Qin Zhi-Jian, Yan Qiao, Hang Ling-Yu, Tang Xiao-Han, Li Fang-Qin, Xue Yu-Ye, Yuan Hai-Long
School of Pharmacy, Anhui Medical University Hefei 230032, China.
Department of Pharmacy,Air Force Medical Center, PLA Beijing 100142, China.
Zhongguo Zhong Yao Za Zhi. 2023 Apr;48(8):2116-2125. doi: 10.19540/j.cnki.cjcmm.20221223.301.
This study aims to separate and characterize self-assembled nanoparticles(SAN) from Shaoyao Gancao Decoction(SGD) and determine the content of active compounds. Further, we aimed to observe the therapeutic effect of SGD-SAN on imiquimod-induced psoriasis in mice. The separation of SGD was performed by dialysis, and the separation process was optimized by single factor experiment. The SGD-SAN isolated under the optimal process was characterized, and the content of gallic acid, albiflorin, paeoniflorin, liquiritin, isoliquiritin apioside, isoliquiritin, and glycyrrhizic acid in each part of SGD was determined by HPLC. In the animal experiment, mice were assigned into a normal group, a model group, a methotrexate group(0.001 g·kg(-1)), and SGD, SGD sediment, SGD dialysate, and SGD-SAN groups of different doses(1, 2, and 4 g·kg(-1)) respectively. The psoriasis grade of mice was evaluated based on the pathological changes of skin lesions, the content of inflammatory cytokines, organ index and other indicators. The results showed that SAN obtained by centrifugation at 13 000 r·min~(-1) for 30 min was stable after dialysis for 4 times, which were uniform spherical nanoparticles with the particle size of(164.43±1.34) nm, the polydispersity index of(0.28±0.05), and the Zeta potential of(-12.35±0.80) mV. The active compound content accounted for more than 70% of SGD. Compared with the model group, SAN and SGD decreased the skin lesion score, spleen index, and inflammatory cytokine levels(P<0.05 or P<0.01) and alleviated the skin thickening and infiltration of inflammatory cells. However, the sediment group and the dialysate group had no obvious effect. SGD showed a good therapeutic effect on imiquimod-induced psoriasis in mice, and SAN demonstrated the effect equivalent to SGD in a dose-dependent manner. Therefore, we conclude that the SAN formed during decocting is the main active form of SGD, which can lower the levels of inflammatory cytokines, promote the normal differentiation of keratinocytes, and reduce the infiltration of inflammatory cells in the treatment of psoriasis lesions in mice.
本研究旨在从芍药甘草汤（SGD）中分离并表征自组装纳米颗粒（SAN），并测定活性成分的含量。此外，我们旨在观察SGD - SAN对咪喹莫特诱导的小鼠银屑病的治疗效果。通过透析对SGD进行分离，并通过单因素实验优化分离过程。对在最佳工艺条件下分离得到的SGD - SAN进行表征，并采用高效液相色谱法测定SGD各部分中没食子酸、白花芍药苷、芍药苷、甘草苷、异甘草苷芹糖糖苷、异甘草苷和甘草酸的含量。在动物实验中，将小鼠分别分为正常组、模型组、甲氨蝶呤组（0.001 g·kg⁻¹）以及不同剂量（1、2和4 g·kg⁻¹）的SGD、SGD沉淀组、SGD透析液组和SGD - SAN组。根据皮肤病变的病理变化、炎性细胞因子含量、器官指数等指标评估小鼠的银屑病等级。结果表明，在13000 r·min⁻¹离心30 min获得的SAN经4次透析后稳定，为均匀的球形纳米颗粒，粒径为（164.43±1.34）nm，多分散指数为（0.28±0.05），ζ电位为（ - 12.35±0.80）mV。活性成分含量占SGD的70%以上。与模型组相比，SAN和SGD降低了皮肤病变评分、脾脏指数和炎性细胞因子水平（P<0.05或P<0.01），减轻了皮肤增厚和炎性细胞浸润。然而，沉淀组和透析液组无明显效果。SGD对咪喹莫特诱导的小鼠银屑病显示出良好的治疗效果，且SAN以剂量依赖的方式表现出与SGD相当的效果。因此，我们得出结论，煎煮过程中形成的SAN是SGD的主要活性形式，其在治疗小鼠银屑病皮损时可降低炎性细胞因子水平，促进角质形成细胞的正常分化，并减少炎性细胞浸润。
