地黄多糖的提取纯化及其对糖尿病小鼠血糖的影响研究

Extraction and purifction of rehmannia glutinosa polysaccharide and its effect on blood glucose in diabetic mice

目的研究从地黄中分离纯化多糖的方法,并探讨地黄多糖对四氧嘧啶诱导的糖尿病小鼠的降血糖作用。方法地黄粉末石油醚、乙醚除去脂溶性杂质,80%的乙醇脱单糖、低聚糖,热水浸提液经浓缩,醇析,Savagc法除蛋白,离子水逆流透析,H2O2脱色得地黄粗多糖。苯酚-浓硫酸比色法测多糖含量。将腹腔注射四氧嘧啶建立的糖尿病小鼠动物模型动物随机分为模型组、地黄多糖治疗组(RGP治疗组)、阿卡波糖治疗组(AR治疗组),另设立正常对照组。每组给药后4周末测定空腹12h血糖(GLU)、甘油三酯(TG)、极低密度脂蛋白-胆固醇(VLDL-C)、总胆固醇(TC)的含量及小鼠体重,对各组的血糖和血脂水平进行比较分析。结果①490nm为样品的吸收波长,线形回归方程为:Y=0.0056X+0.4361,相关系数R2=0.9995;地黄粗多糖的提取率为45.5%,总糖含量为35.2%。②与正常组相比,模型组体重降低、血糖升高,相比较有显著性差异(P<0.05);AR治疗组、RGP治疗组与模型组相比,体重增加、血糖降低,相比较有显著性差异(P<0.05)。③模型组血清血脂水平显著高于正常组(P<0.05);AR治疗组、RGP治疗组与模型组比较,TG、VLDL-C降低,相比较有显著性差异(P<0.05);治疗组与模型组比较TC水平变化不大,差异无统计学意义(P>0.05)。结论该方法简便可行,可以作为地黄多糖提取分离纯化的有效方法;地黄多糖具有能缓解糖尿病小鼠体重降低,降低糖尿病小鼠血糖水平,调节血脂紊乱的作用。

OBJECTIVE To study the extraction and purifction of rehmannia glutinosa polysaccharide（RGP）and its effect on blood glucose in diabetic mice induced by the alloxan.METHODS The rehmannia glutinosa powder was removed the fat-soluble impurities by Petroleum ether and ethyl ether,the monosaccharides and oligosaccharides was peeled off using 80% ethanol.The rehmannia glutinosa polysaccharide was extracted and purified through hot water extracted and concentrated,precipitated with ethanol,peeled off protein with Savagc method,deionized water counter current dialysis,removed the pigment in H2O2.The RGP content was detected with Phenol-concentrated sulfuric acid colorimetry.Alloxan was administered intraperitoneally to establish diabetic models in mice,after the success of modeling,experimental animals were divided into the model group,the RGP treatment group,AR treatment group and set up a normal control group.4 weeks after treatment,the 12h fasting blood glucose（GLU）,triglyceride（TG）,very low density lipoprotein cholesterol（VLDL-C）,total cholesterol（TC）levels and body weight of mice were detected,the each group＇s blood glucose and lipid levels were ana-lyzed.RESULTS ①The samples＇ UV absorption was 490nm,linear regression equation：Y=0.0056X＋0.4361,correlation coefficient R2=0.9995;the extraction rate of RGP was 45.5%,total sugar content was 35.2%.②Compared with normal group,the weight loss and elevated blood glucose in the model group,the difference was significant（P0.05）;compared with model group,the gain weight and decreased blood glucose in the AR treatment group and RGP treatment group,the difference was significant（P0.05）.③The serum lipid levels of the model group were significantly higher than that of the normal group（P0.05）;Compared with the model group,the TG and VLDL-C levels of the AR treatment group and RGP treatment group decreased,the difference was significant（P0.05）;compared with model group,the TC levels of treatment group was no significantly difference（P0.05）.CONCLUSION This method is simple and feasible,can be used as a effective methods of RGP extraction and purification;RGP can alleviate the weight loss of diabetic mice,reduce blood glucose levels in diabetic mice and regulated blood lipid disorder.