2型糖尿病大鼠脑血管内皮细胞CD34 TNF-αVEGF水平变化及氨基胍干预的保护作用研究

Type 2 diabetic rat brain vascular endothelial cells CD34 expression and TNF-α,VEGF levels changes and the protective effect of aminoguanidine intervention

目的研究2型糖尿病大鼠海马区脑血管内皮细胞CD34表达及血管内皮细胞炎症因子肿瘤坏死因子-α、血管内皮生长因子水平变化及氨基胍干预的脑保护作用及机制.方法将24只SD大鼠按照随机数字表法分为对照组、模型组、治疗组,每组8只,于分组后12周、16周两个时间段观察;对照组给予普通饮食喂养,其余2组按照文献建立糖尿病大鼠模型,模型组及治疗组给予高脂高糖饮食喂养6周,用ONETOUCH血糖仪测定血糖,以随机非空腹血糖>16.7mmol·L-1定为糖尿病模型,造模成功后,治疗组给予氨基胍溶液每天灌胃,对照组及模型组给予等量生理盐水灌胃处理.采用免疫组化方法测定海马区脑血管密度CD34表达变化,采取酶联免疫吸附法测定海马区血管内皮细胞肿瘤坏死因子-α、血管内皮生长因子水平变化.结果与对照组相比,模型组和治疗组脑血管内皮细胞CD34表达均显著增加(P<0.01),肿瘤坏死因子-α、血管内皮生长因子水平均显著升高(P<0.01);模型组16周CD34表达较12周显著增加(P<0.01),肿瘤坏死因子-α、血管内皮生长因子水平较12周显著升高(P<0.05或0.01);治疗组各时间点CD34表达均显著少于模型组(P<0.01),肿瘤坏死因子-α、血管内皮生长因子水平均显著低于模型组(P<0.01).结论CD34表达及血管内皮细胞炎症因子肿瘤坏死因子-α、血管内皮生长因子水平变化均参与及影响2型糖尿病大鼠脑血管重塑过程,氨基胍干预对2型糖尿病大鼠脑保护作用是通过抑制肿瘤坏死因子-α、血管内皮生长因子的产生等途径改善糖尿病脑血管重塑,发挥脑保护作用.

Objective To observe the type 2 diabetic rat brain vascular endothelial cells CD34 expression and TNF-a,VEGF levels changes and the protective effect of aminoguanidine intervention.Methods 24 cases of SD rats were randomly divided into control group,model group(diabetes group)and treatment group(aminoguanidine intervention group),8 cases each group.After grouping,observe in two time peri-ods of 12 weeks and 16 weeks.The control group was given a normal diet and the remaining 2 groups es-tablished a diabetic rat model according to the literature.The model group and the intervention group were fed a high-fat and high-sugar diet for 6 weeks,and the blood glucose was measured with the ONETOUCH blood glucose meter.Randomized non-fasting blood glucose>16.7 mmol.L.-1 was used as the diabetes model.After successful modeling,the treatment group was given intragastric administration of aminogua-nidine solution every day,and the control group and model group were given intragastric administration of normal saline.Immunohistochemical method was used to determine the expression of CD34 in the cerebral blood vessel density in hippocampus,and ELISA was used to determine the changes of tumor necro-sis factor-a and vascular endothelial growth factor in vascular endothelial cells in hippocampus.Results Compared with the control group,the expression of CD34 in cerebral vascular endothelial cells of the mod-el group and the treatment group were significantly increased(P<0.01),and the levels of TNF-a and VEGF were significantly increased(P<0.01).In the model group,the expression of CD34 increased sig-nificantly at 16 weeks compared to 12 weeks(P<0.01),the levels of TNF-a and VEGF were significantly higher than those at 12 weeks(P<0.05 or 0.01).The expression of CD34 at each time point in the treat-ment group was significantly less than that in the model group(P<0.01),and the levels of TNF-a and VEGF were significantly lower than those in the model group(P<0.01).Conclusions The expression of CD34 and the changes of vascular endothelial cell inflammatory factors,TNF-a and VEGF levels are in-volved in and affect the process of cerebral vascular remodeling in type 2 diabetic rats.The protective effect of aminoguanidine on the brain of type 2 diabetic rats is to improve the remodeling of diabetic cere-bral blood vessels by inhibiting the production of TNF-a and VEGF and play a role in brain protection.