应用BrdU-ELISA法检测化学物皮肤致敏性研究

Application of BrdU ELISA in the Detection of Chemical Skin Sensitization

目的研究应用BrdU-ELISA法检测化学物皮肤致敏性。方法将受试物2,4-二硝基氯苯(DNCB)、三氯乙烯(TCE)、三氯乙酸、抗菌冻胶对小鼠双耳背皮肤进行涂抹,25μl/耳,连续染毒3 d,第4天每只小鼠腹腔注射BrdU标记液(0.3ml/只)。第5天取小鼠耳部淋巴结称重,然后制成单细胞悬液,应用BrdU-ELISA法检测淋巴细胞增殖。同时应用豚鼠局部封闭涂皮法检测受试物的致敏性。结果DNCB、TCE各浓度组与AOO溶剂对照组BrdU标记值比较,差异均存在统计学意义(P<0.01),DNCB各浓度组与AOO溶剂对照组淋巴结重量比较,差异均存在统计学意义(P<0.01),而TCE各浓度组与AOO溶剂对照组淋巴结重量比较,差异无统计学意义(P>0.05)。抗菌凝胶组与三氯乙酸分别与空白对照组比较,其中抗菌凝胶组、1%、5%三氯乙酸BrdU标记值差异均无统计学意义,10%三氯乙酸差异则存在统计学意义(P<0.05)。结论BrdU-ELISA法具有检测化学物皮肤致敏性的能力,但其灵敏性比标准LLNA和传统豚鼠局部封闭涂皮法差,需要进一步优化实验方法。

[ Objective] To detect the chemical skin sensitization with BrdU ELISA. [ Methods] The chemicals, namely DNCB, TCE, TAC, antibiosis colloid were laid on the dorsum of the bilateral mice ears （25 μl/ear ） for three consecutive days; then the mice received an intraperitoneal injection of BrdU （ 0.3 ml/mouse ） on the fourth day ; on the fifth day, the lymph nodes of the mice were removed and weighted, then made into cell suspension, the lymph cell proliferation was measured with BrdU ELISA method; meanwhile the skin sensitization of DNCB, TCE, TAC and antibiosis colloid were tested using the guinea pig Buehler test. [ Resuits] Significant difference was found in the BrdU marker index between the DNCB and TCE groups of different concentration and the AOO solution control group （ P 〈 0.01 ） ; significant difference was also found in the lymph nodes weight between the DNCB groups of different concentrations and the AOO solution control group （ P 〈0.01 ）, but not between the TCE groups of different concentrations and the AOO solution control group （ P 〉 0.05 ） ; no significant difference was found in the BrdU marker index of the antibiosis colloid group and the 1% and 5% TAC groups, but significant difference was found in the 10% TAC group （ P 〈0.05）. [ Conclusion] BrdU ELISA has the ability to detect the chemical skin sensitization, but the sensitivity is no better than the guinea pig Buehler test and the Standard Local Lymph Node Assay.