地乌配伍增效加味逍遥丸治疗皮肌炎的量效研究

The Dose-effect Relationship of the Compatible Enhancement of Anemone Flaccida and Jiaweixiaoyao Pills in Treating Dermatomyositis

目的:探讨地乌配伍增效加味逍遥丸治疗大鼠皮肌炎的量效关系。方法:膜抗原与毒素诱导建立大鼠皮肌炎模型(膜抗原d0,d7致敏,d28,d31,d35,d38发敏;d56给药二氢睾酮、右旋糖酐铁、多黏菌素),灌胃给予梯度剂量地乌生药2.81~27 919.42 mg·kg-1·d-1,k=3.16,加味逍遥丸生药(1.83~18 173.79 mg·kg-1·d-1,k=3.16),加味逍遥丸加地乌生药(2.11~20 969.76 mg·kg-1·d-1,k=3.16)。苏木精-伊红(HE)染色观察肌肉病理变化,形态计量T淋巴细胞浸润体积比;钙化结节染色观察皮肤钙化,形态计量钙化结节体积比;免疫组化显示肌肉组织foxp3及IL-17RA表达并形态计量其阳性表达量;各指标用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(ED50),确定量效关系,观察地乌对加味逍遥丸方剂的配伍增效作用。结果:各给药组防治皮肌炎呈剂量依赖趋势,量效曲线呈"S"型。其中方剂加地乌组疗效最优,以加味逍遥丸剂量为标识进行回归,方剂加地乌组在逆转T淋巴细胞浸润体积比、钙化结节体积比、IL-17RA表达量、Foxp3表达量的ED50分别是方剂组的0.43倍、0.72倍、0.71倍、0.68倍;以地乌剂量为标识,方剂加地乌组改善各病变ED50分别是方剂组的0.70倍、0.44倍、0.74倍、0.81倍。结论:加味逍遥丸能有效治疗大鼠皮肌炎,地乌具有配伍增效该方剂的作用。

Objective: To study the dose-effect relationship of the compatible enhancement of Anemone flaccida to Jiaweixiaoyao pills in treating dermatomyositis( DM). Methods: Nighty SD rats were randomly divided into 5 groups,the rats in four groups were administrated with membrane antigen( once weekly from d00 to d07 for sensitization,and twice weely from d28 to d38 for challenge),at d63,the rats were administrated with dihydrotestosterone( DHT),Fe-dextrin( Fe-Dex),and polymyxin( PMX) to establish the rat DM model. At d21,the rats were treated with gradient dosages of Anemone flaccida and Jiaweixiaoyao pills( 1.83-18173.79 mg·kg-1·d-1,k= 3.16),Anemone flaccida( 0.28-2795.97 mg·kg-1·d-1,k = 3.16),Jiaweixiaoyao pills and Anemone flaccida( 2.11-20969.76 mg·kg-1·d-1,k= 3.16),respectively.The pathological features were observed after hematoxylin-eosin staining.The volume ratio of infiltrated lymphocytes in skeletal muscles or arteriole calcified nodules in subcutaneous tissues were analyzed with morphometry. The expression of foxp3 and IL-17 RA were detected by IHC. The ED50 was calculated by non-linear regression with various slope using Prism-5.0 software. Results: The effects of Jiaweixiaoyao pills and Jiaweixiaoyao pills plus Anemone flaccida on DM showed a dose-dependent tendency.The dose-response curves showed"S"shape.The efficacy of Jiaweixiaoyao pills on rat DM was the most significant among the three formulas. The ED50 of Jiaweixiaoyao pills plus Anemone flaccida regressed by Jiaweixiaoyao pills dose was 0.63-fold for decreasing the T lymphocytes infiltration,0.72-fold for decreasing the arteriole calcified nodules,0.71-fold for inhibiting the expression of IL-17 RA,and 0.68-fold for increasing the expression of foxp3. The ED50 of Jiaweixiaoyao pills plus Anemone flaccida regressed by Anemone flaccida dose was 0.70-fold,0.44-fold,0.74-fold and 0.81-fold for T lymphocytes infiltration,the arteriole calcified nodules,the expression of IL-17 RA and the expression of foxp3. Conclusion: Jiaweixiaoyao pills can prevent and treat rat DM effectively. Anemone flaccida has obvious compatibility synergy in the whole formula of enhancing the therapeutic effect and reducing toxicity.