(P0.05),且呈先上升后下降的趨勢(shì),TGF-β、CTGF水平分別在矽肺壹期、矽肺貳期達(dá)到最大值;BALF中各組TGF-β、CTGF水平差異不具有統(tǒng)計(jì)學(xué)意義(P0.05)。7 AM中膽固醇、甘油三酯、磷脂、ox-LDL水平與血CTGF、TGF-β水平呈正相關(guān)(P0.05),BALF中膽固醇、磷脂水平與血CTGF、TGF-β水平呈正相關(guān)(P0.05),BALF中ox-LDL水平與血CTGF、TGF-β水平呈負(fù)相關(guān)(P0.05),BALF中甘油三酯水平與血CTGF、TGF-β水平不存在相關(guān)關(guān)系(P0.05)。結(jié)論1矽肺患者肺泡巨噬細(xì)胞發(fā)生泡沫化,存在脂質(zhì)蓄積的現(xiàn)象。2隨著矽肺纖維化的發(fā)展,矽肺患者血清中CTGF、TGF-β水平呈上升趨勢(shì),肺泡巨噬細(xì)胞中CTGF、TGF-β水平呈先升高后降低的趨勢(shì),且分別在矽肺壹期、貳期達(dá)最大值。3在矽肺纖維化發(fā)展進(jìn)程中,肺泡巨噬細(xì)胞存在脂質(zhì)代謝失衡的現(xiàn)象,且肺泡巨噬細(xì)胞中的脂質(zhì)蓄積量隨著纖維化程度的加重呈上升趨勢(shì),其機(jī)制可能與巨噬細(xì)胞發(fā)生泡沫化有關(guān)。
[Abstract]:Objective to investigate the changes of lipid metabolism in alveolar macrophages in silicosis patients and to analyze its effect on the development of silicosis and further elucidate the mechanism of silicosis to provide new ideas for the treatment of silicosis. Methods the object of study was to study the silicosis of lung lavage operation for the first time in Beidaihe Chinese coal mine workers' sanatorium. Patients, mainly engaged in the work of opening, digging, crushing, wind drilling, stone processing, and so on. The pneumoconiosis expert group, according to the diagnostic criteria of GBZ-70-2009< pneumoconiosis, has a group diagnosis of silicosis. Among them, 18 cases of silicosis, 17 silicosis stage II, 24 silicosis, 8 cases of the control group, 5 cases of silicosis and 3 cases. In addition, 12 healthy adult men were selected as a serum control group. The subjects were excluded from infectious diseases, pulmonary diseases such as pneumonia, pulmonary heart disease, pulmonary tuberculosis in the active stage and silicosis patients with other parts of fibrosis. Meanwhile, patients with liver dysfunction and hyperlipidemia were excluded. Control group, serum control group, silicosis one. There was no significant difference in age and body mass index (BMI) in patients with silicosis. Biological samples were collected to extract 3ml from fasting peripheral venous blood and centrifugally extract serum samples; collect the appropriate alveolar lavage recovery solution, extract the alveolar macrophages, and retain the supernatant, all samples were kept at -80 C. Experimental methods: HE, oil red O staining was used to observe the morphology of macrophages; the ultrastructure of macrophages was observed by transmission electron microscopy. The lipid of alveolar macrophages and alveolar lavage fluid was separated and qualitative by TLC (TLC); and the double antibody sandwich (ELISA) method was used to determine the alveolar macrophage and alveolar lavage solution. Transformation factor beta (TGF- beta 1), connective tissue growth factor (CTGF) and oxidized low density lipoprotein (ox-LDL) concentration, serum TGF- beta 1, CTGF concentration, cholesterol, triglyceride, and phospholipid concentration in alveolar macrophages and alveolar lavage fluid were measured by colorimetry. Results 1 HE staining was found, compared with the control group, alveolar macrophage cells in each group were compared with the control group. A large number of vacuoles were found in the cytoplasm, and a large number of phagocytic particles were contained in the cytoplasm. The most obvious.2 oil red O staining was found in the three phases of silicosis. Compared with the control group, there were orange red drops in the alveolar macrophages in each group of silicosis and a large number of phagocytic particles in the cytoplasm, and the most obvious three phases of silicosis were found in three stages of silicosis. The lipid.3 electron microscopy found silicosis in the alveolar macrophages. Pulmonary alveolar macrophages in all pulmonary alveolar macrophages appeared to be false foot fracture, disappeared, chromatin set and dyed in fine nucleus, secondary lysosome increased, mitochondria were swelling, pyknosis, vacuoles, and lipid droplets with uniform density and different sizes appeared in the cytoplasm. The most obvious.4 TLC results showed alveolar macrophages and alveoli in three phase patients. The contents of cholesterol, triglyceride and phospholipid of three kinds of lipid.5 in the lavage fluid increased with the increase of silicosis, and the levels of cholesterol in AM and BALF were all higher than those in the control group. The difference was significant (P0.05) in the one stage Silicosis group and the silicosis stage group (P0.05), and the level of ox-LDL and phospholipid in AM increased with the increase of the stage of silicosis. The level of ox-LDL in BALF showed a downward trend, and the difference of 22 in each group was significant (P0.05). The level of phospholipid in BALF was significantly higher than that of the control group (P0.05). The Silicosis group was significantly higher than the control group, and the silicosis II group was significantly lower than the one stage Silicosis group, and the Silicosis group was significantly higher than the silicosis one group. The difference of triglyceride level in each group of BALF was not statistically significant (P0.05).6 serum TGF- beta, CTGF level increased with the progress of silicosis, and the difference between the 22 groups was significant (P0.05), and the level of TGF- beta in blood was positively correlated with the level of blood CTGF (P0.05); TGF- beta, CTGF water in the alveolar macrophages in the control group. The average value was significantly lower than that of silicosis (P0.05), and the level of TGF- beta, CTGF was in the first phase of silicosis, and the stage of silicosis reached the maximum, and the difference of TGF- beta and CTGF levels in BALF was not statistically significant (P0.05) in.7 AM, the level of glycerol three ester, phospholipid, ox-LDL was positively related to the CTGF of blood, and the TGF- beta level was positively correlated. There was a positive correlation between the level of cholesterol and phospholipid in BALF and the level of CTGF and TGF- beta in blood (P0.05). The level of ox-LDL in BALF was negatively correlated with the level of CTGF and TGF- beta in blood (P0.05). There was no correlation between the level of triglyceride in BALF and the CTGF of blood and the level of TGF- beta. Conclusion the alveolar macrophages in 1 silicosis patients were foamed, and the accumulation of lipids existed with them. The development of silicosis fibrosis, the level of CTGF and TGF- beta in the serum of silicosis patients increased. The level of CTGF and TGF- beta in alveolar macrophages showed a tendency to increase first and then decrease, and the maximum value of.3 in silicosis was respectively at.3 in the development of silicosis, and the imbalance of lipid metabolism in the alveolar macrophage cells, and the alveolar macrophage was fine. The accumulation of lipid in the cells showed an upward trend with the increase of fibrosis. The mechanism may be related to the foaming of macrophages.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R135.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 彭哲;周蕓;張海;夏穎;史廷明;劉躍偉;;轉(zhuǎn)化生長(zhǎng)因子與結(jié)締組織生長(zhǎng)因子在石英致肺纖維化中的作用[J];公共衛(wèi)生與預(yù)防醫(yī)學(xué);2016年04期

