3-氨基苯甲酰胺抑制STZ诱导的糖尿病大鼠晶状体混浊机制的初步研究

发布时间：2018-07-02 08:45

本文选题：聚ADP核糖聚合酶 + 3-氨基苯甲酰胺　；参考：《泸州医学院》2012年硕士论文

【摘要】：目的：本实验主要通过聚ADP核糖聚合酶(poly ADP-ribose polymerase,PARP)抑制剂3-氨基苯甲酰胺(3-aminobenzene,3-AB)对链脲佐菌素（streptozotocin，STZ）诱导的糖尿病大鼠进行干预，，观察其晶状体混浊程度的进展以及对相关指标的检测，探讨PARP抑制剂对延缓糖尿病大鼠晶状体混浊的机制。方法： 1、体重200±20g雄性Wistar大鼠40只，进行裂隙灯检查排除晶状体病变。适应性喂养3d后自由饮水，禁食24h，随机抽取9只作为正常对照组(A组)，腹腔注射0.1mol/L柠檬酸盐缓冲液（50mg/kg，pH4.5）,其余大鼠则予以腹腔注射1%链脲佐菌素(Stretozocin, STZ)柠檬酸缓冲液（0.1mol/L柠檬酸盐缓冲液临时配制，pH4.5）。3d后尾静脉取血，测得空腹血糖≥16.7mmol/L为糖尿病大鼠。 2、将糖尿病大鼠随机分为糖尿病组（B组）和3-氨基苯甲酰胺干预组（C组）。自建立模型后第三天起，C组每日给予3-AB30mg/kg灌胃，A组和B组每天给予等体积0.9%N.S灌胃。实验中观察记录大鼠体重等一般情况，每周检测一次空腹血糖。 3、1%复方托品酰胺眼液充分散瞳后在裂隙灯下观察并记录各组晶状体混浊进展情况，每周一次，记录参照国外医学眼科学分册的晶状体混浊分级方法。分别于给药后2w、4w、8w分批处死各组大鼠。每组各取3只眼球固定包埋后，做石蜡切片用于免疫组化SP法检测晶状体上皮细胞中基质金属蛋白-2(matrix metalloproteinase-2,MMP-2)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的表达情况；另外3只分离晶状体后分别检测谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-PX）和超氧化物歧化酶（superoxide dismutase,SOD）的活性以及丙二醛（maleicdialdehyde,MDA）和糖基化终末产物（advanced glycation end products,AGE）的含量。结果： 1、选取9只作为正常对照组(A组)大鼠的生长状况良好,采用STZ制作糖尿病大鼠模型,最终18只纳入实验,均出现三多一少的典型糖尿病表现。 2、在实验2、4、8周，B、C组血糖高于A组，差异具有统计学意义(P＜0.01)，但B组与C组相比，血糖无差异(P＞0.05)。 3、在整个实验过程中，A组始终保持晶状体透明，而随着实验的进行，3周时B组、C组均开始出现不同程度的晶状体混浊。4周和8周时A、B、C三组间晶状体混浊程度有明显差异(P＜0.01)，且4周和8周时B组晶状体混浊程度比A组重(P＜0.01)。 4、免疫组化结果：MMP-2在A组几乎没有表达，而随着实验的进行B、C组表达逐渐增强，且B组强于C组(P＜0.01)；bFGF在A、B、C组则均呈阳性表达，A组与B、C组之间表达有明显差异(P＜0.01)，且B、C组随着实验的进行而增强，但B、C组之间bFGF的表达无差异(P＞0.05)。 5、氧化指标的检测：A组GSH-px和SOD的活性均高于B、C两组(P＜0.01)，但在各时间段B组均低于C组(P＜0.01, P＜0.05)；MDA的含量在B、C组增加，与A组比较(P＜0.01)，且B组高于C组(P＜0.05)。 6、B、C组在各时间段AGE的含量均高于A组；在2周、4周时B组高于C组(P＜0.05)，但在8周时无差异(P＞0.05)。结论： 1、PARP抑制剂3-AB对糖尿病大鼠晶状体具有一定保护作用，可以延缓糖尿病性白内障的发生发展。 2、PARP抑制剂3-AB通过减轻高糖所致的晶状体上皮细胞氧化损伤和非酶糖基化水平而抑制晶状体的混浊。 3、PARP抑制剂3-AB抑制MMP-2的表达减轻晶状体上皮细胞外基质降解，从而达到延缓糖尿病性白内障的作用。 4、bFGF参与了糖尿病性白内障的发生发展。
[Abstract]:Objective:
In this experiment, ADP ribose polymerase (poly ADP-ribose polymerase, PARP) inhibitor, 3- amino benzoformamide (3-aminobenzene, 3-AB), was used to interfere with streptozotocin (streptozotocin, STZ) induced diabetic rats. The progress of the degree of lens opacification and the detection of the related indexes were observed. The delay of the inhibitor was discussed. The mechanism of lens opacification in diabetic rats.
Method:
1, 40 male Wistar rats with weight of 200 20g were checked out of lens lesions by slit lamp examination. After feeding 3D free drinking water and fasting 24h, 9 rats were randomly selected as normal control group (A group) with 0.1mol/L citrate buffer (50mg/kg, pH4.5) intraperitoneally, and the rest rats were intraperitoneally injected with 1% streptozotocin (Stretozocin, STZ) lime (Stretozocin, STZ). Blood glucose was obtained from tail vein after citric acid buffer solution (0.1mol/L citrate buffer was temporarily formulated, pH4.5).3d, and fasting blood glucose (16.7mmol/L) was measured in diabetic rats.
2, diabetic rats were randomly divided into diabetes group (group B) and 3- amino benzamide intervention group (group C). From the third day after the establishment of the model, group C was given 3-AB30mg/kg gavage every day, and group A and B group were given equal volume of 0.9%N.S every day. The experiment observed the body weight of rats and other cases, and tested the fasting blood glucose once a week.
