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-l-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 (P＜0.05).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.%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)时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:采用表达栽体pGEX-6p-1-hri进行GST-hRi的融合表达,并进行纯化;另取犬鼠一次性腹腔注射链脲佐菌素70mg·kg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8 Pg-d)、治疗组(GST-hRJ100 u·d),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,活性为9×10u·mL;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.
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