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融合蛋白GST-hRI对糖尿病性白内障模型大鼠晶状体浑浊程度的改善作用研究
引用本文:刘芳君,郝海峰,王冬梅,杨帆,田余祥,赵宝昌,李晶敏.融合蛋白GST-hRI对糖尿病性白内障模型大鼠晶状体浑浊程度的改善作用研究[J].中国药房,2011(5):395-398.
作者姓名:刘芳君  郝海峰  王冬梅  杨帆  田余祥  赵宝昌  李晶敏
作者单位:大连医科大学生物化学教研室;河南省卫生学校生物化学教研室;大连医科大学机能实验室;大连医科大学附属第二医院眼科教研室;
基金项目:辽宁省教育厅科学技术研究项目(2008171)
摘    要:目的:研究融合蛋白谷胱甘肽硫转移酶-人核糖核酸酶抑制因子(GST-hRI)对糖尿病性白内障模型大鼠晶状体浑浊程度的改善作用。方法:采用表达载体pGEX-6p-1-hri进行GST-hRI的融合表达,并进行纯化;另取大鼠一次性腹腔注射链脲佐菌素70mg·kg-1建立糖尿病性白内障模型,随机分为正常对照组、模型组(生理盐水)、白内停组(白内停8μg·d-1)、治疗组(GST-hRI100u·d-1),每组12只,双侧点眼给予相应药物,每天4次,连续6周,每周裂隙灯下观察大鼠晶状体浑浊程度,6周后处死大鼠,考察各组大鼠晶状体中超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、谷胱甘肽过氧物酶(GSH-Px)及丙二醛(MDA)含量变化。结果:成功诱导表达并纯化GST-hRI,蛋白含量为0.413mg·mL-1,活性为9×103u·mL-1;与模型组和白内停组比较,治疗组大鼠晶状体浑浊程度显著减轻,其SOD、GSH、GSH-Px含量明显增加,MDA含量明显下降(P<0.05)。结论:融合蛋白GST-hRI具有明显的抗氧化作用,能够预防或延缓糖尿病性白内障晶状体浑浊。

关 键 词:融合蛋白  谷胱甘肽硫转移酶-人核糖核酸酶抑制因子  糖尿病性白内障  大鼠  改善

Improvement Effect of Fusion Protein GST-hRI on Lens Turbidness in Rats with Diabetic Cataract
LIU Fang-jun,HAO Hai-feng,YANG Fan,TIAN Yu-xiang,ZHAO Bao-chang LIU Fang-jun,HAO Hai-feng WANG Dong-mei LI Jing-min.Improvement Effect of Fusion Protein GST-hRI on Lens Turbidness in Rats with Diabetic Cataract[J].China Pharmacy,2011(5):395-398.
Authors:LIU Fang-jun  HAO Hai-feng  YANG Fan  TIAN Yu-xiang  ZHAO Bao-chang LIU Fang-jun  HAO Hai-feng WANG Dong-mei LI Jing-min
Institution:LIU Fang-jun, HAO Hai-feng, YANG Fan, TIAN Yu-xiang, ZHAO Bao-chang(Dept. of Biochemistry, Dalian Medical University, Dalian 116044, China) LIU Fang-jun,HAO Hai-feng(Dept. of Biochemistry,Health School of He’nan Province,Anyang 455000,China) WANG Dong-mei(Function Laboratory of Dalian Medical University, Dalian 116044, China) LI Jing-min(Dept. of Ophthalmology, The Second Affiliated Hospital of Dalian Medical University, Dalian 116023, China)
Abstract:OBJECTIVE:To observe the effect of fusion protein glutathione sulfurtransferase-human ribonuclease inhibitor(GST-hRI)on lens turbidness in rats with diabetic cataract . METHODS:The pGEX-6p-1-hri expressive plasmid was induced to express GST-hRI, and the expressed GST-hRI was purified. Rats were given intraperitoneal injection of streptozotocin 70 mg·kg-1 to establish diabetic cataract model. Except normal control group, model rats were randomly divided into model group (normal saline), pirfenoxone group (pirfenoxone 8 μg·d-1) and treatment group (GST-hRI 100 u·d-1) with each group of 12 rats. Each group was given relevant drugs by eye-drop application 4 times a day for 6 weeks. The lens turbidness degree was inspected with the slit lamp once a week. The rats were sacrificed on the sixth week and their lens were removed to determine the levels of superoxidase dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA). RESULTS:The expression of GST-hRI had been induced and purified successfully and the content of protein was 0.413 mg·mL-1 and activity was 9×103 u·mL-1. Compared with model group and pirfenoxone group, the degree of lens turbidness in treatment group was significantly relieved. The content of SOD, GSH and GSH-Px in treatment group increased and the content of MDA decreased significantly (P0.05). CONCLUSION:The fusion protein GST-hRI has obvious antioxidant effect and has significant prevention and treatment effect on lens turbidness in rats with diabetic cataract.
Keywords:Fusion protein  Glutathione sulfurtransferase-human ribonuclease inhibitor  Diabetic cataract  Rats  Improvement  
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