響應(yīng)面法優(yōu)化茯苓多糖硫酸酯化合成工藝及其抗凝血活性分析

陳小紅

摘要：以中藥茯苓（Poria cocos）為原料提取茯苓多糖，以氨基磺酸為酯化試劑對茯苓多糖進行硫酸酯化結(jié)構(gòu)改性，制備硫酸酯化茯苓多糖（SP），在單因素試驗基礎(chǔ)上，應(yīng)用響應(yīng)面法試驗優(yōu)化其制備條件。對獲得的SP進行純化、結(jié)構(gòu)表征，通過小白鼠體內(nèi)抗凝血試驗考察SP的抗凝血活性。結(jié)果表明，最佳的硫酸酯化工藝條件為反應(yīng)溫度85 ℃、反應(yīng)時間4.0 h、氨基磺酸∶多糖質(zhì)量比為3.77∶1，在此條件下，硫酸基團取代度（DS）為1.041，與氯磺酸吡啶法相比取代度相當。精制后SP的紅外圖譜表明，SP具有多糖及硫酸根基團的特征吸收峰;紫外光譜掃描表明SP無核酸、蛋白質(zhì)等雜質(zhì)，純度較高，且有硫酸根基團的特征吸收，硫酸酯化成功。茯苓多糖經(jīng)硫酸酯化結(jié)構(gòu)改性后表現(xiàn)出良好的抗凝血活性，其出血時間、全凝血時間、全凝血時間延長率均明顯提高;SP對凝血酶的抑制作用與其濃度呈正相互作用，隨著SP腹腔注射給藥劑量增大，其全凝血時間、延長率均明顯提高。

關(guān)鍵詞：茯苓（Poria cocos）;硫酸酯化;茯苓多糖;響應(yīng)面法;抗凝血活性

中圖分類號：S567.3+2 ? ? ? ?文獻標識碼：A ? ? ? ?文章編號：0439-8114（2015）23-5995-06

DOI：10.14088/j.cnki.issn0439-8114.2015.23.051

Analysis of The Optimization of Sulfation Pachymaran Synthesis Technology Process by Response Surface Method and Sulfation Pachymaran Anticoagulant Activity

CHEN Xiao-hong

（College of Life Science，Longyan University，Longyan 364012，F(xiàn)ujian，China）

Abstract：The pachyman was extracted from traditional Chinese medicine Poria cocos，with amino sulfo-nic acid as esterification reagent， treated the pachyman with sulfated processing to make sulfation pachymaran （SP） ?and optimized the conditions of their production processing by response surface method. After purification， structural characterization，sulfation pachymaran（SP） was examined the anticoagulant activity by the anticoagulant effect test in mice. Experimental results showed that the optimal sulfated processing of sulfation pachymaran（SP） were as follows： reaction time 4.0 h，chlorousulfonic acid and pyridine mass ration 3.77∶1，reaction temperature 85 ℃. Under those conditions the degree of substitution was 1.041，the degree of substitution was equal compared with the method of chlorosulfonic acid pyridine in the same conditions. The SP has the characteristics of polysaccharides and sulfate group absorption peak in its ir spectrum after refined， the SP showed high purity and no impurities such as nucleic acids， proteins， and had the characteristics of sulfate group absorption in its UV scanned map. Good anticoagulant activity was showed in the SP， its BT， CT and CT extension rate were significantly improved. The effect on inhibition of thrombin was positively interaction with and SPs concentration， CT and CT extension rate were obviously improved with the do-sage increased by intraperitoneal injection.

Key words： Poria cocos; sulfation; pachymaran（SP）; response surface method; anticoagulant activity

中藥茯苓（Poria cocos）為多孔菌科真菌茯苓的菌核，俗稱云苓、松苓、茯靈，是寄生在松樹根上的菌類植物，形狀像甘薯，外皮黑褐色，里面白色或粉紅色。其原生物為多孔菌科真菌茯苓的干燥菌核，菌核呈球形或不規(guī)則塊狀，大小不一。茯苓味甘、淡，性平，入藥具有利水滲濕、益脾和胃、寧心安神的功用[1]。現(xiàn)代醫(yī)學(xué)研究表明，茯苓多糖為中藥茯苓的主要有效成分，但未經(jīng)結(jié)構(gòu)改性的茯苓多糖對腫瘤抑制率僅0.3%，活性低、水溶性差，難以利用，因此必須對原茯苓多糖進行相應(yīng)的結(jié)構(gòu)改性以提高其活性[2]。……