本文關(guān)鍵詞：納米二氧化硅刺激BEAS-2B細(xì)胞炎癥和纖維化反應(yīng)及其與外周血相關(guān)細(xì)胞因子的關(guān)系研究　出處：《昆明醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 肺癌 炎癥反應(yīng) 纖維化 細(xì)胞因子 人支氣管上皮細(xì)胞 納米二氧化硅

【摘要】：背景與目的：肺癌是發(fā)生于支氣管粘膜上皮的惡性腫瘤。肺癌的病因至今尚不明確。云南省是我國(guó)肺癌的高發(fā)區(qū),特別是宣威地區(qū)的女性肺癌發(fā)病率居世界首位,前期相關(guān)研究顯示宣威地區(qū)肺癌的高發(fā)可能與燃煤中含有大量的納米二氧化硅顆粒,導(dǎo)致人吸入煙塵后肺部出現(xiàn)炎癥反應(yīng)有關(guān)。參與肺部炎癥反應(yīng)的細(xì)胞因子可分為促腫瘤類因子：IL-6、IL-8、IL-17、RANTES和MIP-1a;抗腫瘤類因子：IL-4、IL-13、IL-2、TNF-a, IFN-γ;促進(jìn)血管生成及纖維化類因子：VEGF、b-FGF, MCP-1、IL-12、IL-5。還有比較特殊的有IL-10,其對(duì)腫瘤的發(fā)生發(fā)展具有兩面性,既有利于腫瘤的生存又能抑制腫瘤生長(zhǎng)。本研究探討納米二氧化硅與人正常支氣管上皮細(xì)胞(BEAS-2B細(xì)胞)的關(guān)系,探索炎癥反應(yīng)在肺癌發(fā)生和進(jìn)展中的作用,為腫瘤尋找合適的標(biāo)記物奠定基礎(chǔ)。方法：1、分組采用體外細(xì)胞培養(yǎng)的方式,在1640培養(yǎng)基中加入不同粒徑納米二氧化硅,通過(guò)不同粒徑納米二氧化硅刺激人正常支氣管上皮細(xì)胞(BEAS-2B細(xì)胞),并于24h后觀察其形態(tài)學(xué)大小、細(xì)胞結(jié)構(gòu)的改變；濃度為10μg/mL、25μg/mL、50ug/mL進(jìn)行實(shí)驗(yàn),一次性刺激24h后,經(jīng)過(guò)離心后取上層清液。2、采用Bio-Plex懸液芯片系統(tǒng)檢測(cè)納米二氧化硅刺激BEAS-2B細(xì)胞24h后培養(yǎng)上清液中促腫瘤類因子：IL-6、IL-8、IL-17、RANTES和MIP-1a;抗腫瘤類因子：IL-4、IL-13、IL-2、TNF-a、IFN-γ;促進(jìn)血管生成及纖維化類因子：VEGF、 b-FGF,MCP-1、IL-12、IL-5;同時(shí)具有促腫瘤及抗腫瘤作用炎性因子IL-10的水平變化。3、收集宣威地區(qū)女性非小細(xì)胞肺癌(NSCLC)患者、非宣威地區(qū)女性非小細(xì)胞肺癌患者外周血各20例、健康女性的外周血20例,按照不同臨床分期、病理類型、分化程度、淋巴結(jié)轉(zhuǎn)移進(jìn)行統(tǒng)計(jì),經(jīng)過(guò)離心后取上層血清。采用Bio-Plex懸液芯片系統(tǒng)檢測(cè)血清中促腫瘤了類因子:IL-6、IL-8、IL-17、RANTES和MIP-1a;抗腫瘤類因子：IL-4、IL-13、IL-2、TNF-a、IFN-γ:促進(jìn)血管生成及纖維化類因子：VEGF、b-FGF, MCP-1、IL-12、IL-5:同時(shí)具有促腫瘤及抗腫瘤作用炎性因子IL-10的水平變化。結(jié)果：1、納米二氧化硅刺激下BEAS-2B細(xì)胞形態(tài)學(xué)改變：納米二氧化硅刺激細(xì)胞24h后,與對(duì)照組相比,在鏡下觀察可見(jiàn)實(shí)驗(yàn)組細(xì)胞生長(zhǎng)狀態(tài)欠佳,細(xì)胞體積增大,部分細(xì)胞胞質(zhì)內(nèi)可見(jiàn)空泡樣改變,細(xì)胞邊緣變鈍、不規(guī)則,50ug/mL濃度下部分細(xì)胞出現(xiàn)漂浮。2、納米二氧化硅致BEAS-2B細(xì)胞炎性反應(yīng)：各濃度刺激下,不同粒徑的納米二氧化硅可引起B(yǎng)EAS-2B細(xì)胞上清液中抗腫瘤類因子：RANTES因子在30nm組與50nm組之間存在差異性(P0.05)。促進(jìn)血管生成及纖維化類因子：、EGF因子在對(duì)照組與30nm組之間存在差異性(P0.05),IL-12因子在對(duì)照組與30nm、 50nm組之間存在差異性(P0.05)；同時(shí)具有促腫瘤及抗腫瘤作用炎性因子IL-10在對(duì)照組、30nm組、50nm組各組之間均存在差異性(P0.05)。3、宣威地區(qū)女性肺癌外周血：宣威地區(qū)女性肺癌患者、非宣威地區(qū)女性肺癌患者以及健康女性血清中抗腫瘤類因子：IL-6因子在正常組與宣威組之間存在顯著差異性(P0.01),IL17因子在正常組與宣威組、非宣威組之間存在差異性(P0.05)。抗腫瘤類因子：IL-4因子在正常組與非宣威組之間存在差異性(P0.05),IL-13在正常組與宣威組、非宣威組之間存在顯著差異性(P0.01),IFN-γ在正常組與宣威組之間存在差異性(P0.05);促進(jìn)血管生成及纖維化類因子：VEGF因子在正常組與宣威組、非宣威組之間存在顯著差異性(P0.01),b-FGF因子在正常組與非宣威組之間存在差異性(P0.05),IL-12因子在正常組與宣威組、非宣威組之間存在顯著差異性(P0.01)；同時(shí)具有促腫瘤及抗腫瘤作用炎性因子IL-10在正常組與宣威組之間存在差異性(P0.05)、在正常組與非宣威組之間存在顯著差異性(P0.01)。結(jié)論：1、納米二氧化硅對(duì)人支氣管上皮細(xì)胞有刺激作用,能促進(jìn)支氣管上皮發(fā)生重度不典型增生,可能與肺癌發(fā)生有關(guān)。2、通過(guò)檢測(cè)炎癥相關(guān)及纖維化相關(guān)細(xì)胞因子的分泌狀態(tài)可在一定程度上提示患者病情變化。3、宣威女性肺癌患者外周血存在差異性表達(dá)的細(xì)胞因子,可能與云南宣威女性肺癌的高發(fā)原因有關(guān)。
