糖基化终末产物及其受体轴体系在糖尿病性白内障中的作用与机制

发布时间：2018-06-07 08:18

  本文选题：糖尿病性白内障 + 糖基化终末产物　； 参考：《泸州医学院》2014年硕士论文

【摘要】：目的：建立糖尿病大鼠模型，动态观察晶状体的混浊程度、氧化损伤程度、晶状体内糖基化终末产物（AGEs）浓度、糖基化终末产物受体（RAGE）及核因子-κBp65（NF-κBp65）mRNA和蛋白水平的变化，探讨AGEs-RAGE轴体系在糖尿病性白内障发生发展中的作用及机制。方法：随机将无眼部疾病的SD大鼠分成正常组（32只）和糖尿病组（36只），糖尿病组以单次腹腔注射1%链脲佐菌素（STZ）溶液65mg/kg诱导糖尿病，正常组注射同等体积的0.1mmol/L柠檬酸盐缓冲液。3天后空腹（禁食12小时）尾静脉血糖高于16.6mmol/L为糖尿病成模。每天观察大鼠一般情况，每两周检测记录血糖、体质量，每周麻醉及复方托吡卡胺滴眼液散瞳后在裂隙灯下观察晶状体混浊情况。在实验4、8、12周时分批处死大鼠，即刻取出双眼晶状体。每组各取3枚晶状体制成石蜡切片，HE染色观察晶状体细胞学及免疫组化检测晶状体上皮细胞中RAGE、NF-κBp65表达；5枚晶状体提取RNA后RT-PCR检测RAGE mRNA和NF-κBp65mRNA表达；12枚晶状体匀浆后检测AGEs浓度、T-SOD、GSH-PX和CAT活力及MDA含量。结果：1.注射STZ后大鼠出现糖尿病典型症状，血糖明显高于正常组（t4周=22.84，P=0.000；t8周=26.01，P=0.000；t12周=30.94，P=0.000）。早期糖尿病大鼠多食但体质量增加不明显，随着病程进展逐渐消瘦，这与正常组的体质量逐渐增加有显著差别（t4周=6.54，P=0.000；t8周=20.39，P=0.000；t12周=46.91，P=0.000）。2.正常组大鼠晶状体始终透明；糖尿病组在实验4周时部分晶状体周边部及前囊膜下出现细小空泡，随后空泡向中心扩展，瞳孔区可见团块样混浊，8周时有皮质混浊，此后混浊逐渐累及核周皮质及核，至12周时出现晶状体完全混浊。3. HE染色显示糖尿病大鼠晶状体上皮细胞（LECs）形态多样，，胞质淡染伴有空泡、胞核缩小；晶状体纤维排列紊乱，其间有较多囊泡及水裂。4.正常组LECs中几乎无RAGE表达、NF-κBp65在胞质低表达，糖尿病组RAGE和NF-κBp65均呈强阳性表达（RAGE：t4周=90.73，P=0.000；t8周=176.77，P=0.000；t12周=148.67，P=0.000；NF-κBp65：t4周=83.98，P=0.000；t8周=130.61，P=0.000；t12周=195.69，P=0.000），且随着病程发展表达进行性增加。糖尿病组晶状体中RAGEmRNA和NF-κBp65mRNA的表达均明显高于正常组（RAGE：t4周=20.36，P=0.000；t8周=29.90，P=0.000；t12周=57.19，P=0.000；NF-κBp65：t4周=11.34，P=0.000；t8周=32.21，P=0.000；t12周=41.03，P=0.000）。5.在实验4、8、12周时糖尿病组晶状体中AGEs浓度均高于正常组（t=9.01，P=0.000；t=6.45，P=0.000；t=9.38，P=0.000），且随着病程延长出现AGEs大量蓄积。6.糖尿病组晶状体中SOD、GSH-PX、CAT的活力均明显降低正常组，MDA含量显著升高（P均小于0.05）。结论：1.糖尿病大鼠晶状体内AGEs蓄积并与RAGE结合可增强NF-κB活性；活化的NF-κBp65可上调RAGE的转录与合成，并与更多的AGEs作用。2.随着病程延长，糖尿病大鼠晶状体内生成AGEs增多，抗氧化酶活性降低，晶状体上皮细胞氧化损伤加重，晶状体混浊加重。3.晶状体内AGEs-RAGE轴体系介导的氧化应激、非酶糖基化作用中多种信号转导通路并形成级联放大反应可能是糖尿病性白内障发生发展的机制之一，NF-κB是此过程中关键的中间物。
[Abstract]:Objective : To establish a model of diabetic rats , to dynamically observe the degree of opacity , oxidative damage , end - of - end product ( AGE ) , end - product receptor ( RAGE ) and nuclear factor - 魏B ( NF - 魏B ) mRNA and protein levels in diabetic rats .
The expression of RAGE mRNA and NF - 魏B p65mRNA was detected by RT - PCR after extraction of RNA from five lenses .
Results : 1 . The diabetic symptoms of diabetic rats were higher than those in the normal group ( t4 weeks = 22.84 , P = 0.000 ) .
8 weeks = 26.01 , P = 0.000 ;
12 weeks = 30.94 , P = 0.000 ) . In the early diabetic rats , the body mass of the rats was not significantly increased , and the body mass of the normal group increased gradually with the progressive loss of the disease course ( t4 = 6.54 , P = 0.000 ) .
8 weeks = 20.39 , P = 0.000 ;
12 weeks = 46.91 , P = 0.000 ) .
In the diabetic group , small vacuoles appeared in the peripheral part of the lens and the anterior capsule at 4 weeks of the experiment , followed by the extension of the cavity to the center . The area of the pupil was turbid , and the cortex was turbid at 8 weeks . After that , the opacity gradually increased to the cortex and the nucleus , and the lens was completely cloudy at 12 weeks . HE staining showed that the lens epithelial cells ( LECs ) of diabetic rats were varied in shape , and the cytoplasm was light - stained with vacuoles and the nuclei were reduced ;
4 . There was almost no RAGE expression in LECs in the normal group , the low expression of NF - NF - 魏B in the normal group , the low expression of NF - 魏B , the expression of RAGE and NF - 魏B in the diabetic group ( RAGE : t4 = 90.73 , P = 0.000 ) .
8 weeks = 176.77 , P = 0.000 ;
12 weeks = 148.67 , P = 0.000 ;
NF - 魏B P65 : t4 week = 83.98 , P = 0.000 ;
8 weeks = 130.61 , P = 0.000 ;
The expression of RAGEmRNA and NF - 魏B p65mRNA in diabetic group was significantly higher than that in the normal group ( RAGE : t4 = 20.36 , P = 0.000 ) .
8 weeks = 29.90 , P = 0.000 ;
12 weeks = 57.19 , P = 0.000 ;
NF - 魏B P65 : t4 week = 11.34 , P = 0.000 ;
8 weeks = 32.21 , P = 0.000 ;
At the 4th , 8th and 12th week of the experiment , the concentration in the lens of the diabetic group was higher than that in the normal group ( t = 9.01 , P = 0.000 ) .
t=6.45,P=0.000锛

本文编号：1990503