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目的观察PPAR-γ激动剂15d-PGJ2对糖尿病脑缺血再灌注大鼠脑缺血再灌注损伤小胶质细胞活化及神经细胞凋亡的影响。方法成年SD大鼠80只,随机分为4组:(1)假手术组;(2)正常血糖脑缺血组;(3)糖尿病脑缺血组;(4)糖尿病脑缺血+15d-PGJ2干预组。采用链脲佐菌素诱导糖尿病,应用改良的Zea-Longa法制作大鼠大脑中动脉闭塞再灌注模型。糖尿病脑缺血组+15d-PGJ2干预组在成功制备糖尿病大鼠模型后给予15d-PGJ2 200μg·kg~(-1)·d~(-1)腹腔注射21 d后应用改良的Zea-Longa法制作大鼠大脑中动脉闭塞再灌注模型,再灌注后3 h腹腔注射15d-PGJ2 400μg·kg~(-1),以后6 d每天给予15d-PGJ2 200μg·kg~(-1)·d~(-1)腹腔注射。每组分别于24 h、7 d各处死一批大鼠,并随机分为2组:一组行免疫组化法检测小胶质细胞CD68的表达水平及ELISA检测TNF-α与IL-1β水平,另一组用TUNEL法原位标记DNA片段检测凋亡细胞计数。结果正常血糖脑缺血组、糖尿病脑缺血组、糖尿病脑缺血+15d-PGJ2干预组与假手术组比较,再灌注24 h、再灌注7 d CD68阳性面积、TNF-α与IL-1β水平、神经细胞凋亡率均明显增加(P<0.05);糖尿病脑缺血组在再灌注24 h、再灌注7 d CD68阳性面积、TNF-α与IL-1β水平、神经细胞凋亡率明显高于正常血糖脑缺血组(P<0.05);糖尿病脑缺血+15d-PGJ2干预组再灌注24 h、再灌注7 d CD68阳性面积、TNF-α与IL-1β水平、神经细胞凋亡率低于未干预组(P<0.05)。结论糖尿病脑缺血组与正常血糖脑缺血组相比较CD68阳性面积更大、TNF-α与IL-1β水平更高及神经细胞凋亡率更高;15d-PGJ2可减少糖尿病脑缺血大鼠小胶质细胞激活、减少炎症因子分泌、降低神经细胞凋亡率。
Objective To observe the effect of PPAR-γ agonist 15d-PGJ2 on microglial activation and neuronal apoptosis during cerebral ischemia-reperfusion injury in diabetic rats. Methods Eighty adult SD rats were randomly divided into 4 groups: (1) sham operation group, (2) normal blood-glucose-induced cerebral ischemia group, (3) diabetic cerebral ischemia group, (4) diabetic cerebral ischemia + 15d- PGJ2 intervention group. Diabetes was induced by streptozotocin (STZ) .According to Zea-Longa method, middle cerebral artery occlusion-reperfusion model was established. Diabetic cerebral ischemic group + 15d-PGJ2 intervention group was given diabetic rats model successfully after giving 15d-PGJ2 200μg · kg -1 d -1 for 21 d, and then the modified Zea-Longa method The model of middle cerebral artery occlusion and reperfusion was established in rats. The rats were injected intraperitoneally with 15d-PGJ2 400μg · kg ~ (-1) 3h after reperfusion and then treated with 15d-PGJ2 200μg · kg ~ (-1) d ~ -1) intraperitoneal injection. Rats in each group were killed at 24 h and 7 d, and randomly divided into two groups: one group were immunohistochemically detected CD68 expression of microglia and the level of TNF-α and IL-1β by ELISA , And the other group detected the apoptotic cell counts by DNA labeling in situ using TUNEL method. Results The levels of CD68 positive area, TNF-α and IL-1β in 24 h after reperfusion and 7 days after reperfusion in normal cerebral ischemia group, diabetic cerebral ischemia group and diabetic cerebral ischemia + 15 d-PGJ2 group were significantly higher than those in sham-operated group (P <0.05). The levels of CD68 positive area, TNF-α and IL-1β in 24 h reperfusion and 7 d reperfusion in diabetic cerebral ischemia group were significantly higher than those in control group (P <0.05). The levels of CD68 positive cells, TNF-α and IL-1β, neuron apoptosis in cerebral ischemia + 15d-PGJ2 intervention group at 24 h and 7 d after reperfusion Rate was lower than the non-intervention group (P <0.05). Conclusion Compared with the normal group, the positive rate of CD68 in diabetic cerebral ischemia group is higher, the level of TNF-α and IL-1β is higher and the apoptosis rate of nerve cells is higher. 15d-PGJ2 can reduce diabetic cerebral ischemia Mouse microglia activation, reduce the secretion of inflammatory cytokines, reduce the rate of nerve cell apoptosis.