矽肺患者肺泡巨噬细胞中脂质代谢的变化

发布时间：2018-06-09 06:34

本文选题：矽肺 + 肺泡巨噬细胞　；参考：《华北理工大学》2017年硕士论文

【摘要】：目的探讨矽肺患者肺泡巨噬细胞中脂质代谢的变化,分析其对矽肺纤维化发生发展的作用,进一步阐明矽肺发生的机制,为矽肺治疗提供新思路。方法研究对象选取:研究对象为北戴河中国煤矿工人疗养院的首次进行肺灌洗手术的矽肺患者,主要从事开拓、掘进、粉碎、风钻、石材加工等工作。由尘肺诊断专家组依据GBZ-70-2009《尘肺病诊断标准》对患者矽肺期别进行集体诊断。其中,矽肺壹期18例,矽肺贰期17例,矽肺叁期24例,均为男性,对照组8例,包括5例矽肺观察对象,3例误吸患者。此外,选取12例健康成年男性作为血清对照组。纳入研究对象时,排除患有传染性疾病、肺部疾病如肺炎、肺心病、活动期肺结核及患有其他部位纤维化的矽肺患者,同时排除肝功能异常和高脂血症患者。对照组、血清对照组、矽肺壹期、矽肺贰期、矽肺叁期患者的年龄、体质指数(BMI)比较差异无显著性。生物样本收集:抽取研究对象空腹外周静脉血3ml,离心提取血清样本;收集研究对象适量肺泡灌洗回收液,提取其中的肺泡巨噬细胞,保留上清液,所有样本均于-80℃保存。实验方法:应用HE、油红O染色对巨噬细胞进行形态学观察;应用透射电镜观察巨噬细胞超微结构。应用薄层色谱法(TLC)对肺泡巨噬细胞及肺泡灌洗液中的脂质进行分离定性;采用双抗体夹心法(ELISA)测定肺泡巨噬细胞及肺泡灌洗液中转化因子β(TGF-β1)、结缔组织生长因子(CTGF)和氧化低密度脂蛋白(ox-LDL)浓度,同时测定血清TGF-β1、CTGF浓度;比色法测定肺泡巨噬细胞及肺泡灌洗液中胆固醇、甘油三酯、磷脂浓度。结果1 HE染色发现,与对照组相比,矽肺各组肺泡巨噬细胞胞质上出现大量空泡,胞浆中含大量吞噬颗粒,以叁期矽肺最为明显。2油红O染色发现,与对照组相比,矽肺各组肺泡巨噬细胞中出现橘红色脂滴,胞浆中可见大量吞噬颗粒,以叁期矽肺最为明显,确定肺泡巨噬细胞中存在脂质。3电镜发现矽肺各组肺泡巨噬细胞出现伪足断裂、消失,细核染色质边集,染色加深;次级溶酶体增加,线粒体发生肿胀、固缩、空泡,胞质中出现密度均一,大小不等的脂滴等现象,以矽肺贰期、叁期患者最为明显。4薄层色谱结果显示肺泡巨噬细胞和肺泡灌洗液中含有胆固醇、甘油三酯、磷脂三种脂质。5随着矽肺分期增高,AM和BALF中胆固醇含量均呈上升趋势,矽肺叁期组均明显高于对照组、矽肺壹期组、矽肺贰期组,差异具有显著性(P0.05);AM中ox-LDL和磷脂水平随矽肺分期增高呈上升趋势,而BALF中ox-LDL水平呈下降趋势,各组间两两比较差异均具有显著性(P0.05),BALF中的磷脂水平在各组间两两比较差异也均具有显著性(P0.05),其中矽肺各组均明显高于对照组,矽肺贰期组明显低于矽肺壹期组,矽肺叁期组明显高于矽肺壹期组、矽肺贰期组。在各组BALF中,甘油三酯水平的差别不具有统计学意义(P0.05)。6血清中TGF-β、CTGF水平随矽肺进展呈上升趋势,组间两两比较,差异均具有显著性(P0.05),且血TGF-β水平与血CTGF水平呈正相关(P0.05);对照组肺泡巨噬细胞中TGF-β、CTGF水平均明显低于矽肺各组(P0.05),且呈先上升后下降的趋势,TGF-β、CTGF水平分别在矽肺壹期、矽肺贰期达到最大值;BALF中各组TGF-β、CTGF水平差异不具有统计学意义(P0.05)。7 AM中胆固醇、甘油三酯、磷脂、ox-LDL水平与血CTGF、TGF-β水平呈正相关(P0.05),BALF中胆固醇、磷脂水平与血CTGF、TGF-β水平呈正相关(P0.05),BALF中ox-LDL水平与血CTGF、TGF-β水平呈负相关(P0.05),BALF中甘油三酯水平与血CTGF、TGF-β水平不存在相关关系(P0.05)。结论1矽肺患者肺泡巨噬细胞发生泡沫化,存在脂质蓄积的现象。2随着矽肺纤维化的发展,矽肺患者血清中CTGF、TGF-β水平呈上升趋势,肺泡巨噬细胞中CTGF、TGF-β水平呈先升高后降低的趋势,且分别在矽肺壹期、贰期达最大值。3在矽肺纤维化发展进程中,肺泡巨噬细胞存在脂质代谢失衡的现象,且肺泡巨噬细胞中的脂质蓄积量随着纤维化程度的加重呈上升趋势,其机制可能与巨噬细胞发生泡沫化有关。
[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.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R135.2

【参考文献】