[Abstract]:Background and objective: lung cancer is occurring in bronchial epithelial malignant tumors. The etiology of lung cancer is still not clear. China's Yunnan province is a high incidence of lung cancer, especially in female lung cancer Xuanwei rate ranks first in the world, the research shows that Xuanwei lung cancer may contain a large number of nano silica particles and coal in the cause of inflammation of lung after smoke inhalation. Those cytokines involved in lung inflammation can be divided into types of tumor promoting factors: IL-6, IL-8, IL-17, RANTES and MIP-1a; antitumor factor: IL-4, IL-13, IL-2, TNF-a, IFN- gamma; promote angiogenesis and fibrosis cytokines: VEGF. B-FGF, MCP-1, IL-12, IL-5. and the special IL-10, with the two sides of the tumor development, is conducive to the survival of tumor and inhibit tumor growth. In this study, two nano oxide The silicon and the normal human bronchial epithelial cells (BEAS-2B cells) to explore the relationship of inflammation in lung cancer and its role in the progress, lay the foundation for appropriate markers for tumor. Methods: 1 groups using cell culture in vitro, in 1640 medium was added with particle size of nano silica by different particle the diameter of nano silica stimulation of normal human bronchial epithelial cells (BEAS-2B cells), and observe the morphology size after 24h, the change of cell structure; the concentration of 10 g/mL, 25 g/mL, 50ug/mL experiment, one-time after 24h stimulation, after centrifugation the supernatant liquid of.2, using Bio-Plex chip detection system of nano silica suspension stimulation of BEAS-2B cells after cultured 24h tumor promoting factor was: IL-6, IL-8, IL-17, RANTES and MIP-1a; antitumor factor: IL-4, IL-13, IL-2, TNF-a, IFN- gamma; promote angiogenesis and fiber Classes: VEGF, b-FGF, factor MCP-1, IL-12, IL-5;.3 levels also has tumor promoting and antitumor effects of inflammatory factor IL-10, collected the Xuanwei female non-small cell lung cancer (NSCLC) patients, female non Xuanwei patients with non-small cell lung cancer in peripheral blood of the 20 cases, 20 cases of peripheral the blood of healthy women, according to the different clinical stage, pathological type, differentiation degree, lymph node metastasis were analyzed by centrifugation after taking the serum. Using Bio-Plex suspension array system for detection of serum tumor promoting factor class: IL-6, IL-8, IL-17, RANTES and MIP-1a; antitumor factor: IL-4, IL-13, IL-2 TNF-a, IFN-, Gamma: promote angiogenesis and fibrosis cytokines: VEGF, b-FGF, MCP-1, IL-12, IL-5: levels also can promote tumor and anti-tumor effect of inflammatory factor IL-10. Results: 1, nano silica thorn morphology of BEAS-2B cell change: two nm excitation The silicon oxide stimulation of 24h cells, compared with control group, experimental group was observed under the microscope the growth state of