摘要
目的:探究茯苓多糖(PCP)对脾虚大鼠免疫功能和肠道菌群的调节作用.方法:采用超声提取和大孔树脂纯化的方法提取纯化PCP,并进行初步的结构解析.采用"饮食不节+劳倦过度法"构建脾虚大鼠模型,模拟肠道菌群紊乱和免疫低下的机体状态,造模结束后灌胃PCP(75、150、300mg/kg)干预,通过酶联免疫吸附试验、免疫组化、16S rRNA基因高通量测序等方法,检测脾虚大鼠的免疫功能和肠道菌群结构及丰度.结果:PCP主要是由阿拉伯糖、甘露糖、半乳糖和葡萄糖构成的多孔疏松的β-葡聚糖.PCP可有效提高脾虚大鼠的体质量,显著增加脾虚大鼠的脏器指数,促进血清中免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)、干扰素-γ(IFN-γ)的分泌和表达.肠道菌群分析表明,在门水平上,PCP显著增加脾虚大鼠肠道菌群的拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)的比值.在属水平上,PCP显著降低肠球菌属(Enterococcus)丰度,PCP高剂量组显著增加免疫相关的普雷沃氏菌属(Prevotellaceae_Unclassified、Prevotellaceae_UCG-003、Prevotella_9)丰度,恢复脾虚大鼠的肠道菌群稳态.结论:PCP可以有效提高脾虚大鼠的免疫功能,其途径可能与肠道菌群有关.
Abstract
Objective:To study the effect of Poria cocos polysaccharide on immune function and intestinal flora regulating of spleen deficiency rats.Methods:PCP was extracted and purified by ultrasonic extraction and macroporous resin,and its structure was preliminarily analyzed.The rat model of spleen deficiency was established by improper diet and overwork to simulate the body state of intestinal flora disorder and low immunity.Then,PCP(75,150,300 mg/kg)was intragastric gavage to the model rats.The immune function,intestinal flora structure and abundance of spleen deficiency rats were detected by enzyme-linked immunosorbent assay,immunohistochemistry,16S rRNA gene high-throughput sequencing and other methods.Results:PCP was a porous polysaccharide mainly composed of arabinose,mannose,galactose,and glucose.PCP could effectively improve the body mass of rats with spleen deficiency,significantly increase the organ index of rats,and promote the level of IL-2,IL-4,IFN-γ,IgM,and IgG in serum.Gut flora analysis showed that PCP significantly increased the ratio of Firmicutes to Bacteroides at the phylum level.At the genus level,PCP significantly reduced the level of Enterococcus,significantly increased the level of immune-related Prevotellaceae_Unclassified,Prevotellaceae_UCG-003,Prevotella_9,and restored the structure and abundance.Conclusion:PCP can effectively improve the immune function of rats with spleen deficiency,which may be related to the intestinal flora.
基金项目
国家自然科学基金青年科学基金项目(82204748)
中华中医药学会青年人才托举工程项目(CACM-2022-QNRC2-B03)
国家重点研发计划项目(2017YFC1701605)
安徽省菌类药物研发"115"产业创新团队项目(皖人才[2020]4号)
安徽省中央引导地方科技发展专项(201907d07050002)
安徽省教育厅项目(2022AH050512)
安徽省科技攻坚计划项目(202423110050002)
NETL
NSTL
万方数据