本文選題：三氯乙烯　切入點(diǎn)：豚鼠　出處：《安徽醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 1研究背景 三氯乙烯(Trichloroethylene, TCE)是一種常見的鹵代烴類工業(yè)有機(jī)溶劑,它是脂溶性易揮發(fā)的物質(zhì),無(wú)色、不易燃燒液體,有類似氯仿的氣味,可用做金屬去脂劑、干洗劑、溶劑、萃取劑等,每年有5-6萬(wàn)噸的TCE使用量,在五金、電子、玩具、印刷等行業(yè)應(yīng)用廣泛。在過(guò)去的研究中對(duì)其致機(jī)體的損傷作用主要集中于對(duì)腎臟、中樞和末梢神經(jīng)系統(tǒng)、心臟、生殖系統(tǒng)等器官損傷研究。近年來(lái)的研究發(fā)現(xiàn)其還可以導(dǎo)致嚴(yán)重的皮膚損傷,許多學(xué)者認(rèn)為TCE造成的皮膚損傷屬于T淋巴細(xì)胞介導(dǎo)的Ⅳ型遲發(fā)性變態(tài)反應(yīng),然而臨床上僅用T淋巴細(xì)胞介導(dǎo)的Ⅳ型遲發(fā)性變態(tài)反應(yīng)尚不能完全解釋其皮膚損傷機(jī)制。同時(shí)近年來(lái)還發(fā)現(xiàn)不少死亡病例的病因與嚴(yán)重的肝損傷引起的多臟器功能衰竭有關(guān),因此由TCE致敏引起的肝損傷引起了人們的高度重視,而根據(jù)對(duì)TCE藥疹樣皮炎的病例的資料調(diào)查后發(fā)現(xiàn),TCE藥疹樣皮炎的病人常常伴有嚴(yán)重肝損傷,而肝損傷是引起多臟器衰竭導(dǎo)致死亡的主要原因。工作場(chǎng)所接觸到外源化合物而導(dǎo)致的皮膚炎性疾患統(tǒng)稱為職業(yè)性接觸性皮炎(Occupational contact dermatitis, OCD),目前根據(jù)發(fā)病機(jī)理不同通常將接觸性皮炎分為刺激性接觸性皮炎(Irritant contact dermatitis, ICD)和過(guò)敏性接觸性皮炎(Allergic contact dermatitis, ACD),由于僅用T淋巴細(xì)胞介導(dǎo)的Ⅳ型遲發(fā)性變態(tài)反應(yīng)可能尚不能完全解釋TCE所致皮膚損傷的機(jī)制,表明TCE所致藥疹樣皮膚損傷可能并非只是T淋巴細(xì)胞介導(dǎo)的Ⅳ型遲發(fā)性變態(tài)反應(yīng)。 2目的 研究TCE對(duì)豚鼠的皮膚致敏作用及其作用機(jī)制,通過(guò)檢測(cè)豚鼠肝功能相關(guān)指標(biāo)研究TCE致敏導(dǎo)致對(duì)肝臟的損傷作用。通過(guò)檢測(cè)氧化應(yīng)激相關(guān)指標(biāo),探討TCE致敏豚鼠肝損傷的可能機(jī)制。 3方法 選用體重250-300g/只,白色雌性豚鼠,經(jīng)適應(yīng)性飼養(yǎng)一星期后,隨機(jī)分成空白對(duì)照組、溶劑對(duì)照組(橄欖油)、TCE組和DNCB陽(yáng)性對(duì)照組。用電動(dòng)剃刀去除背部毛,約4cm-6cm,24h后取皮膚完整無(wú)損傷得動(dòng)物隨機(jī)分組供試。豚鼠背部從頭向尾設(shè)L1;R1;L2;R2;L3;R3共3對(duì)6個(gè)點(diǎn),各點(diǎn)間距1.5cm,各對(duì)間距3cm。L1、R1兩點(diǎn)注射0.1ml弗氏完全佐劑(FCA);L2、R2兩點(diǎn)注射0.1ml受試物;L3、R3兩點(diǎn)注射0.1mlFCA與受試物的等量混合物。第1d用5% TCE(TCE:橄欖油=5:95)和0.25% DNCB(DNCB:丙酮:橄欖油=1:3:397)皮內(nèi)注射進(jìn)行初次致敏,對(duì)照組注射L1;R1兩點(diǎn)FCA,L2、R2兩點(diǎn)為未稀釋的賦形劑,L3、R3兩點(diǎn)則用1:1(V/V)FCA/水或生理鹽水配制的濃度為50%(w/v)的賦形劑。第7d在注射部位涂抹10%十二烷基硫酸鈉,于第8d用40% TCE涂皮和0.50% DNCB涂皮進(jìn)行第二次致敏,48h后用生理鹽水清洗,在第15d用20% TCE涂皮和0.25% DNCB涂皮進(jìn)行激發(fā),24h后用生理鹽水清洗。依據(jù)《化學(xué)品毒性鑒定技術(shù)規(guī)范》(衛(wèi)監(jiān)督發(fā)【2005】272號(hào))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2.1對(duì)皮膚反應(yīng)進(jìn)行評(píng)分。當(dāng)受試樣品組動(dòng)物皮膚反應(yīng)積分≥1時(shí),應(yīng)判為皮膚致敏反應(yīng)陽(yáng)性,按表2.2對(duì)受試樣品進(jìn)行致敏強(qiáng)度分級(jí)。 將豚鼠血液采出后,制備血清,采用試劑盒測(cè)定丙二醛(Malondialdehyde, MDA ),超氧化物歧化酶(Superoxide dismutase, SOD)含量和活性。在自動(dòng)生化儀上測(cè)定與肝功能相關(guān)的指標(biāo),包括ALT、AST、ALB等指標(biāo),用SPSS10.0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。 4結(jié)果 1、在皮膚致敏反應(yīng)試驗(yàn)中,空白組和橄欖油對(duì)照組均未見紅斑或水腫, DNCB陽(yáng)性對(duì)照組7只動(dòng)物均出現(xiàn)紅斑以及水腫反應(yīng),致敏率為100%;在TCE試驗(yàn)組的14只動(dòng)物中有9只出現(xiàn)紅斑,致敏率為64.29%。 2、在對(duì)豚鼠肝臟進(jìn)行肝功能檢查發(fā)現(xiàn),致敏豚鼠肝臟ALT、AST指標(biāo)與空白組和橄欖油對(duì)照組均差異有顯著性;在同批激發(fā)的豚鼠中,致敏陽(yáng)性組的ALT、AST指標(biāo)比致敏陰性組的豚鼠明顯升高,經(jīng)檢驗(yàn)差異有顯著性;在對(duì)DNCB陽(yáng)性對(duì)照組的豚鼠的肝臟ALT、AST指標(biāo)檢測(cè)后發(fā)現(xiàn),陽(yáng)性對(duì)照組和空白組以及橄欖油對(duì)照組相比,ALT、AST指標(biāo)差異無(wú)顯著性。 