透明质酸修饰的盐酸青藤碱脂质体可溶性微针的制备、表征及体外评价  

Preparation,Characterization,and In Vitro Evaluation of Dissolvable Microneedles Loaded with Hyaluronic Acid-modified Sinomenine Hydrochloride Liposome

作　　者：谢询 钟倪俊 莫静媛 黄小玲 邹家祥 张永萍 王利胜[1] XIE Xun;ZHONG Nijun;MO Jingyuan;HUANG Xiaoling;ZOU Jiaxiang;ZHANG Yongping;WANG Lisheng(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;School of Pharmacy,Guizhou University of Tradition Chinese Medicine,Guiyang 550025 Guizhou,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]贵州中医药大学药学院,贵州贵阳550025

出　　处：《中药新药与临床药理》2024年第9期1428-1436,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广州市科技计划项目(202206010188);广州中医药大学“双一流”与高水平大学学科协同创新团队培育项目(2021xk78)。

摘　　要：目的制备一种针尖层负载透明质酸(Hyaluronic acid,HA)修饰的盐酸青藤碱脂质体可溶性微针(HASMH-Lip-DMNs),对其进行表征,并考察其体外透皮性能、细胞摄取能力和体外抗炎能力。方法采用两步浇铸法制备HA-SMH-Lip-DMNs,使用高效液相色谱(HPLC)法测定其载药量,通过扫描电镜、穿刺试验、Franz扩散池法等检测其形态、皮肤穿刺性能和体外透皮能力。以异硫氰酸荧光素脂质体(HA-FITC-Lip/FITCLip)代替HA-SMH-Lip,制备荧光微针(HA-FITC-Lip-DMNs/FITC-Lip-DMNs),并运用流式细胞仪和荧光显微镜观察炎症细胞对HA-FITC-Lip-DMNs/FITC-Lip-DMNs的摄取情况。利用ELISA试剂盒检测细胞培养上清中的炎症因子一氧化氮(NO)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素10(IL-10)水平,评估HA-SMH-Lip-DMNs的抗炎作用。结果制备的HA-SMH-Lip-DMNs形状大小均匀,阵列完整美观,每片平均载药量(114.01±1.04)μg,有良好的穿刺能力。体外透皮结果显示HA-SMH-Lip-DMNs 36 h的累积释放量为(101.47±2.91)μg·cm-2,其透皮能力优于SMH溶液组。体外摄取结果表明RAW 264.7细胞对HA-FITC-Lip-DMNs摄取较多(P<0.01)。与模型组相比,HA-SMH-Lip-DMNs组能显著降低TNF-α、IL-1β和NO的水平(P<0.01),同时提高IL-10的水平(P<0.01)。结论该研究制备的HA-SMH-Lip-DMNs具备良好的形态特征、细胞摄取能力、体外透皮性能以及抗炎活性,有望成为一种新型的经皮给药系统。Objective To prepare a dissolvable microneedle(DMN)with a tip-layer loaded with hyaluronic acid(HA)modified sinomenine hydrochloride liposomes(HA-SMH-Lip),as well as characterize,evaluate its in vitro transdermal permeability,cellular uptake ability,and anti-inflammatory ability.Methods HA-SMH-Lip-DMNs were prepared by a two-step casting method,and the drug loading capacity was determined using HPLC.The morphology,skin permeation properties and in vitro transdermal ability were investigated by scanning electron microscopy,puncture assay and Franz diffusion cell method.Fluorescent microneedles were prepared by replacing HA-SMH-Lip with fluorescein isothiocyanate liposomes(HA-FITC-Lip/FITC-Lip).The uptake behavior of inflammation cells on HA-FITC-Lip-DMNs/FITC-Lip-DMNs was investigated using a flow cytometer and a fluorescence microscope.To evaluate the anti-inflammatory activity of HA-SMH-Lip-DMNs,the levels of inflammatory factors including nitric oxide(NO),tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),and IL-10 in cell supernatants were measured using an ELISA kit.Results The prepared HA-SMH-Lip-DMNs have uniform shape and size,integral and visually pleasing array,and an average drug loading of(114.01±1.04)μg.Additionally,they have good puncture ability.The results of in vitro transdermal experiments showed that the accumulated amounts of HA-SMH-Lip-DMNs were(101.47±2.91)μg·cm-2 at 36 hours.Its transdermal ability was better than that of the SMH solution group and SMH liposome group.In vitro cellular uptake results indicated that HA-FITC-Lip-DMNs were more effectively taken up by RAW 264.7 cells(P<0.01).Compared to the model group,HA-SMH-Lip-DMNs group significantly reduced TNF-α,IL-1β,and NO levels while increase IL-10 levels(P<0.01).Conclusion The prepared HA-SMH-Lip-DMNs have a complete and beautiful morphology with excellent cellular uptake capability,remarkable in vitro transdermal performance,and potent anti-inflammatory properties.HA-SMH-Lip-DMNs are expected to become a new type of trans

关 键 词：盐酸青藤碱 脂质体 透明质酸 可溶性微针 制备工艺 抗炎活性 

分 类 号：R283.6[医药卫生—中药学]