玉凝膏及其拆方治疗痤疮的量效关系研究

Dose-effect Relationship of Yuning Ointment and its Decomposed Recipes on Acne in Mice

目的:探讨玉凝膏及其拆方防治油酸诱导小鼠痤疮的量效。方法:油酸涂抹于小鼠背部2 cm×2 cm范围,每天1次,连续21 d,建立痤疮模型。于d22,小鼠背部分别涂抹梯度浓度地乌生药量(1.06~1 060.23 mg·kg^(-1)·d^(-1),k=0.316),玉凝膏去地乌生药量(4.73~1 767.75 mg·kg^(-1)·d^(-1),k=0.316),玉凝膏生药量(2.84~2 827.28 mg·kg^(-1)·d^(-1),k=0.316),连续14 d。苏木精-伊红(HE)染色观察皮肤病理变化,形态计量皮脂腺直径、毛囊导管角化面积比;酶联免疫吸附法检测血清白细胞介素(IL)-1、IL-6和肿瘤坏死因子(TNF)-α水平;Graphpad Prism 5软件非线型可变斜率回归不同药物的半数有效剂量(ED_(50)),确定方剂量效。结果:各给药组防治痤疮呈剂量依赖趋势,量效曲线呈"S"型。玉凝膏及其拆方显著缓解皮脂腺肥大与毛囊导管角化,并降低血清IL-1、IL-6和TNF-α水平;其中玉凝膏组疗效最优。以地乌剂量为标识进行回归,结果显示玉凝膏组对皮脂腺肥大改善作用的ED_(50)为地乌组的0.28倍;对毛囊导管角化抑制作用的ED_(50)为地乌组的0.14倍;抑制IL-1,IL-6与TNF-α分泌作用的ED50分别为地乌组的0.15,0.49与0.24倍。以玉凝膏去地乌方剂剂量为标识进行回归,结果显示在改善皮肤病理变化与抑制细胞因子分泌的作用上,玉凝膏组ED_(50)均小于玉凝膏去除地乌的ED_(50)。结论:玉凝膏通过调节免疫防治痤疮,且方剂配伍后可增效地乌。

Objective： To study the dose-effect relationship of Yuning ointment and its decomposed recipes in the treatment of oleic acid induced acne in mice. Methods： Oleic acid was administrated to the back（ 2 cm ×2 cm） of the mice（ once a day） for 21 days to induce acne. At d22,the gradient dosage of Anemone flaccida crude drug（ 1. 06-1 060. 23 mg·kg-1·d-1,k=3. 16）,Yuning ointment without Anemone flaccida crude drug（ 4. 73-1 767. 75 mg·kg-1·d-1,k = 3. 16） and Yuning ointment（ 2. 84-2 827. 28 mg·kg-1·d-1,k=3. 16） was respectively administrated to the back of mice for 14 days. The pathological changes of skin were observed by hematoxylin-eosin（ HE） staining. The diameter of sebaceous glands and the ratio of follicular keratinization area were morphometrically analyzed. The serum levels of IL-1,IL-6 and TNF-α were detected by ELISA assay. The median effective dosages（ ED50） of Anemone flaccida in the three prescriptions were regressed by Prism 5. 01 software to determine the prescription dose-effect. Results： All the therapy groups were with significantly relieved pathological changes of sebaceous glands hypertrophy and follicular keratinization,and decreased serum levels of IL-1,IL-6 and TNF-α in a dose-dependent manner. The dose-response curves showed an＂S＂shape. Among the three therapy groups,the effect of Yuning ointment was the best. The ED50 of Yuning ointment regressed by Anemone flaccida dose was 0. 28-fold for improving sebaceous glands hypertrophy,0. 14-fold for inhibiting follicular keratinization,and 0. 15-,0. 49-and0. 24-fold for decreasing serum levels of IL-1,IL-6 and TNF-α.. Regressed by Yuning ointment without Anemone flaccida,the ED50 of Yuning ointment was lower than Yuning ointment without Anemone flaccid in terms of improving pathological changes and inhibiting the secretion of cytokines. Conclusion： Yuning ointment can prevent and treat acne through regulating immune function. And the prescription compatibility can enhance the effects of Anemone flaccida.