青蒿素对急性皮炎的防治研究

Prevention and treatment of acute dermatitis by artemisinin： experiment with mice

目的探讨青蒿素（Art）防治急性皮炎的作用机制。方法制造BALB／C小鼠和C57BL／6小鼠的混合淋巴细胞悬液。体外培养，分对照组，和不同浓度Art组，计算不同给药浓度下的淋巴细胞刺激指数。小鼠淋巴细胞悬液接种于96孔板，加入相应浓度的Art，用MTF法检测对各组细胞存活抑制率。60只ICR小鼠均分为6组，除对照组外，分别用基质，地塞米松（Dex）和高、中、低浓度Art涂左耳，再涂二甲苯致鼠耳廓肿胀。30min后处死小鼠，左右耳片厚度差为肿胀度。耳片组织匀浆，用双抗体夹心ELISA法检测IL-6含量。结果小鼠混合淋巴细胞培养48h后刺激指数最高可达（1．68±0．084），而Art组刺激指数均较低。Art浓度分别为5μmol／L和200μmol／L时，淋巴细胞刺激指数均显著低于Dex组（分别为P〈0．05和P〈0．01）。Art作用小鼠淋巴细胞24、48、72hA，细胞存活率都在50％以上，Art浓度（200μmol/L）时对细胞的抑制分别为（23．35±2．18）％、（29．60±2．69）％和（45．32±5．23）％。Art剂量依赖性地抑制二甲苯刺激所致的小鼠耳肿胀度。模型组IL-6表达量为[（256±7．2）ng／L]，显著高于正常组[（185±14．3）ng／L，P〈0．011。不同浓度Ar组的IL／-6水平均低于模型组，此作用呈剂量依赖性。结论Art是一种高效低毒的新型免疫调节剂，对于急性皮炎具有较好的防治作用，其机制可能与抗炎、抗过敏以及免疫调节作用有关。

Objective To study the immune effects and mechanism of artemisinin （Art）, extracted from traditional Chinese medicinal herb, on acute dermatitis. Methods Mixed lymohocyte suspension of BALB/C and C57BL/6 mice was made, cultured in vitro, and divided into control group and group with Art at different concentrations, stimulation index （SI） of the lymphocytes were calculated. Another mixed lymohocyte suspension was inoculated into a 96-well plate and added with Art at different concentrations. 3-（4, 5-dimethylthiazol-2-yl）-2,5-diphenyl- 2h-tetrazolium bromide （MTT） method was used to examine the survival inhibition rates of the lymphocyte. Sixty ICR mice were divided into 6 equal groups to undergo rubbing of the left ear with matrix, dexamethasone （Dex）, or Art at the concentrations of 2%, 4%, and 8% respectively, and then rubbing of xylene to cause swelling except the control group. Thirty minutes later the mice were killed with pieces of ear conch cut down. The difference in the thichness of both ear conches were calculkated as the degree of swelling. Tissue homogenate of ear pieces was made and the supernatant was collected. Double antibody ELISA sandwich method was used to examine the IL-6 level therein. Results The SI of the mice lymphocytes after 48-hour culture reached （1.68 ~ 0.084）, and the SI values of the Art groups with different concentrations were all lower than those of the corresponding Dex control groups dose-dependently （P 〈0.05 for the Art 5 μmol/L group and P 〈0.01for the Art 10-200μ mol/L groups）. MTT method showed that the lymphocyte survival rates 24, 48, and 72 hours after culture were all over 50% for the Art groups at different concentrations, and were （23.35 ±2.18） %, （29.60 ± 2.69） %, and （45.32 ± 5.23） % respectively when the Art concentration was 200 Ix mol/L. The highest inhibition rate of the positive Dex control group was （69.97 ±6.18） %, significantly higher than that of any Art groups （all P〈0.05）. The ear piece swelling degrees of different concentration Art groups were all milder than that of all other groups dose-dependently. The IL-6 level of the model group was （256±7.2）ng/L, signigficantly higher than that of the normal control group [（185±14.3） ）ng/L, P〈0.01]. And the IL-6 levels of the Art groups were all significantly lower than that of the model group dose-dependently （all P〈0.01）. Conclusion Highly effective and with low toxicity eanbe use to prevcent and eure acute dermatitis. Its pharmacological mechanism may be related to its anti-inflammatory, anti-allergic, and immunomodulatory effects.