PPAR-γ激动剂对早期糖尿病大鼠视网膜的保护作用

发布时间：2018-04-24 06:32

本文选题：糖尿病性视网膜病变 + 过氧化物酶体增生物激活受体-γ　；参考：《泸州医学院》2010年硕士论文

【摘要】：目的：应用过氧化物酶体增生物激活受体-y(peroxisome proliferator-activated receptor-gamma, PPAR-y)激动剂罗格列酮对早期糖尿病(diabetes mellitus, DM)大鼠模型进行干预,观察PPAR-y激动剂对DM大鼠视网膜中基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)、基质金属蛋白酶-2 (matrix metalloproteinases-2, MMP-2)的表达及血-视网膜屏障(blood-retina barrier, BRB)通透性的影响,进而从分子学水平上阐明PPAR-y激动剂对早期糖尿病性视网膜病变(diabetic retinopathy, DR)的保护作用,为预防及早期治疗DR提供一种新思路。方法：选择114只健康成年雄性Wistar大鼠,随机分为正常对照组(C组,30只),糖尿病组(DM组,42只),DM+罗格列酮组(DM+R组,42只)。予以DM组及DM+R组链脲佐菌素(Streptozocin, STZ)按50mg/kg一次性腹腔注射建立DM动物模型。将死亡及未成模大鼠剔除出实验组,按实验时间点,将以上三组大鼠进一步分为C 4w组(10只)、8w组(10只)、12w组(10只),DM 4w组(10只)、8w组(10只)、12w组(10只)及DM+R 4w组(10只)、8w组(10只)、12w组(10只)。自DM模型建立后第三天起,DM+R 4w、8w及12w组每日给予罗格列酮3mg/kg灌胃,每周测血糖和体重两次。分别于给药后4w、8w及12w分别处死各组大鼠,每组各取5只大鼠眼球行BRB通透性测定；另5只大鼠摘除其左眼球行视网膜消化铺片观察视网膜微血管变化情况,计数内皮细胞(endothelial cell, E)、周细胞(perithelial cell, P),计算E/P值；摘除其右眼球行石蜡包埋,应用免疫组化方法检测视网膜中MMP-9及MMP-2的表达情况,并对实验结果进行统计学分析。结果：(1)BRB通透性测定结果显示：在各实验时间点,DM及DM+R组BRB通透性较相应C组均增加(P0.01)；DM+R组较相应DM组BRB通透性明显降低(P0.01),且呈时间依赖性改变；但DM+R组较各相应时间点C组BRB通透性增加(P0.01)。(2)视网膜消化铺片结果显示：DM 4w组与C组及DM+R 4w组比较,所有大鼠视网膜毛细血管走行、血管壁细胞数目及血管形态均无明显异常改(P0.05)；DM 8w及12w组与相应C 8w及12w组比较,毛细血管迂曲,管径粗细不一,血管壁P数量明显减少(P0.01),E数量增多(P0.01)；在8w及12w时,DM+R组与相应的DM组比较,可见血管壁P丢失数量减少(P0.01),E增生现象减轻(P0.01),且呈时间依赖性改变；(3)免疫组化检测结果显示：DM 4w、8w及12w组较相应C组MMP-2及MMP-9表达增多(P0.01)；DM+R各组与相应各时间点的C组比较,MMP-2及MMP-9表达均增多(P0.01),但与相应各时间点的DM组比较,MMP-2及MMP-9表达均减少(P0.01)。组内比较见C 4w、8w及12w组视网膜中MMP-2阳性表达细胞无明显差异(P0.05)；DM 8w组视网膜中MMP-2及MMP-9阳性表达细胞明显多于DM 4w组(P0.01)；DM 12w组视网膜中MMP-2及MMP-9阳性表达细胞明显多于DM 4w、8w组(P0.01)；DM+R 8w、12w组视网膜中MMP-2及MMP-9阳性表达细胞明显多于DM+R 4w组(P0.01),但DM+R 8w与12w组间比较差异无统计学意义(P0.05)。结论：(1)采用STZ一次性腹腔注射建立的DM大鼠模型可用于早期DR的相关研究。(2)MMP-2及MMP-9在早期DR的发病过程中发挥着重要作用。(3)PPAR-γ激动剂罗格列酮可以抑制早期DM大鼠视网膜中MMP-2及MMP-9的表达,降低BRB通透性,对早期DM大鼠视网膜具有一定的保护作用。
[Abstract]:Objective: to use the peroxisome activation receptor -y (peroxisome proliferator-activated receptor-gamma (PPAR-y) agonist, rosiglitazone, to intervene in the rat model of diabetes mellitus (DM), and to observe the matrix metalloproteinase -9 (matrix) in the retinal membrane of DM rats. P-9), the expression of matrix metalloproteinase -2 (matrix metalloproteinases-2, MMP-2) and the influence of the permeability of the blood retinal barrier (blood-retina barrier, BRB), and then elucidate the protective effect of PPAR-y agonists on the early diabetic retinopathy (diabetic retinopathy, DR) from the molecular level, for prevention and early treatment. Method: 114 healthy adult male Wistar rats were selected and randomly divided into normal control group (group C, 30), diabetes group (group DM, 42), DM+ rosiglitazone group (DM+R group, 42 rats). DM group and DM+R group of streptozotocin (Streptozocin, STZ) were injected into DM animal model according to 50mg/kg. Death and unformed models were established. Rats were removed from the experimental group, and the three groups were further divided into C 4W group (10 rats), group 8W (10), group 12W (10), group DM 4W (10), 8W group (10), 12W