本文選題：三氯乙烯　切入點(diǎn)：肝損傷　出處：《安徽醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的檢測(cè)并比較不同組豚鼠肝組織中TNF-α、IL-1、IL-6、IL-8表達(dá)水平,研究細(xì)胞因子在TCE致敏豚鼠肝損傷中的作用,探討三氯乙烯藥疹樣皮炎合并的肝損傷機(jī)制。 方法選用體重范圍為250 g左右白色雌性豚鼠45只,隨機(jī)分成空白對(duì)照組(5只)、溶劑對(duì)照組(5只)、TCE處理組(35只),依照豚鼠最大值實(shí)驗(yàn)(guinea pig maximization test,GPMT)方法建立動(dòng)物致敏模型,并在終末激發(fā)后依據(jù)致敏結(jié)果以及取材時(shí)點(diǎn)不同(分別于終末激發(fā)后24h、72h取材),將TCE處理組分為TCE致敏24h組、72h組和TCE未致敏24h組、72h組。觀察和記錄受試區(qū)的皮膚反應(yīng)情況并進(jìn)行拍照,按表一對(duì)皮膚反應(yīng)進(jìn)行評(píng)分。抽取豚鼠血液并處死豚鼠,在無(wú)菌條件下迅速取下豚鼠肝臟,部分做成蠟塊,部分做組織勻漿。于自動(dòng)生化分析儀上檢測(cè)肝功能,HE染色觀察病理學(xué)改變,免疫組織化學(xué)方法以及酶聯(lián)免疫吸附試驗(yàn)檢測(cè)豚鼠皮膚肝組織中TNF-α、IL-1、IL-6和IL-8的表達(dá)水平,用SPSS 13.0統(tǒng)計(jì)軟件包對(duì)實(shí)驗(yàn)結(jié)果進(jìn)行統(tǒng)計(jì)分析。 結(jié)果 1.在此次皮膚致敏反應(yīng)試驗(yàn)中,空白組和溶劑對(duì)照組均未見(jiàn)紅斑或水腫;TCE處理組的30只豚鼠中有19只出現(xiàn)紅斑,致敏率為63.3%。 2.肝功能TCE致敏組的ALT、AST水平較溶劑對(duì)照組升高,有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組ALT、AST水平顯著高于TCE致敏24h組以及TCE14d組,且有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組ALB水平下降,與溶劑對(duì)照組比較有統(tǒng)計(jì)學(xué)意義(p0.05)。 3.病理學(xué)觀察空白對(duì)照組、溶劑對(duì)照組豚鼠肝細(xì)胞排列整齊,胞質(zhì)均勻,胞核大而圓,核仁明顯,核膜清晰; TCE未致敏組能偶見(jiàn)肝細(xì)胞水腫;TCE致敏24h組中能夠見(jiàn)到少量肝細(xì)胞水腫;TCE致敏72h組中能夠見(jiàn)到較多肝細(xì)胞水腫,呈空泡樣改變,并有胞核破裂消失。 4.肝組織中TNF-α、IL-1、IL-6、IL-8水平檢測(cè) 4.1 TNF-α檢測(cè)結(jié)果免疫組化結(jié)果:TCE14d組、TCE致敏24h組、TCE致敏72h組TNF-α水平較溶劑對(duì)照組上升(p0.05);TCE致敏組肝組織中TNF-α水平分別高于各自未致敏組(p0.05);TCE致敏72h組肝組織中TNF-α水平要高于TCE致敏24h組以及TCE14d組(p0.05)。ELISA結(jié)果:TCE未致敏24h組、TCE致敏組24h組、TCE未致敏72h組、TCE致敏72h組肝組織中TNF-α水平高于溶劑對(duì)照組(p0.05);TCE致敏24h組、TCE致敏72h組肝組織中TNF-α水平分別高于各自未致敏組(p0.05);TCE致敏72h組、TCE致敏24h組肝組織中TNF-α水平均高于TCE14d組水平p0.05)。 4.2 IL-1檢測(cè)結(jié)果免疫組化結(jié)果:與溶劑對(duì)照組比較,TCE致敏24h組、TCE致敏72h組以及TCE未致敏24h組肝組織中IL-1β水平升高(p0.05);TCE致敏組24h、TCE致敏72h組肝組織中IL-1β水平分別高于各自未至敏組(p0.05);TCEh致敏24組、TCE致敏72h組肝組織中IL-1β水平均高于T14d組(p0.05);而且,TCE致敏72h組肝組織中IL-1β水平要高于TCE致敏24h組(p0.05)。ELISA結(jié)果:TCE處理的各組肝組織中IL-1水平較溶劑對(duì)照組都上升,除TCE未致敏72h組外,其它組有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組肝組織中IL-1水平高于TCE未致敏72h組(p0.05);TCE致敏72h組肝組織中IL-1水平高于TCE致敏24h組、TCE14d組(p0.05)。 4.3 IL-6檢測(cè)結(jié)果免疫組化結(jié)果:TCE致敏24h組、TCE致敏72h組肝組織中IL-6水平較溶劑對(duì)照組升高(p0.05);TCE致敏24h組、TCE致敏72h組肝組織中IL-6水平分別高于各自未至敏組(p0.05);TCE致敏24h組、TCE致敏72h組肝組織中IL-6水平均高于T14d組(p0.05)。ELISA結(jié)果:TCE致敏組24h組、TCE未致敏72h組、TCE致敏72h組較溶劑對(duì)照組升高,差異且有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組肝組織中IL-6水平高于TCE未致敏72h組,差異有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組、TCE致敏24h組肝組織中IL-6水平均高于TCE14d組水平,其中TCE致敏72h組有統(tǒng)計(jì)學(xué)意義(p0.05)。 4.4 IL-8檢測(cè)結(jié)果免疫組化結(jié)果:與溶劑對(duì)照比較,TCE致敏24h組、TCE致敏72h組肝組織中IL-8水平升高(p0.05);TCE致敏24h組、TCE致敏72h組肝組織中IL-8水平分別高于各自未致敏組(p0.05);TCE致敏24h組、TCE致敏72h組肝組織中IL-8水平均高于T14d組(p0.05)。ELISA結(jié)果:TCE處理的各組肝組織中IL-8水平都上升,且有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組肝組織中IL-8水平高于TCE未致敏72h組,差異有統(tǒng)計(jì)學(xué)意義(p0.05);TCE致敏72h組、TCE致敏24h組肝組織中IL-8水平均高于TCE14d組水平,其中TCE致敏72h組有統(tǒng)計(jì)學(xué)意義(p0.05)。 5.肝組織中細(xì)胞因子以及細(xì)胞因子與肝功能間相關(guān)性分析 TNF-α與IL-1、IL-6、IL-8之間存在顯著正相關(guān),即隨著肝組織中TNF-α水平的上升,肝組織中IL-1、IL-6、IL-8的表達(dá)也增加;IL-1與IL-6、IL-8之間也存在顯著正相關(guān),但I(xiàn)L-6與IL-8相關(guān)性無(wú)統(tǒng)計(jì)學(xué)意義。ALT、AST與TNF-α、IL-6呈正相關(guān),且有統(tǒng)計(jì)學(xué)意義(p0.05);除與TNF-α呈負(fù)相關(guān)有統(tǒng)計(jì)學(xué)意義(p0.05),ALB與IL-1、IL-6、IL-8相關(guān)性無(wú)統(tǒng)計(jì)學(xué)意義。 結(jié)論:本研究證實(shí)TCE對(duì)豚鼠具有致敏作用;能夠引起致敏豚鼠不同程度的肝損傷;TCE致敏豚鼠肝組織內(nèi)TNF-α、IL-1、IL-6、IL-8水平較其它組高,且ALT、AST與TNF-α、IL-6呈正相關(guān),提示TNF-α、IL-1、IL-6、IL-8在三氯乙烯致敏豚鼠肝損傷的過(guò)程中發(fā)揮重要作用。