2 吳傳文;張秀峰;劉微;王宏麗;郝小惠;郭志義;郭靈麗;劉和亮;;表面活性蛋白-Drs3088308、rs721917位點(diǎn)基因多態(tài)性與矽肺易感性的關(guān)系[J];南方醫(yī)科大學(xué)學(xué)報(bào);2016年07期

3 趙金垣;王世俊;;塵肺應(yīng)為可治之癥[J];環(huán)境與職業(yè)醫(yī)學(xué);2016年01期

4 曹華;馬紅英;曹霞;黃新莉;張雪靜;周曉紅;;外源性H_2S對(duì)肺纖維化大鼠肺組織轉(zhuǎn)化生長(zhǎng)因子-β1及結(jié)締組織生長(zhǎng)因子的干預(yù)作用[J];中國(guó)老年學(xué)雜志;2014年01期

5 馬文東;袁媛;楊奕;徐洪;鄧海靜;于婉瑩;孫月;魏中秋;楊方;;TGF-β_1介導(dǎo)的RhoA/ROCK通路在大鼠肺肌成纖維細(xì)胞分化中的調(diào)節(jié)作用[J];中國(guó)病理生理雜志;2013年10期

6 叢翠翠;毛麗君;趙金垣;;肺淋巴系統(tǒng)與塵肺[J];中國(guó)工業(yè)醫(yī)學(xué)雜志;2013年03期

7 趙國(guó)軍;湯石林;田國(guó)平;歐陽(yáng)新平;呂運(yùn)成;何平平;姚峰;唐艷艷;吳劍鋒;王甲林;張敏;唐朝克;;PDTC對(duì)ox-LDL誘導(dǎo)的THP-1巨噬細(xì)胞脂質(zhì)蓄積和膽固醇流出的影響[J];中南醫(yī)學(xué)科學(xué)雜志;2013年03期

8 陳冕;何曉曉;石碧華;王柯敏;陳素葉;周冰;;不同尺寸二氧化硅納米顆粒體內(nèi)分布與代謝研究[J];科學(xué)通報(bào);2013年07期

9 趙靜;吳秀珍;王獻(xiàn)華;;經(jīng)SiO_2粉塵刺激的矽肺患者肺泡巨噬細(xì)胞中CTGF的表達(dá)及意義[J];現(xiàn)代預(yù)防醫(yī)學(xué);2011年11期

10 張弛;黃靚;佘美華;尹衛(wèi)東;;褪黑素對(duì)ox-LDL所致巨噬細(xì)胞TNF-α釋放及核因子κB活性的影響[J];中南醫(yī)學(xué)科學(xué)雜志;2011年01期

相關(guān)博士學(xué)位論文 前1條

1 周勇;環(huán)氧二十碳三烯酸對(duì)急性肺損傷保護(hù)作用的初步研究[D];中南大學(xué);2013年

相關(guān)碩士學(xué)位論文 前3條

1 劉微;肺淋巴循環(huán)在矽肺發(fā)病機(jī)制中的作用研究[D];華北理工大學(xué);2016年

2 陳慧;接矽塵作業(yè)人員體內(nèi)硅元素水平變化及與TGF-β1、TNF-α含量關(guān)系的研究[D];河北聯(lián)合大學(xué);2014年

3 劉梁英;川芎嗪、大黃酸聯(lián)用對(duì)轉(zhuǎn)化生長(zhǎng)因子β_1和甘油三酯致肝細(xì)胞損傷的影響[D];重慶醫(yī)科大學(xué);2005年

，

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