[广谱半胱天冬酶抑制剂Z-VAD-FMK对过敏性接触性皮炎的抑制作用：小鼠实验]

[Attenuation of allergic contact dermatitis by Z-VAD-FMK, a broad caspase inhibitor: experiment with mice].

作者信息

Li Yuan-Yuan, Yan Chun-Lin, Xu Wei

机构信息

Department of Dermatology, Huashan Hospital, Fudan University, Shanghai 200040, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2008 Dec 2;88(44):3153-6.

PMID:19159602

Abstract

OBJECTIVE

To investigate the effects of N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK), a broad caspase inhibitor, on allergic contact dermatitis (ACD).

METHODS

A Balb/c mouse was killed and its back skin was peeled, put into a Franz diffusion cell, and smeared with Z-VAD-FMK. High performance liquid chromatography was used to examine the permeation rate. Dinitro-fluorobenzene (DNFB) was applied on the depilated back of mice as inducer and then on the back-side of right ear as provocative agent 6 days later so as to establish mouse ACD models. Two hours after the provocation 24 mouse models were divided into 4 equal groups to undergo the application of Z-VAD-FMK at the concentration of 0.1, 0.5, 2.5, and 5 mmol/L on the back-side of right ear twice. PBS was applied in the negative control group. Then the mice were killed with their 2 ears cut off. Microscopy was used to measure the right ear swelling degree, and differences in weight and thickness between the 2 ears. ELISA was used to detect the levels of IL-2 and INF-gamma in the ear tissues.

RESULTS

The transdermal permeation rates of Z-VAD-FMK at the time points 6, 12, and 24 h were 2.15%, 9.62%, and 12.85% respectively. The right ear swelling degree, and differences in weight and thickness between the 2 ears in the 2.5 mmol/L Z-VAD-FMK group were (12.5 +/- 1.4) x 10(-2) mm, (3.2 +/- 0.3) mg, and (11.8 +/- 1.3) x 10(-2) mm respectively, all significantly lower than those of the negative control group [(19.1 +/- 2.0) x 10(-2) mm, (4.3 +/- 0.4) mg, and (16.8 +/- 1.7) x 10(-2) mm, all P < 0.05]. The IL-2 and INF-gamma levels in the ear skin lesion of the 2.5 mmol/L Z-VAD-FMK group were (148 +/- 10) and (650 +/- 45) pg/ml respectively, both significantly lower than those of the negative control group [(205 +/- 18) and (1030 +/- 58) pg/ml, both P < 0.05].

CONCLUSION

Z-VAD-FMK can permeate through mouse skin and inhibit the activation and proliferation of T lymphocytes, leading to the inhibitory effect of contact allergic reaction.

摘要

目的

研究广谱半胱天冬酶抑制剂N-苄氧羰基-Val-Ala-Asp-氟甲基酮（Z-VAD-FMK）对过敏性接触性皮炎（ACD）的影响。

方法

处死一只Balb/c小鼠，剥取其背部皮肤，放入Franz扩散池，涂抹Z-VAD-FMK。采用高效液相色谱法检测渗透速率。将二硝基氟苯（DNFB）涂抹于脱毛后的小鼠背部作为诱导剂，6天后再涂抹于右耳背侧作为激发剂，以建立小鼠ACD模型。激发后2小时，将24只小鼠模型分为4组，每组6只，分别在右耳背侧涂抹浓度为0.1、0.5、2.5和5 mmol/L的Z-VAD-FMK，每日2次。阴性对照组涂抹PBS。然后处死小鼠，剪下双耳。用显微镜测量右耳肿胀程度以及双耳重量和厚度的差异。采用酶联免疫吸附测定法（ELISA）检测耳组织中白细胞介素-2（IL-2）和γ干扰素（INF-γ）水平。

结果

Z-VAD-FMK在6、12和24小时的透皮渗透率分别为2.15%、9.62%和12.85%。2.5 mmol/L Z-VAD-FMK组的右耳肿胀程度以及双耳重量和厚度的差异分别为（12.5±1.4）×10⁻² mm、（3.2±0.3）mg和（11.8±1.3）×10⁻² mm，均显著低于阴性对照组[（19.1±2.0）×10⁻² mm、（4.3±0.4）mg和（16.8±1.7）×10⁻² mm，P均<0.05]。2.5 mmol/L Z-VAD-FMK组耳皮肤病变组织中IL-2和INF-γ水平分别为（148±10）和（650±45）pg/ml，均显著低于阴性对照组[（205±18）和（1030±58）pg/ml，P均<0.05]。