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目的建立小鼠局部淋巴结检测结合耳肿胀程度评估的方法鉴别光敏物。方法受试物加UVA照射(剂量10J/cm2)连续3d在BALB/c小鼠耳背部进行激发,最后一次照射结束后24h切取引流的局部淋巴结,用淋巴细胞计数和MTT比色法测定淋巴细胞增殖率反映局部淋巴结的增殖情况;同时测量小鼠耳肿度以反映皮肤的刺激反应;用双抗体夹心ELISA法进一步测定局部淋巴结细胞72h培养上清液中干扰素-γ(IFN-γ),白介素-2(IL-2)以及白介素-4(IL-4)的含量。结果强光变应原四氯水杨酰苯胺引起局部淋巴结的增殖反应而不伴耳肿胀反应;弱光变应原6甲基香豆素既不引起局部淋巴结的增殖反应又不出现耳肿胀反应;光毒物质8-甲氧补骨脂素同时引起局部淋巴结增殖和耳肿胀反应。用四氯水杨酰苯胺预先致敏后再进行激发,局部淋巴结中IL-4的分泌量明显升高(P<0.05)。结论小鼠局部淋巴结检测结合耳肿胀程度评价的方法可较快鉴定较强的接触性光变应原和光毒物质,但对识别较弱的光变应原敏感性不足;预先致敏后激发阶段局部淋巴结中IL-4的分泌量明显升高是光变应原区别于光毒物质的突出特点。
Objective To establish a method to identify photosensitizers by detecting the degree of ear swelling in mice with local lymph nodes. Methods The subjects were irradiated with UVA (dose of 10 J / cm2) for 3 consecutive days in BALB / c mice. The draining lymph nodes were excised 24 h after the last irradiation. Lymphocyte count and MTT colorimetric assay Cell proliferation rate was measured to reflect the proliferation of local lymph nodes. Ear swelling of mice was also measured to reflect skin irritation. Interferon-γ (IFN-γ) expression in local lymph node cells was detected by double antibody sandwich ELISA. , Interleukin-2 (IL-2) and interleukin-4 (IL-4) levels. Results The light allergen tetracycline salicylanilide induced the proliferative response of the local lymph nodes without ear swelling. The weakly light allergen 6 methylcoumarin did not cause the proliferation reaction of the local lymph nodes without ear swelling reaction ; Phototoxic substances 8-methoxypsoralen also cause local lymph node proliferation and ear swelling reaction. After sensitization with tetrachlorosalicylanilide, the secretion of IL-4 in local lymph nodes was significantly increased (P <0.05). Conclusion The method of local lymph node detection combined with the evaluation of ear swelling can identify strong contact photoproteins and phototoxins more quickly but is less sensitive to the weaker recognition photoreagens. The pre-sensitization post-sensitization phase Local lymph node IL-4 secretion was significantly increased light allergen different from the prominent features of light toxin.