3,1% compound entrapment amide eye solution was fully scattered under the slit lamp, observed and recorded the progress of lens opacities in each group. Once a week, the lens turbidity classification method was recorded with reference to foreign medical ophthalmology division. The rats were killed in groups of 2W, 4W, and 8W after administration. Each group took 3 eyeballs and embedded to make paraffin slices. The expression of matrix metalloprotein -2 (matrix metalloproteinase-2, MMP-2) and basic fibroblast growth factor (basic fibroblast growth factor, bFGF) in lens epithelial cells was detected by immuno histochemical SP, and the other 3 separate lenses were used to detect the cereal caspase peroxidase (glutathione peroxidase,) and ultra The activity of oxide dismutase (superoxide dismutase, SOD) and the content of malondialdehyde (maleicdialdehyde, MDA) and the end products of glycosylation (advanced glycation end products, AGE).
Result:
1, the growth status of 9 rats in the normal control group (group A) was good, and the diabetic rat model was made with STZ, and the final 18 were included in the experiment.
2, at the experimental 2,4,8 weeks, the blood glucose in group B and C was higher than that in group A, the difference was statistically significant (P < 0.01), but there was no difference in blood glucose between B group and C group (P > 0.05).
3, during the whole experiment, the A group always kept the lens transparent, and with the experiment, the B group in the group of B and the group C began to appear different degree of lens opacities at the time of 3 weeks and the 8 weeks of A, B, and C three were significantly different (P < 0.01), and the degree of turbidity in the B group was heavier than the A group (P < 0.01) at the 4 and 8 weeks.
4, the results of immunohistochemistry: MMP-2 was almost not expressed in group A, but with the experiment of B, the expression of C group was increased gradually, and B group was stronger than group C (P < 0.01); bFGF in A, B, C group was positive, and there was a significant difference between A group and C group (0.01). P > 0.05).
5, the detection of oxidation index: the activity of GSH-px and SOD in group A was higher than that of B, C two (P < 0.01), but in each time period B group was lower than that of C group (P < 0.01, P < 0.05), MDA content was increased in B, and compared with that of the group (0.01), and the group was higher than that of the group (0.05).
6, the content of AGE in B and C groups was higher than that in group A at each time interval. At 2 weeks and 4 weeks, the B group was higher than that of C group (P < 0.05), but there was no difference at 8 weeks (P > 0.05).
Conclusion:
1, PARP inhibitor 3-AB has a certain protective effect on the lens of diabetic rats, and it can delay the occurrence and development of diabetic cataract.
2, PARP inhibitor 3-AB inhibits lens opacity by reducing the oxidative damage and non enzymatic glycosylation level of lens epithelial cells induced by high glucose.
3, PARP inhibitor 3-AB inhibits the expression of MMP-2 and alleviates the degradation of extracellular matrix in lens epithelial cells, thereby slowing the progression of diabetic cataract.
4, bFGF is involved in the occurrence and development of diabetic cataract.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R776.1

【共引文献】