本文選題：二氧化硅 + 矽肺��； 參考：《重慶醫(yī)科大學(xué)》2013年碩士論文

【摘要】：矽肺是因長期大量吸入游離二氧化硅(SiO2)粉塵所引起的以肺組織彌漫性纖維化為主要特征的全身性疾病。根據(jù)2011年4月18日衛(wèi)生部向社會公布的《2010年職業(yè)病防治工作情況和2011年重點(diǎn)工作》，2010年新發(fā)職業(yè)病27240例，其中塵肺病23812例。目前塵肺病仍是我國最嚴(yán)重的職業(yè)病，2010年報(bào)告的塵肺病例數(shù)占職業(yè)病報(bào)告總例數(shù)的87.42%。有專家調(diào)查發(fā)現(xiàn)，能夠做職業(yè)病診斷并在疾控中心登記的，僅為塵肺病患者的10%-20%，因此估計(jì)塵肺病實(shí)際發(fā)生的病例數(shù)不少于100萬人。因此，進(jìn)一步探討矽肺的發(fā)病機(jī)制和尋找新型的治療藥物對矽肺的預(yù)防、治療乃至最終消除矽肺具有重要的意義。目前的研究表明，T淋巴細(xì)胞由樹突狀細(xì)胞和巨噬細(xì)胞等抗原遞呈細(xì)胞處理修飾二氧化硅后激活。雖然Th1/Th2極化在矽肺的發(fā)病機(jī)制中仍不完全明確，但研究表明，Th1型細(xì)胞因子在矽肺的炎癥階段起主要作用，而Th2型細(xì)胞因子在矽肺的纖維化階段起主要作用。IFN-γ（γ-干擾素，Interferon-γ）具有促進(jìn)初始T淋巴細(xì)胞向Th1型細(xì)胞分化以及活化巨噬細(xì)胞和NK細(xì)胞的作用，是典型的Th1型細(xì)胞因子。Th1型細(xì)胞的分化成熟需要IFN-γ依賴的信號轉(zhuǎn)導(dǎo)子和轉(zhuǎn)錄激活子1（signaltransducers and activators of transcription1,STAT1）STAT1和STAT4的磷酸化信號級聯(lián)反應(yīng)。實(shí)驗(yàn)研究證實(shí)，富含重復(fù)“TTAGGG”基序的抑制性寡核苷酸（suppressive oligodeoxynucleotides, Sup ODN）具有下調(diào)Th1型細(xì)胞因子和前炎癥介質(zhì)產(chǎn)生的作用。因此，我們可以將這類ODN用于矽肺的研究中，觀察其對矽肺早期炎癥的影響，以及炎癥細(xì)胞因子的產(chǎn)生影響。 研究目的 觀察Sup ODN對染矽塵小鼠早期炎癥的影響，探討Sup ODN作用機(jī)制并評價(jià)其應(yīng)用價(jià)值。研究方法 雌性Balb/c小鼠60只隨機(jī)分4組，分別為空白對照組、矽塵組、Sup ODN組、對照寡核苷酸（control oligodeoxynucleotides, Con ODN)組。除空白對照組氣管注入0.1mL生理鹽水外，其余各組采用氣管暴露法一次性注入0.1mL SiO2懸液（5g/L），Sup ODN組和Con ODN組在造模前3h分別腹腔注射Sup ODN、Con ODN各0.3mL(1mg/mL)。以建模術(shù)時(shí)為0d記，模型建立7d后處死動物，，采用瑞氏染色作血清白細(xì)胞分類計(jì)數(shù)；ELISA法檢測血清IFN-γ和TNF-α水平；免疫組織化學(xué)法檢測肺組織中IFN-γ和pSTAT4的表達(dá),并作定量分析；HE染色觀察肺組織病理變化；天狼猩紅染色觀察肺組織Ⅰ、Ⅲ型膠原表達(dá)。 研究結(jié)果 1.HE染色證實(shí)氣管暴露注入二氧化硅粉塵懸液成功建立了小鼠矽肺模型。 2.Sup ODN組與矽塵組、Con ODN組相比，血清中IFN-γ表達(dá)明顯降低（P0.01）；矽塵組小鼠肺組織間隔水腫并增厚，Sup ODN組小鼠肺組織炎癥較矽塵組減輕；與空白對照組比較，矽塵組小鼠肺組織中IFN-γ表達(dá)增加(P0.05)，與矽塵組比較，Sup ODN組小鼠肺組織內(nèi)IFN-γ表達(dá)減少(P0.05);與空白對照組比較，矽塵組小鼠肺組織中pSTAT4表達(dá)顯著增加(P0.01)，與矽塵組比較，Sup ODN組小鼠肺組織內(nèi)pSTAT4表達(dá)顯著減少(P0.01)。 研究結(jié)論 1.矽塵可誘導(dǎo)染矽塵小鼠血清中IFN-γ和TNF-α表達(dá)增加，SupODN可減少染矽塵小鼠IFN-γ的產(chǎn)生，而對TNF-α無影響。 2.Sup ODN可能通過抑制IFN-γ和pSTAT4的表達(dá)，抑制小鼠Th1型免疫應(yīng)答，最終實(shí)現(xiàn)其對染矽塵小鼠肺炎癥的保護(hù)作用。綜上所述，本研究結(jié)果初步表明Sup ODN對染矽塵小鼠早期炎癥有一定的阻礙作用，為矽肺炎癥的防治及輔助治療，開拓了新的途徑。
[Abstract]:Silicosis is a systemic disease characterized by diffuse fibrosis of free silica (SiO2) for a long time. According to the status of occupational disease prevention and Prevention published in April 18, 2011 by the Ministry of health and the key work in 2011, 27240 new occupational diseases were found in 2010, including 23812 cases of pneumoconiosis. At present, pneumoconiosis is still the most serious occupational disease in China. The number of pneumoconiosis cases reported in 2010 accounted for 87.42%. of the total number of occupational diseases reported by experts. It was found that the number of cases of pneumoconiosis diagnosed and registered at the CDC was only 10%-20% of pneumoconiosis patients, so the number of cases of pneumoconiosis was estimated to be not less than 1 million. One step to explore the pathogenesis of silicosis and the search for new therapeutic drugs is of great significance for the prevention of silicosis, treatment and even eventually eliminating silicosis. The present study shows that T lymphocytes are activated by dendritic cells and macrophages and other antigen presenting cells to treat the modified silica. Although Th1/Th2 polarization is in the pathogenesis of silicosis The system is still not completely clear, but the study shows that type Th1 cytokines play a major role in the inflammatory stage of silicosis, while Th2 type cytokines play a major role in the fibrosis stage of silicosis..IFN- gamma (interferon gamma, Interferon- gamma) can promote the differentiation of initial T lymphocytes to Th1 cells and