Li Hui-Zhu, Song Dan-Dan, Chang Chen-Qi, Lu Chang, Liu Yue, Chen Rui
School of Pharmacy,Nanjing University of Chinese Medicine Nanjing 210023,China.
Zhongguo Zhong Yao Za Zhi. 2023 Aug;48(16):4394-4401. doi: 10.19540/j.cnki.cjcmm.20230510.301.
This study focused on the separation, characterization, content determination, and antiviral efficacy research on colloidal particles with different sizes in Maxing Shigan Decoction(MXSG). The mixed colloidal phase of MXSG was initially separated into small colloidal particle segment(S), medium colloidal particle segment(M), and big colloidal particle segment(B) using ultrafiltration. Further fine separation was performed using size-exclusion chromatography. Dynamic light scattering(DLS) and transmission electron microscopy(TEM) were employed to characterize the size and morphology of the separated colloidal particles. UPLC-MS/MS was used to determine the content of ephedrine, amygdalin, glycyrrhizic acid, and the EDTA complexometric titration was used to measure the calcium(Ca(2+)) content in different colloidal phases. Finally, a respiratory syncytial virus(RSV) infection mouse model was established using intranasal administration. The experimental groups included a blank group, a model group, a ribavirin group, an MXSG group, an S group, an M group, and a B group. Oral administration was given for treatment, and pathological changes in mouse lung tissue and organ indices were evaluated. The results of the study showed that the distribution of ephedrine, amygdalin, glycyrrhizic acid, and Ca(2+) content was not uniform among different colloidal segments. Among them, the B segment had the highest proportions of the three components, except for Ca(2+), accounting for 46.35%, 53.72%, and 92.36%, respectively. Size-exclusion chromatography separated colloidal particles with uniform morphology in the size range of 100-500 nm. Compared to the S and M segments, the B segment showed an increased lung index inhibition rate(38.31%), spleen index, and thymus index in RSV-infected mice, and it improved the infiltration of inflammatory cells and lung injury in the lung tissue of mice. The complex components in MXSG form colloidal particles of various sizes and morphologies through heating, and small-molecule active components such as ephedrine, amygdalin, glycyrrhizic acid, and Ca(2+) participate in the assembly to varying degrees. The main material basis for the antiviral effect of MXSG is the colloidal particles with certain particle sizes formed by the assembly of active components during the heating process.
本研究聚焦于麻杏石甘汤（MXSG）中不同大小胶体颗粒的分离、表征、含量测定及抗病毒功效研究。首先利用超滤将MXSG的混合胶体相分离为小胶体颗粒段（S）、中胶体颗粒段（M）和大胶体颗粒段（B）。进一步采用尺寸排阻色谱进行精细分离。运用动态光散射（DLS）和透射电子显微镜（TEM）对分离得到的胶体颗粒的大小和形态进行表征。采用超高效液相色谱-串联质谱（UPLC-MS/MS）测定麻黄碱、苦杏仁苷、甘草酸的含量，并用乙二胺四乙酸络合滴定法测定不同胶体相中的钙（Ca²⁺）含量。最后，通过滴鼻给药建立呼吸道合胞病毒（RSV）感染小鼠模型。实验组包括空白组、模型组、利巴韦林组、MXSG组、S组、M组和B组。给予口服给药进行治疗，并评估小鼠肺组织的病理变化和器官指数。研究结果表明，麻黄碱、苦杏仁苷、甘草酸和Ca²⁺含量在不同胶体段中的分布并不均匀。其中，除Ca²⁺外，B段中这三种成分的比例最高，分别占46.35%、53.72%和92.36%。尺寸排阻色谱分离出了形态均匀、大小在100 - 500 nm范围内的胶体颗粒。与S段和M段相比，B段在RSV感染小鼠中表现出更高的肺指数抑制率（38.31%）、脾脏指数和胸腺指数，并改善了小鼠肺组织中炎性细胞的浸润和肺损伤。MXSG中的复杂成分通过加热形成了各种大小和形态的胶体颗粒，麻黄碱、苦杏仁苷、甘草酸和Ca²⁺等小分子活性成分不同程度地参与了组装。MXSG抗病毒作用的主要物质基础是活性成分在加热过程中组装形成的具有一定粒径的胶体颗粒。
