不同极性石斛生物碱的提取分离及对人晶状体上皮细胞增殖的影响  

Extraction and Isolation of Different Polar Alkaloids from Herba Dendrobii and Their Influences on Human Lens Epithelial Cells Proliferation

作　　者：魏小勇[1] 马伟凤[1] 方花[1] 吴开力[2] 王松[1] 高欣欣[1] 

机构地区：[1]广州中医药大学,广东广州510006 [2]中山大学眼科中心,广东广州510060

出　　处：《广州中医药大学学报》2010年第4期362-366,437,共6页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(编号:30850012);广东省自然科学基金项目(编号:06024128);广东省科技计划项目(编号:2007B020704005;2009B060700070;2009B090300335)

摘　　要：【目的】观察不同极性石斛生物碱对人晶状体上皮细胞(HLEC)的保护作用。【方法】石斛粉末经乙醇提取、降膜浓缩、酸化、盐析、萃取后,水洗、回收提取分离不同极性生物碱。HLEC用含体积分数10%胎牛血清的DMEM培养液培养,石斛生物碱的保护作用实验分为10组:正常对照组,模型组,水溶性生物碱、脂溶性生物碱、低极性生物碱、弱极性生物碱高、低剂量组,剂量均分别为25.0、12.5μg/L;采用四甲基偶氮唑盐(MTT)法检测不同极性生物碱对HLEC增殖的影响。【结果】通过单因素实验,获得了影响不同极性石斛生物碱提取率的关键因素,均为体积分数80%乙醇提取3次,提取3 h时提取率最高;模型组HLEC增殖被H2O2明显抑制,低剂量脂溶性生物碱组细胞抑制率显著降低,与模型组比较差异有显著性意义(P<0.05)。【结论】较低剂量的脂溶性生物碱能更好地抑制H2O2对晶状体上皮细胞的氧化损伤。Objective To observe the protection of human lens epithelial cells（HLEC） by different polar alkaloids extracted from Herba Dendrobii（HD）.Methods We extacted the Herba Dendrobii powder with ethanol,and then treated the extract with falling-film concentration,acidification,salting out,chloroform extraction,and washing with water.Different polar alkaloids were extracted from HD after the above treatment.The protective effect of HD alkaloids was observed on HLEC,which were cultured with DMEM medium containing 10% fetal calf serum.Ten groups were set up for the experiment： normal control group,model group,high-and low-dose water-soluble alkaloids groups,high-and low-dose fat-soluble alkaloids groups,high-and low-dose low-polar alkaloids group,and high-and low-dose weak-polar alkaloids groups.The high-dose dosage of the alkaloids was 25.0 μg/L and low-dose dosage was 12.5 μg/L.Methyl thiazolyl tetrazolium（MTT） assay was used to evaluate the proliferation of HLEC under the different conditions of interventions.Results The single-factor experiments showed that the highest extracting rate of HD alkaloids was obtained under the conditions of extracting the powder with 80% ethanol for 3 times and for 3 hours.The results of protective experiment showed that the proliferation of HLEC in the model group was inhibited by hydrogen peroxide（H2O2）,and the inhibitive rate was lower in low-dose fat-soluble alkaloids group than that in the model group（P〈0.05）.Conclusion Low-dose fat-soluble HD alkaloids have a better effect on inhibiting the H2O2-induced oxidative injury of HLEC.

关 键 词：石斛/分离与提纯 石斛/药理学 晶状体上皮细胞/病理学 细胞培养 

分 类 号：R285.5[医药卫生—中药学]