茯苓硫酸酯化多糖的制备及其核磁共振波谱分析

[目的]用不同方法制备3种茯苓多糖硫酸酯（Pachyman sulfate,PS）,并对其进行核磁共振波谱对比分析,为多糖硫酸化提供基础数据。[方法]以氯磺酸为磺化试剂硫酸化茯苓多糖制备茯苓多糖硫酸酯PS—I和PSII,以氨基磺酸为磺化试剂制备PS-Ⅲ,分析磺化反应前后和不同硫酸根取代度（Degree of sulfation,DS）的多糖碳原子化学位移的变化规律。[结果]PS—I（DS=2．80）分子所有自由羟基均发生取代反应;PS—H（DS=1．55）C-6羟基被全部酯化,C-2和C-4羟基发生部分酯化反应;PS-Ⅲ（DS=0．95）C-6和C-2羟基发生部分硫酸酯化反应。[结论]发生取代反应的碳原子化学位移移动的距离与PS的DS呈正相关,核磁共振技术能够提供多糖结构解析的关键数据。     

Effect of pachyman polysaccharides on Th17/Treg balance in systemic lupus erythematosus patients

AIM: To investigate the immunomodulatory effect of pachyman polysaccharides (PPS) on T helper 17 cell (Th17)/regulatory T cell (Treg) balance in the peripheral blood of systemic lupus erythematosus (SLE) patients. METHODS: The CD4⁺ T cells were isolated from the peripheral blood samples obtained from 45 SLE patients and 35 healthy controls enrolled in our study using magnetic bead separation method. The proportions of Th17 and Treg cells were measured by flow cytometry. The CD4⁺ T cells from SLE patients and healthy controls were treated with PPS. The cytoto-xicity of PPS was evaluated by detecting cell viability with MTT assay. The contents of interleukin-17 (IL-17), IL-6, IL-10 and transforming growth factor-β (TGF-β) were measured by ELISA. The expression of retinoid-related orphan receptor γt (RORγt) and forkhead box protein P3 (Foxp3) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTS: The Th17 cells were significantly elevated, while Treg cells were obviously decreased in the SLE patients compared with the healthy control group (P<0.05). Compare with control group, the contents of IL-17 and IL-6 were decreased, while the contents of IL-10 and TGF-β were increased (P<0.05). The expression of RORγt at mRNA and protein levels was down-regulated and the expression of Foxp3 was up-regulated (P<0.05). The ratio of Th17/Treg was decreased in 100 μg/L nontoxic PPS-treated CD4⁺ T cells isolated from the SLE patients (P<0.05). CONCLUSION: PPS treatment inhibits Th17 cells and elevates Treg cells in the CD4⁺ T cells isolated from SLE patients, which may have a therapeutic effect on SLE patients.     

微波辅助酶法提取茯苓中茯苓多糖的工艺研究

[目的]提高茯苓中水溶性多糖的提取率,寻求最优的提取条件。[方法]利用均匀设计试验对微波辅助酶法提取的主要影响因素进行优化。[结果]确定了微渡辅助酶法的最佳工艺条件为微波提取时间17 min、功率550 W、固液比1∶30、提取次数2次、酶解温度60℃、酶解时间120 min、加酶量0.5%、pH值6.0,在此条件下,茯苓水溶性粗多糖的提取率为3.58%。[结论]微波辅助酶法为茯苓水溶性多糖提取的有效方法。     

九资河茯苓多糖的超声波提取工艺及抑菌性研究

利用超声波技术,最大程度的提取茯苓多糖,并用苯酚-硫酸法测定其含量,优选茯苓多糖的最佳超声提取工艺。首先,对茯苓多糖提取的超声功率、提取时间和料液比进行单因素考察;在单因素实验的基础上,利用响应面设计试验,以茯苓多糖得率为指标,得出最佳超声水提工艺条件为：超声功率120 W,提取时间27 min,料液比1：38,利用牛津杯法来探究茯苓多糖的抑菌性,在抑菌性研究中,茯苓多糖对枯草芽孢杆菌和金黄色葡萄球菌有显著的抑菌性。     

羧甲基茯苓聚糖制备新工艺

该工艺采用两次加碱法以水－乙醇为介质制备羧甲基茯苓聚糖,可提高氯乙酸钠的利用率。本文着重研究了羧甲基化反应条件对取代度和绝对粘度的影响,用Ｘ－衍射,ＩＲ,＾１３ＣＮＭＲ对ＣＭＰ进行了结构表征。     

茯苓活性成分提取工艺优化

为了优化茯苓有效成分的提取工艺,分别以茯苓酸和茯苓多糖含量为考核指标,在单因素试验确定因素和水平的基础上,采用正交试验法分别对茯苓酸和茯苓多糖提取工艺进行考察。优选茯苓酸的最佳提取工艺为采用6倍药材质量的85%乙醇在55℃条件下提取1 h,优选茯苓多糖的最佳提取工艺为采用10倍药材质量的水提取3次,每次2 h。优选的提取工艺合理、稳定、可行,为进一步工业化生产提供科学的理论依据。