摘要
为观察金钗石斛生物碱抗糖性白内障作用及其相关蛋白谱。将Wistar大鼠随机分为正常对照组、模型组、样品高、低剂量组和阳性对照组,每组10只,模型组腹腔注射D-半乳糖诱导大鼠白内障模型,样品高、低剂量组和阳性对照组在注射D-半乳糖26 d后形成Ⅱ级白内障后每天分别给予0.36、0.18 g/kg金钗石斛总生物碱灌胃进行实验性治疗,阳性对照组每天以0.22 g/kg石斛夜光丸灌胃,正常对照组每天仅灌胃生理盐水;用裂隙灯观察观察晶状体浑浊度;46 d后处死动物后检测晶状体水溶性蛋白、GSH的含量及T-SOD和MDA的活性;使用双向电泳技术提取分离大鼠晶状体总蛋白及糖基化蛋白,分析比较双向电泳图谱,寻找正常组与模型组、生物碱治疗组大鼠晶状体蛋白质组学差异。结果金钗石斛生物碱高剂量组能明显减轻晶状体混浊度,显著升高晶状体水溶性蛋白、GSH含量及T-SOD活性,降低MDA的活性;2-DE获得了较好的大鼠晶状体总蛋白质及糖基化蛋白质的双向电泳图谱,并观察到模型组有大量蛋白质点表达上调或下调或新蛋白,蛋白质糖基化水平升高,给予高剂量石斛生物碱后,大部分达上调或下调蛋白质恢复到正常的水平,糖基化蛋白降低。表明金钗石斛生物碱具有较好的抗糖性白内障作用,并伴随一系列晶状体蛋白质表达水平的改变,其差异蛋白可能参与了晶状体浑浊过程。
To investigate the mechanisms of the prevention on the cataract in rat lens at the level of proteome by the total alkaloids extracted from Dendrobium nobile Lindl.. Wistar rats were randomly divided into 5 groups （ n = 10） , namely, the normal control, cataract model ,360 mg/kg alkaloids, 180 mg/kg alkaloids, and positive control groups. The cataract model was induced by intragastrical administration of D-galactose into rats. Rats of the alkaloids groups received corresponding amount of alkaloids and those in the positive control group received 220 mg/kg Shihuyeguangwan. The Changes of the lens were examined with slit lamp microscope at defined timegroups received corresponding amount of alkaloids and those in the positive control group received 220 mg/kg Shihuyeguangwan. The changes of the lens were examined with slit lamp microscope at defined time points. The content of water-solubility protein, GSH, and the activity of T-SOD and MDA of each lens was measured with the kits respectively. At the same time, changes in proteomic expression in rats＇ lens were investigated by two-dimensional gel electrophoresis（2-DE）. The lens opacity can be lighten obviously by the total alkaloids, the contents of water-solubility protein, GSH, and the activity of T-SOD in the lens were up-regulated as the activity of MDA was degraded by alkaloids especially the tfigh-dose. The 2-DE maps for proteins of the cataract rats were reported and different proteins were found compared with normal rats. The total alkaloids extracted from Dendrobium nobile Lindl are preventive effect on development of sugar cataract in experimental models. There was a series alteration of protein expression in rats＇ lens. Those proteins maybe involved in the inflammation process of the len.
出处
《天然产物研究与开发》
CAS
CSCD
2008年第4期617-621,共5页
Natural Product Research and Development
基金
广东省自然科学基金项目(06024128)
广东省科技攻关计划项目(2007B020704005)
关键词
金钗石斛生物碱
糖性白内障
双向电泳
晶状体蛋白质
total alkaloids extracted from Dendrobium nobile
diabetic cataract
two-dimensional gel electrophoresis
proteinuria