[Abstract]:Objective to detect and compare the expression levels of TNF-, IL-1, IL-6 and IL-8 in different groups of guinea pig liver tissues, and investigate the role of cytokines in the liver injury induced by TCE sensitized guinea pigs, and to explore the mechanism of liver injury associated with trichloroethylene induced drug eruption dermatitis.
Methods the weight range is about 250 g 45 white female guinea pigs were randomly divided into control group (5 rats), solvent control group (5 rats), TCE group (35), in accordance with the maximum value of experimental guinea pigs (guinea pig maximization test, GPMT) method to establish animal model of sensitization, and in the end at the end of excitation according to sensitization results and different sampling point (respectively excited at the terminal of 24h, 72h, were) TCE treatment group was divided into TCE sensitized 24h group, 72h group and TCE sensitized 24h group and 72h group. Observe and record the skin reaction test area and take photos, according to a table of skin reaction score. Extraction of guinea pig blood and guinea pigs were sacrificed, quickly remove the guinea pig liver under aseptic conditions, some wax blocks, part of homogenate. In automatic biochemical analyzer to detect liver function, pathological changes were observed with HE staining, immunohistochemistry and enzyme-linked immunosorbent assay The expression level of TNF- alpha, IL-1, IL-6 and IL-8 in the skin liver tissues of guinea pigs was detected, and the experimental results were statistically analyzed with the statistical package of SPSS 13.
Result
1. in the skin sensitization test, there were no erythema or edema in the blank group and the solvent control group. 19 of the 30 guinea pigs in the TCE treated group had erythema, and the sensitization rate was 63.3%..
2. liver function TCE ALT sensitized group, AST level compared with the solvent control group increased, with statistical significance (P0.05); TCE sensitized 72h group ALT, AST was significantly higher than that of TCE sensitized 24h group and TCE14d group, and there was statistical significance (P0.05); TCE sensitized 72h group ALB decreased, and the solvent the control group was statistically significant (P0.05).
Observation of the blank control group, solvent control group 3. pathology of guinea pig liver cells arranged in neat, uniform cytoplasm, large round nucleus, obvious nucleolus, nuclear membrane was clear; TCE sensitized group can even see liver cell edema; TCE sensitized 24h group to be able to see a small amount of liver cell edema; TCE sensitized 72h group to see more liver cell edema, vacuolar changes, and nucleus broken away.
Detection of TNF- alpha, IL-1, IL-6, IL-8 in 4. liver tissues
4.1 TNF- alpha test results the results of immunohistochemistry: TCE14d group, TCE sensitized 24h group and TCE sensitized 72h group TNF- levels than the solvent control group increased (P0.05); TCE TNF- induced by alpha level of liver tissue were higher than their respective sensitive sensitized group (P0.05); TCE TNF- induced by alpha level of liver sensitive tissue in the 72h group than the TCE sensitized 24h group and TCE14d group (P0.05.ELISA) results: TCE sensitized 24h group and TCE sensitized group 24h group TCE sensitized 72h group, TCE induced TNF- levels of liver tissue sensitivity to 72h was significantly higher than that of the solvent control group (P0.05); TCE sensitization 24h group, TCE induced TNF- levels of liver tissue in 72h group were higher than that of their sensitive non sensitized group (P0.05); TCE sensitized 72h group, TCE induced TNF- levels of liver tissue sensitivity was higher in group 24h than in group TCE14d P0.05).
4.2 IL-1媯，

本文編號(hào)：1600932