group (10) and DM+R 4W group (10), 8W group (10), 10 only. Gastric blood glucose and weight were measured two times a week. The rats were killed in each group after 4W, 8W and 12W respectively after administration, respectively, and 5 rats in each group were measured for BRB permeability. The other 5 rats were removed from the left eyeball to observe retinal microvascular changes, counting the endothelial cells (endothelial cell, E), and pericytes (perithelial cell). P), the value of E/P was calculated, paraffin embedded in the right eye was removed and the expression of MMP-9 and MMP-2 in the retina was detected by immunohistochemical method, and the results of the experiment were statistically analyzed. Results: (1) the results of BRB permeability test showed that the BRB permeability of DM and DM +R groups increased (P0.01) in each experiment time point, and the DM+R group was more than that of the DM+R group. The BRB permeability of the corresponding DM group was significantly decreased (P0.01), and the BRB permeability of group DM+R increased (P0.01) compared with the C group at the corresponding time point (P0.01). (2) the results of retinal digestion showed that there was no obvious difference between the DM 4W group and C group and DM+R 4W group, the capillary blood tube of the retina of all rats, the number of vascular wall cells and the morphology of blood vessels were not obvious. Compared with the corresponding C 8W and 12W group, the DM 8W and 12W group were compared with the corresponding C 8W and 12W group. The capillary was circuitous, the diameter of the tube was not one, the number of P in the vessel wall decreased significantly (P0.01), and the number of E increased (P0.01). (3) the results of immunohistochemical detection showed that the expression of MMP-2 and MMP-9 in the group of DM 4W, 8W and 12W increased more than that of the corresponding C group (P0.01), and the MMP-2 and MMP-9 expressions were increased compared with those of the corresponding C groups at the corresponding time points. There was no significant difference in the positive expression of MMP-2 in the membrane (P0.05), and the positive expression of MMP-2 and MMP-9 in the retina of the DM 8W group was more than that of the DM 4W group (P0.01), and the cells in the retina of the retina of the DM 12W group were more than those in the retina, and the positive cells in retina and the retina were more than those in the group of DM 4W. (P0.01), but there is no significant difference between DM+R 8W and 12W group (P0.05). Conclusion: (1) the DM rat model established by one-off intraperitoneal injection of STZ can be used for early DR related studies. (2) MMP-2 and MMP-9 play an important role in the early DR pathogenesis. (3) the PPAR- gamma agonist rosiglitazone can inhibit the early retina of rat retina The expression of MMP-2 and MMP-9 decreased the permeability of BRB and protected the retina of early DM rats.

【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1

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