三氯乙烯致豚鼠皮肤和肝脏免疫性损伤

发布时间：2018-04-09 16:37

  本文选题：三氯乙烯　切入点：豚鼠　出处：《安徽医科大学》2008年硕士论文

【摘要】： 1研究背景 三氯乙烯(Trichloroethylene, TCE)是一种常见的卤代烃类工业有机溶剂,它是脂溶性易挥发的物质,无色、不易燃烧液体,有类似氯仿的气味,可用做金属去脂剂、干洗剂、溶剂、萃取剂等,每年有5-6万吨的TCE使用量,在五金、电子、玩具、印刷等行业应用广泛。在过去的研究中对其致机体的损伤作用主要集中于对肾脏、中枢和末梢神经系统、心脏、生殖系统等器官损伤研究。近年来的研究发现其还可以导致严重的皮肤损伤,许多学者认为TCE造成的皮肤损伤属于T淋巴细胞介导的Ⅳ型迟发性变态反应,然而临床上仅用T淋巴细胞介导的Ⅳ型迟发性变态反应尚不能完全解释其皮肤损伤机制。同时近年来还发现不少死亡病例的病因与严重的肝损伤引起的多脏器功能衰竭有关,因此由TCE致敏引起的肝损伤引起了人们的高度重视,而根据对TCE药疹样皮炎的病例的资料调查后发现,TCE药疹样皮炎的病人常常伴有严重肝损伤,而肝损伤是引起多脏器衰竭导致死亡的主要原因。工作场所接触到外源化合物而导致的皮肤炎性疾患统称为职业性接触性皮炎(Occupational contact dermatitis, OCD),目前根据发病机理不同通常将接触性皮炎分为刺激性接触性皮炎(Irritant contact dermatitis, ICD)和过敏性接触性皮炎(Allergic contact dermatitis, ACD),由于仅用T淋巴细胞介导的Ⅳ型迟发性变态反应可能尚不能完全解释TCE所致皮肤损伤的机制,表明TCE所致药疹样皮肤损伤可能并非只是T淋巴细胞介导的Ⅳ型迟发性变态反应。 2目的 研究TCE对豚鼠的皮肤致敏作用及其作用机制,通过检测豚鼠肝功能相关指标研究TCE致敏导致对肝脏的损伤作用。通过检测氧化应激相关指标,探讨TCE致敏豚鼠肝损伤的可能机制。 3方法 选用体重250-300g/只,白色雌性豚鼠,经适应性饲养一星期后,随机分成空白对照组、溶剂对照组(橄榄油)、TCE组和DNCB阳性对照组。用电动剃刀去除背部毛,约4cm-6cm,24h后取皮肤完整无损伤得动物随机分组供试。豚鼠背部从头向尾设L1;R1;L2;R2;L3;R3共3对6个点,各点间距1.5cm,各对间距3cm。L1、R1两点注射0.1ml弗氏完全佐剂(FCA);L2、R2两点注射0.1ml受试物;L3、R3两点注射0.1mlFCA与受试物的等量混合物。第1d用5% TCE(TCE:橄榄油=5:95)和0.25% DNCB(DNCB:丙酮:橄榄油=1:3:397)皮内注射进行初次致敏,对照组注射L1;R1两点FCA,L2、R2两点为未稀释的赋形剂,L3、R3两点则用1:1(V/V)FCA/水或生理盐水配制的浓度为50%(w/v)的赋形剂。第7d在注射部位涂抹10%十二烷基硫酸钠,于第8d用40% TCE涂皮和0.50% DNCB涂皮进行第二次致敏,48h后用生理盐水清洗,在第15d用20% TCE涂皮和0.25% DNCB涂皮进行激发,24h后用生理盐水清洗。依据《化学品毒性鉴定技术规范》(卫监督发【2005】272号)观察和记录受试区的皮肤反应情况并进行拍照。按表2.1对皮肤反应进行评分。当受试样品组动物皮肤反应积分≥1时,应判为皮肤致敏反应阳性,按表2.2对受试样品进行致敏强度分级。 将豚鼠血液采出后,制备血清,采用试剂盒测定丙二醛(Malondialdehyde, MDA ),超氧化物歧化酶(Superoxide dismutase, SOD)含量和活性。在自动生化仪上测定与肝功能相关的指标,包括ALT、AST、ALB等指标,用SPSS10.0统计软件包对实验结果进行统计分析。 4结果 1、在皮肤致敏反应试验中,空白组和橄榄油对照组均未见红斑或水肿, DNCB阳性对照组7只动物均出现红斑以及水肿反应,致敏率为100%;在TCE试验组的14只动物中有9只出现红斑,致敏率为64.29%。 2、在对豚鼠肝脏进行肝功能检查发现,致敏豚鼠肝脏ALT、AST指标与空白组和橄榄油对照组均差异有显著性;在同批激发的豚鼠中,致敏阳性组的ALT、AST指标比致敏阴性组的豚鼠明显升高,经检验差异有显著性;在对DNCB阳性对照组的豚鼠的肝脏ALT、AST指标检测后发现,阳性对照组和空白组以及橄榄油对照组相比,ALT、AST指标差异无显著性。 3、在对豚鼠血清中的脂质过氧化水平检测后发现,致敏豚鼠血清的MDA的水平和对照组相比呈降低的趋势,经统计学检验差异有显著性;在同批激发的豚鼠中,致敏阳性组和致敏阴性组豚鼠的MDA水平相比,致敏阳性豚鼠的MDA明显降低;豚鼠血清的SOD水平检测后发现,致敏阳性豚鼠和对照组相比呈升高趋势,经统计学检验差异有显著性;TCE致敏阳性组和TCE致敏阴性组豚鼠的SOD水平相比,阳性组SOD水平明显升高。 5结论 1、TCE在一定剂量下对豚鼠具有致敏作用,属于强致敏物。 2、经TCE致敏豚鼠的肝脏有一定的损伤,而不仅仅是皮肤的损伤,且这种损伤是与TCE的作用有关的。TCE导致致敏豚鼠体内脂质过氧化反应水平发生改变,抗氧化酶SOD的活性升高,而MDA水平降低。其损伤机制可能由于脂质过氧化反应造成肝损害,但自由基引起损伤的具体机制还需进一步研究。
[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，

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