the activation of macrophages and NK cells. The differentiation of typical Th1 type cytokine.Th1 type cells requires IFN- gamma dependent signal transducers and transcriptional activator 1 (signaltransducers and activators of transcription1, STAT1) STAT1 and STAT4 phosphorylation signal cascade reaction. Ligodeoxynucleotides, Sup ODN) has the role of down-regulation of Th1 type cytokines and proinflammatory mediators. Therefore, we can use this kind of ODN in silicosis research to observe its effect on early silicosis and the effect of inflammatory cytokines.
research objective
To observe the effect of Sup ODN on the early inflammation of mice exposed to silica dust, to explore the mechanism of Sup ODN and evaluate its application value.
60 female Balb/c mice were randomly divided into 4 groups: blank control group, silica dust group, Sup ODN group and control oligonucleotide (control oligodeoxynucleotides, Con ODN) group. Except the blank control group, the trachea was injected into the 0.1mL physiological saline, the rest of the other groups were injected into the 0.1mL SiO2 suspension (5g/L) by tracheal exposure method. The anterior 3H was intraperitoneally injected with Sup ODN and Con ODN 0.3mL (1mg/mL). The animal was killed after modeling, and the animal was killed after the model was established. The serum white blood cell count was counted by Rayleigh staining. The ELISA method was used to detect the level of IFN- gamma and TNF- alpha in serum; the immunohistochemical method was used to detect the expression of gamma ray in lung tissue and quantitative analysis. The pathological changes of lung tissue were observed. The expression of type I and III collagen in lung tissue was observed by Sirius red staining.
Research results
1.HE staining confirmed that the silicosis model was successfully established by injecting silica dust into the trachea.
In group 2.Sup ODN, the expression of IFN- gamma in serum was significantly lower than that in Con ODN group (P0.01), and the pulmonary tissue was thicker and thicker in the silico dust group, and the lung tissue inflammation in the Sup ODN group was less than that in the silica dust group. Compared with the blank control group, the expression of IFN- gamma in the lung tissue of the silicodust mice increased (P0.05), and the Sup ODN group lung was compared with that of the silica dust group. The expression of IFN- gamma in the tissue was decreased (P0.05), and the expression of pSTAT4 in the lung tissue of the silica dust group was significantly increased (P0.01) compared with the blank control group (P0.01). The expression of pSTAT4 in the lung tissue of the Sup ODN group was significantly reduced (P0.01).
research conclusion
1. silica dust can induce the expression of IFN- gamma and TNF- alpha in serum of mice exposed to silica dust. SupODN can reduce the production of IFN- gamma in mice infected with silica dust, but has no effect on TNF- alpha.
2.Sup ODN may inhibit the Th1 type immune response in mice by inhibiting the expression of IFN- gamma and pSTAT4 and finally realizing its protective effect on pneumonia in mice infected with silica dust. To sum up, the results of this study preliminarily indicate that Sup ODN has a certain hindrance effect on early inflammation in mice infected with silicosis, and has opened a new way for the prevention and treatment of silicosis inflammation and adjuvant therapy. Way.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R135.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 高衍新;王瑞;;矽塵致肺纖維化機(jī)制及細(xì)胞因子在矽肺纖維化中的作用[J];中國工業(yè)醫(yī)學(xué)雜志;2008年01期

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