铁皮石斛提取物对小鼠慢性束缚结合睡眠剥夺模型免疫功能的影响

目的 通过网络药理学和动物实验探讨铁皮石斛的茎与花部位提取物对慢性束缚结合睡眠剥夺小鼠模型免疫功能的影响。方法 首先通过网络药理学预测石斛激活免疫细胞的靶点及通路；然后建立慢性束缚结合睡眠剥夺动物模型；采用石斛茎、花提取物(0.5 g/kg)对小鼠进行灌胃给药。31 d造模给药后，检测小鼠体能状态、小鼠自然杀伤(natural killer，NK)细胞、巨噬细胞活性、小鼠外周血中免疫细胞亚群(CD19+CD3e- B细胞、CD19-CD3e+ T细胞、CD4+CD3e+ Th细胞、CD4+CD3e- NK细胞)比例以及小鼠血清中细胞因子(白细胞介素4(interleukin-4，IL-4)、白细胞介素6(interleukin-6，IL-6)和免疫球蛋白G(immunoglobulin G，IgG)水平。结果 筛选出铁皮石斛14个潜在活性成分及 429个作用靶点, 与 884个作用于免疫细胞的靶点取交集得到 125个靶点，通过基因本体(gene ontology, GO) 功能富集分析得到直接参与免疫细胞调控的相关条目 25个，铁皮石斛可能通过促进B细胞、T细胞增殖分化，IL-6受体结合来激活免疫细胞，京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)通路富集析12条，铁皮石斛激活免疫细胞与T细胞受体信号通路、B细胞受体信号通路、细胞周期信号通路等有关。铁皮石斛茎(Dendrobium officinale stem, SDO)、花(Dendrobium officinale flower, FD)组小鼠与模型(Model, MOD)组小鼠比较，跑台力竭时间、脾脏指数、胸腺指数显著提高(P < 0.05或P < 0.01)，血液中NK细胞活性、巨噬细胞活性、CD19+CD3e- B细胞、CD19-CD3e+ T细胞、CD4+CD3e+ Th细胞、CD4+CD3e- NK细胞比例显著上升(P < 0.05或P < 0.01)，血清中IL-4、IL-6、IgG的含量显著上升(P < 0.05或P < 0.01)。结论 铁皮石斛激活免疫细胞具有多成分、多靶点、多通路的特点。铁皮石斛花、茎提取物在慢性束缚结合睡眠剥夺模型中都具有提高小鼠免疫功能的作用。

Effects of Dendrobium officinale extract on immune function of chronic restraint combined with sleep deprivation model in mice

Objective To evaluates the effects of stem and flower extracts from Dendrobium officinale on immune function of chronically restraint and sleep deprived mice model through network pharmacology and animal experiments. Methods Firstly the target and pathway of Dendrobium officinale in activating immune cells were predicted by network pharmacological method. Then the animal model of chronically restraint and sleep deprived mice model was established. The extracts from Dendrobium officinale stem and flower (0.5 g/kg) were administered by gavage to mice. After 31 days modeling and administration, the physical status of mice, the activity of natural killer (NK) cells and macrophages, the percentages of CD19+CD3e- B cells, CD19-CD3e+ T cells, CD4+CD3e+ Th cells and CD4+CD3e- NK cells in peripheral blood, the levels of cytokines interleukin-4 (IL-4), interleukin-6 (IL-6)] and immunoglobulin G (IgG) in serum were measured. Results Fourteen potential active components and 429 action targets of Dendrobium officinale were screened, and 125 targets were obtained by intersection with 884 targets acting on immune cells. Gene ontology (GO) function enrichment analysis showed that 25 related entries were directly involved in immune cell regulation, Dendrobium officinale may activate immune cells by promoting the proliferation and differentiation of B cells and T cells and binding IL-6 receptor and and the enrichment analysis of kyoto encyclopedia of genes and genomes (KEGG) further afforded 12 pathway. The activation of immune cells by Dendrobium officinale was related to T cell receptor signal pathway, B cell receptor signal pathway and cell cycle signal pathway. Compared with the model group, the exhaustion time, spleen index and thymus index of mice in Dendrobium officinale stem and flower group increased significantly (P<0.05 or P<0.01), the activity of NK cells and macrophages, the percentages of CD19+CD3e- B cells, CD19-CD3e+ T cells, CD4+CD3e+ Th cells and CD4+CD3e- NK cells in blood increased significantly (P<0.05 or P<0.01), and the content of IL-4, IL-6 and IgG in serum increased notably (P<0.05 or P<0.01), which was consistent with the predicted results from network pharmacology. Conclusion The immune cells activated by Dendrobium officinale have the characteristics of multi-component, multi-target and multi-channel. The extracts of Dendrobium officinale flower and stem can improve the immune function of chronically restraint and sleep deprived mice model.