[关键词]
[摘要]
目的 制备双氢青蒿素脂质体(DHA-LPs),并评价其外用治疗银屑病的作用。方法 采用薄膜分散法制备DHALPs,利用单因素试验结合Box-Behnken设计-响应面法(BBD-RSM)优化工艺参数,采用透射电子显微镜(TEM)、动态透析法及稳定性考察评价其理化性质。采用咪喹莫特(IMQ)诱导的小鼠银屑病样皮损模型,结合HE染色、血常规分析、ELISA分析评价DHA-LPs的抗银屑病作用。结果 DHA-LPs确定的最佳制备工艺为:二氯甲烷为溶剂、药脂比1∶4.329、胆固醇与磷脂比1∶1.833、水化温度46℃。测得DHA-LPs粒径(97.39±0.50)nm、多分散系数(0.275±0.002)、ζ电位(-14.52±0.08)mV、包封率(76.03±0.47)%,载药量(31.21±1.53)%。TEM显示DHA-LPs呈椭球形,体外释放呈双相模式,4℃储存28 d稳定性良好。动物药效学结果显示,与DHA溶液相比,DHA-LPs显著缓解IMQ诱导的小鼠银屑病样皮损,逆转IMQ诱导的小鼠脾脏系数升高,改善IMQ诱导的表皮结构、脾脏结构及中性粒细胞和单核细胞的异常,降低血清中促炎因子[白细胞介素(IL)-6、IL-17、IL-23、肿瘤坏死因子-α(TNF-α)]水平,升高小鼠血清中抑炎因子[转化生长因子-β(TGF-β)]水平。结论 双氢青蒿素外用缓解小鼠银屑病样皮损,将其制备成脂质体制剂能够增强其抗银屑病作用,拓展了青蒿素类药物的应用范畴,为银屑病的治疗提供了一种新的策略。
[Key word]
[Abstract]
Objective To prepare dihydroartemisinin liposomes (DHA-LPs) and evaluate their efficacy in the treatment of psoriasis when applied topically.Methods DHA-LPs were prepared using the thin film dispersion method. The process parameters were optimized by single-factor experiments combined with Box-Behnken design-response surface methodology (BBD-RSM). The physicochemical properties of DHA-LPs were evaluated by transmission electron microscopy (TEM), dynamic dialysis, and stability studies. The antipsoriasis effect of DHA-LPs was evaluated using an imiquimod (IMQ)-induced psoriasis-like skin lesion model in mice, combined with HE staining, blood routine analysis, and ELISA analysis.Results The optimal preparation process of DHA-LPs was determined as follows: dichloromethane as the solvent, drug-to-lipid ratio of 1∶4.329, cholesterol-to-phospholipid ratio of 1∶1.833, and hydration temperature of 46 ℃. The particle size of DHA-LPs was (97.39 ± 0.50) nm, the polydispersity index was (0.275 ± 0.002), the ζ potential was (-14.52 ± 0.08) mV, the entrapment efficiency was (76.03 ± 0.47)%, and the drug loading was (31.21 ± 1.53)%. TEM showed that DHA-LPs were ellipsoidal in shape, and the in vitro release followed a biphasic pattern. The stability of DHA-LPs was good after 28 days of storage at 4 ℃. The animal pharmacodynamics results showed that compared with DHA solution, DHA-LPs significantly alleviated IMQ-induced psoriasis-like skin lesions in mice, reversed the increase in spleen coefficient induced by IMQ, improved the abnormal epidermal and splenic structures and neutrophile granulocytes and monocytes induced by IMQ, and reduced the levels of pro-inflammatory factors [(Interleukin (IL)-6, IL-17, IL-23, Tumor necrosis factor-α (TNF-α)] in serum while increasing the level of anti-inflammatory factor [Transforming growth factor-β (TGF-β)] in mouse serum.Conclusion Topical application of dihydroartemisinin can alleviate psoriasis-like skin lesions in mice. Preparing it into liposomal formulation can enhance its antipsoriasis effect, expanding the application scope of artemisinin derivatives and providing a new strategy for the treatment of psoriasis.
[中图分类号]
R283.6
[基金项目]
国家自然科学基金资助项目(82173767);山西省基础研究计划(自由探索类)青年科学研究项目(202403021212250);山西省博士启动基金资助项目(SD2420);山西医科大学校级博士启动基金资助项目(XD2313)