荧光内窥式共聚焦显微镜下青藤碱固体脂质纳米粒注射治疗类风湿性关节炎的研究  

Treatment of rheumatoid arthritis by injection of sinomenine solid lipid nanoparticles under a fluorescence endoscopic laser confocal microscope

作　　者：张筠昊 文武龙 张炜烨 梁霄 孙鑫 肖展 赵玉强 李安琪 赵佩然 白皓天 杨婧[2] 王锐[1,3] ZHANG Jun-hao;WEN Wu-long;ZHANG Wei-ye;LIANG Xiao;SUN Xin;XIAO Zhan;ZHAO Yu-qiang;LI An-qi;ZHAO Pei-ran;BAI Hao-tian;YANG Jing;WANG Rui(School of Pharmacy,Heilongjiang University of Chinese Medicine,Harbin 150040,China;School of Basic Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Key Laboratory of Basic and Applied Research of Northern Medicine,Ministry of Education,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学药学院,黑龙江哈尔滨150040 [2]黑龙江中医药大学基础医学院,黑龙江哈尔滨150040 [3]黑龙江中医药大学,教育部北药基础与应用研究重点实验室,黑龙江哈尔滨150040

出　　处：《中国中药杂志》2023年第14期3786-3792,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81603418,82074271);黑龙江优秀青年人才项目(2020YQ05)。

摘　　要：应用荧光内窥式共聚焦显微镜(fluorescence endoscopic laser confocal microscope,FELCM)引导关节注射青藤碱固体脂质纳米粒(sinomenine solid lipid nanoparticles,Sin-SLN),并考察体外药效评估Sin-SLN治疗类风湿性关节炎(rheumatoid arthritis,RA)的效果。采用乳化蒸发-低温固化法制备Sin-SLN,最佳处方工艺下制备的Sin-SLN包封率为64.79%±3.12%,载药量为3.84%±0.28%,测定平均粒径为(215.27±4.21)nm,Zeta电位为(-32.67±0.84)mV,初步稳定性考察中,Sin-SLN放置30 d后稳定性良好。皮下注射完全弗氏佐剂和卵白蛋白建立兔RA模型,药效实验设对照组、模型组、Sin组(1.5 mg·kg^(-1))、Sin-SLN组(1.5 mg·kg^(-1))和地塞米松组(阳性药,1.0 mg·kg^(-1),ig),对照组和模型组仅穿刺不注射药物,各实验组给予相应药物干预治疗。给药后,每日测量家兔膝关节直径及局部皮肤温度变化,与模型组相比,给药组膝关节直径和局部皮肤温度均下降;使用FELCM观测软骨形态学变化,Sin-SLN组膝关节软骨表面光滑,骨组织结构紧密;采用ELISA测定血清中白细胞介素-1(interleukin-1,IL-1)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达水平,发现Sin-SLN组血清中IL-1和TNF-α表达水平降低;与模型组相比具有显著性差异,苏木素-伊红(hematoxylin-eosin,HE)染色观察滑膜组织形态学变化,Sin-SLN组滑膜组织病变状况得到明显改善。因此,制备的Sin-SLN粒径均匀,性质稳定,通过关节注射给药形成药物贮库,Sin-SLN可有效减轻兔关节肿胀、减轻关节软骨损害,抑制炎症因子表达,抑制关节滑膜上皮增生和炎性细胞浸润,证明Sin-SLN可以有效抑制RA。A fluorescence endoscopic laser confocal microscope(FELCM)was used to direct the injection of sinomenine solid lipid nanoparticles(Sin-SLN)into the joint,and the in vitro effectiveness of Sin-SLN in the treatment of rheumatoid arthritis(RA)was evaluated.Sin-SLN was prepared with the emulsion evaporation-low temperature curing method.The Sin-SLN prepared under the optimal conditions showed the encapsulation efficiency of 64.79%±3.12%,the drug loading of 3.84%±0.28%,the average particle size of(215.27±4.21)nm,and the Zeta potential of(-32.67±0.84)mV.Moreover,the Sin-SLN demonstrated good stability after storage for 30 days.The rabbit model of RA was established by the subcutaneous injection of ovalbumin and complete Freund's adjuvant.Five groups were designed,including a control group,a model group,a Sin(1.5 mg·kg^(-1))group,a Sin-SLN(1.5 mg·kg^(-1))group,and a dexamethasone(positive drug,1.0 mg·kg^(-1),ig)group.The control group and the model group only received puncture treatment without drug injection.After drug administration,the local skin temperature and knee joint diameter were monitored every day.The knee joint diameter and the local skin temperature were lower in the drug administration groups than in the model group(P<0.05,P<0.01).FELCM recorded the morphological alterations of the cartilage of knee joint.The Sin-SLN group showed compact tissue structure and smooth surface of the cartilage.Enzyme-linked immunosorbent assay(ELISA)was employed to determine the serum le-vels of interleukin-1(IL-1)and tumor necrosis factor-α(TNF-α).The findings revealed that the Sin-SLN group had lower IL-1 and TNF-αlevels than the model group(P<0.05,P<0.01).Hematoxylin-eosin(HE)staining was employed to reveal the pathological changes of the synovial tissue,which were significantly mitigated in the Sin-SLN group.The prepared Sin-SLN had uniform particle size and high stability.Through joint injection administration,a drug reservoir was formed.Sin-SLN effectively alleviate joint swelling and cartilage damage of rabbi

关 键 词：荧光内窥式共聚焦显微镜 青藤碱 固体脂质纳米粒 类风湿性关节炎 贮库效应 

分 类 号：R283.6[医药卫生—中药学]