【作者】 吴文晔；

【导师】 金坚；

【作者基本信息】 江南大学 ， 微生物与生化药学， 2008， 硕士

【摘要】 糖尿病(diabetes mellitus,DM)是由多种遗传和环境因素相互作用而引起的一组慢性代谢异常综合征。目前常用的口服降糖药有双胍类、磺脲类、噻磋烷二酮类等。这些化学降糖类药物尽管见效较快,但对肝、肾等脏器损害较大,甚至有发生低血糖的危险。一般这类抗高血糖药物仅仅只是降低异常升高的血糖,对于糖尿病患者起到治标不治本的作用。为此,本课题着眼于其病理基础之一“胰岛损伤”,特别是晚期糖尿病,胰岛细胞功能严重衰退,β细胞大量凋亡。针对这一点本课题建立了一个细胞水平上的药物筛选模型,从我国传统中药库中筛选出具有促进β细胞增殖,抑制其凋亡的药物。从而达到治标又治本的目的。且纯天然提取物类药物具有作用温和、毒副作用小等优点,开发利用潜力很大。首先,本论文以大鼠胰岛瘤细胞系RINm5f为基础材料,以GLP-1为阳性参照,用SRB法测定细胞生长曲线及增殖活性。确定了最佳细胞铺板浓度为104个/孔,阳性参照浓度50nmol/L,增殖率最高达30%。确定了中药样品的筛选浓度分别为100mg/L和10mg/L,最佳干预时间72小时,成功建立了体外药物筛选模型。并运用此模型对313味中药进行了初筛和复筛,筛选出蜈蚣、地龙两味中药具有促RINm5f细胞增殖的作用。然后,我们对活性药物进行初步分离。动物类药材蜈蚣,地龙采用水提醇沉方法,然后分别对各分离组分进行细胞活性评价。结果显示只有蜈蚣的50%乙醇醇沉上清组分对RINm5f细胞的生长有显著的促进作用,其他各组分都无显著促增殖活性的作用。最后,本论文选择促细胞增殖率相对较高的中药蜈蚣水提物进行了体内药效的评价。通过比较四氧嘧啶糖尿病小鼠服用蜈蚣水提物前后的生长情况,血糖值,胰岛β细胞修复情况等,初步考察中药蜈蚣水提物的降血糖功效。结果表明:蜈蚣水提物具有良好的体内降血糖功效。更多还原

【Abstract】 Diabetes (diabetes mellitus, DM) is a group of chronic metabolic syndrome arising from the interaction of many genetic and environmental factors. Now commonly used oral hypoglycemic agents are glucophage, glibenclamide and so on. Although these chemical hypoglycemic drugs have fast effects, the liver, kidney and other organs were seriously injuried, or even there is risk of hypoglycemia. Generally such anti-high blood sugar drugs simply reduce the abnormal increased of blood sugar, but they can’t reduce islet damage in patients with diabetes.To avoid such shortage mentioned above, this subject focus on one of the basic pathological event in "islet injury," especially the advanced diabetes, islet severe recession of cell function, large number ofβ-cells apoptosis. In order to figure out this problem, a drug screening model on cell level was established for screening drugs from traditional Chinese medicine with the function of promotingβcell proliferation, and inhibiting apoptosis .The aim is to gain profit from treating the symptoms and cause of disease. And pure extracts of natural drugs play modest role, have slight side effects, and have great potential on being developed and utilized.First, in this experiment, RINm5f cells was basic materials, GLP-1 was positive reference, the activity of cell growth and proliferation determinated by SRB method. The best density of cells determined was 10,000 per hole, the GLP-1 concentration was 50 nmol / L, and the most proliferation rates was at 30%. The best secreen concentrations of Chinese medicine samples were 100 mg / L and 10 mg / L, the best interference time was 72 h. Successfully established a drug screening model in vitro at cell level. And 313 kinds of herbs were secreened by this model, and founed that two Chinese herbal medicines scolopendra and pheretima had the function of promoting proliferation in RINm5f cells.Then, the two drugs which had the function of promoting proliferation in RINm5f cells were preliminary separated by water-alcohol-ways, and evaluated the activity in RINm5f cells. The results showed that only the components dissolved in 50 percents of ethanol of scolopendra had significant effect on promoting proliferation in RINm5f cell, the others had no significant function on promoting proliferation.Finally, the water extracts of scolopendra which had higher rate of promoting the cell proliferation was used to evaluate its activity in vivo. The model of diabetes mellitus was induced by alloxan in mouse. The growth, blood sugar level,β-cell repairing were observed. The results showed that scolopendra had significant effect in decreasing blood glucose in diabetes mellitus mice.更多还原