3、在對(duì)豚鼠血清中的脂質(zhì)過(guò)氧化水平檢測(cè)后發(fā)現(xiàn),致敏豚鼠血清的MDA的水平和對(duì)照組相比呈降低的趨勢(shì),經(jīng)統(tǒng)計(jì)學(xué)檢驗(yàn)差異有顯著性;在同批激發(fā)的豚鼠中,致敏陽(yáng)性組和致敏陰性組豚鼠的MDA水平相比,致敏陽(yáng)性豚鼠的MDA明顯降低;豚鼠血清的SOD水平檢測(cè)后發(fā)現(xiàn),致敏陽(yáng)性豚鼠和對(duì)照組相比呈升高趨勢(shì),經(jīng)統(tǒng)計(jì)學(xué)檢驗(yàn)差異有顯著性;TCE致敏陽(yáng)性組和TCE致敏陰性組豚鼠的SOD水平相比,陽(yáng)性組SOD水平明顯升高。 5結(jié)論 1、TCE在一定劑量下對(duì)豚鼠具有致敏作用,屬于強(qiáng)致敏物。 2、經(jīng)TCE致敏豚鼠的肝臟有一定的損傷,而不僅僅是皮膚的損傷,且這種損傷是與TCE的作用有關(guān)的。TCE導(dǎo)致致敏豚鼠體內(nèi)脂質(zhì)過(guò)氧化反應(yīng)水平發(fā)生改變,抗氧化酶SOD的活性升高,而MDA水平降低。其損傷機(jī)制可能由于脂質(zhì)過(guò)氧化反應(yīng)造成肝損害,但自由基引起損傷的具體機(jī)制還需進(jìn)一步研究。
[Abstract]:1 background of research
Trichloroethylene (Trichloroethylene, TCE) is a common halogenated hydrocarbon, industrial organic solvent, it is fat soluble volatile substances, colorless, nonflammable liquid, chloroform like odor, can be used as a metal degreasing agent, cleaning agent, solvent, extraction agent, 5-6 tons of TCE usage. Every year, in hardware, electronics, toys, printing and other industries widely used. Damage caused by the body in the past research mainly focused on the kidney, central and peripheral nervous system, heart, and other organs damage on reproductive system in recent years. The research found that it can also cause serious skin damage, many scholars believe that the type IV skin injury caused by TCE belongs to the T lymphocyte mediated delayed allergic reaction, but only by clinical type IV T lymphocyte mediated delayed allergic reaction can not fully explain the skin damage mechanism. At the same time in recent years Find the cause of many deaths and serious liver injury multiple organ failure related to liver injury, so sensitized by TCE has aroused people's attention, and according to the survey data of TCE cases of dermatitis medicamentosa like dermatitis medicamentosa like TCE found that patients often accompanied by severe liver injury, liver injury is the leading cause of death due to multiple organ failure. Inflammatory disorders of the workplace exposure to exogenous compounds caused by the skin called occupation contact dermatitis (Occupational contact, dermatitis, OCD), according to the different pathogenesis of contact dermatitis usually consists of irritant contact dermatitis (Irritant contact dermatitis, ICD) and allergic contact dermatitis (Allergic contact, dermatitis, ACD), due to the delayed allergic reaction may not explain completely with only TC type IV mediated by T lymphocytes The mechanism of skin injury caused by E indicates that the skin damage caused by TCE induced rash like skin may not be only the type IV delayed allergic reaction mediated by T lymphocyte.
2 purposes
Objective to study the sensitization effect of TCE on guinea pig skin and its action mechanism, and to study the injury effect of TCE sensitization on liver by detecting relevant indexes of guinea pig liver function. To explore the possible mechanism of TCE sensitized guinea pig liver injury by detecting oxidative stress related indicators.
3 method
閫夌敤浣撻噸250-300g/鍙，

本文編號(hào)：1727254