poor cell volume increases, some cells in cytoplasm vacuolization, cell edge blunt, irregular, floating part of.2 cell 50ug/mL concentration, nano silica induced BEAS-2B cell inflammatory response the concentration under the stimulation of silica nanoparticles with different diameters can be induced by anti-tumor cytokines in the supernatants of BEAS-2B cells: RANTES factor differences between 30nm group and 50nm group (P0.05). To promote the angiogenesis and fibrosis factor: EGF factor in the difference between the control group and 30nm group (P0.05). The IL-12 factor in the control group and 30nm, the differences between 50nm groups (P0.05); can promote tumor and anti-tumor effect of inflammatory factor IL-10 in the control group, 30nm group at the same time, there are differences between groups 50nm (P0.05).3 The Xuanwei female lung cancer, peripheral blood: Xuanwei female lung cancer patients, tumor factors of female lung cancer patients in Xuanwei area and non healthy women in serum: factor IL-6 had significant differences between the normal group and Xuanwei group (P0.01), IL17 factor in normal group and Xuanwei group, there are differences between non Xuanwei group (P0.05). The antitumor factor: IL-4 factor differences between normal group and non Xuanwei group (P0.05), IL-13 in normal group and Xuanwei group, there were significant differences between non Xuanwei group (P0.01), IFN- gamma differences between normal group and Xuanwei group (P0.05); angiogenesis the formation and fibrosis factor: factor VEGF in normal group and Xuanwei group, there were significant differences between non Xuanwei group (P0.01), b-FGF factor differences between normal group and non Xuanwei group (P0.05), IL-12 factor in normal group and non Xuanwei group. There is a significant difference between Xuanwei group (P0.01); at the same time can promote tumor and anti-tumor effect of inflammatory factor IL-10 differences between normal group and Xuanwei group (P0.05), there is a significant difference between normal group and non Xuanwei group (P0.01). Conclusion: 1. The nano silicon oxide has a stimulating effect two on human bronchial epithelial cells, can promote the occurrence of severe bronchial epithelial dysplasia, may occur on.2 and lung cancer, by detecting the secretion of inflammatory state and fibrosis related cytokines to a certain extent, suggesting that changes in patients with.3, patients of Xuanwei female lung cancer peripheral blood cytokines are differentially expressed, may cause high incidence Yunnan Xuanwei female lung cancer.

【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R734.2

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相關(guān)期刊論文 前1條

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本文編號(hào)：1385768