普罗布考对2型糖尿病大鼠胰岛氧化应激及功能的影响
摘要
目的:建立2型糖尿病动物模型,观察糖尿病大鼠胰腺硝基酪氨酸和8-OH脱氧鸟苷等氧化应激指标表达水平的变化,并通过普罗布考的早期干预,来观察普罗布考对糖尿病氧化应激及胰腺功能的影响。
方法:雄性8周龄Wister大鼠,随机分为C组(n=7),D组(n=9)和P组(n=7)。其中,C组为正常对照组,普通饲料喂养4周后腹腔内注射柠檬酸缓冲液;D组为2型糖尿病模型组,高脂高糖饲料喂养4周后,按30mg/Kg体重剂量腹腔内注射STZ溶液;P组为普罗布考干预的2型糖尿病模型组,在喂养高脂高糖饲料的同时灌胃以普罗布考,4周后按30mg/Kg体重剂量腹腔内注射STZ溶液。10周后处死,测定空腹血糖(FPG)、空腹胰岛素(FINS)、总胆固醇(TC)、甘油三脂血脂(TG)、MDA及SOD,计算胰岛素敏感指数(ISI),用免疫组化的方法观察NT、8-OHdG及胰岛素的表达。
结果:D组及P组大鼠的FPG、TG、TC及MDA水平较C组均明显增加(P<0.01),NT和8-OHdG表达的平均光密度也明显高于C组(P<0.01),但P组上述指标的测定又显著低于D组(P<0.01);D组和P组大鼠的SOD水平、ISI及胰岛素阳性面积百分比均明显低于C组(P<0.01),P组这些指标的测定显著高于D组(P<0.01);C组和P组的FINS比较没有显著统计学差异,但都明显高于D组(P<0.01)。
结论:糖尿病状态下,氧化应激明显增加,胰腺功能明显减退,早期给与普罗布考干预,在降血脂和抗氧化作用的同时一定程度上降低了血糖,保护了胰腺功能,说明氧化应激可能在2型糖尿病的发病机制中起重要作用。
Objective: To establish the animal model of type 2 diabetes mellitus and to observe the levels of nitrotyrosine and 8-hydroxy-2'-deoxyguanosine, which were the biomarkers of oxidative stress. And to observe the effects on oxidative stress and pancreatic islets function in diabetic rats by treated with probucol in early stage.
Methods: The male Wister rats of 8 weeks old randomly devided into C (n=7),D (n=9) and P (n=7). C was the normal control group, feeding with regular diets ,4 weeks later injected citrate buffer liquid in abdominal cavity; D was the type 2 dibetes model group, feeding with the diets enriched with high-fat and high-sucrose, 4 weeks later injected the aqua of streptozotocin(STZ) at a dose of 30mg/Kg body weight in abdominal cavity; P was the probucol group, feeding with the diets enriched with high-lipid、high-sucrose and probucol, 4 weeks later injected the aqua of streptozotocin(STZ) at a dose of 30mg/Kg body weight. 10 weeks later, measured the fasting plasma glucose(FPG), fasting insuline(FINS), total cholesterol(TC), triglyceride(TG),MDA and SOD,and calculated the insulin sensitivity index(ISI). To observe the expression of NT、8-OHdG and insulin by immunohistochemistric method.
Results: The FPG、TG、TC and MDA level of C and D group were significantly increased(P<0.01), and the average optical density of NT and 8-OHdG were much higher than C group(P<0.01), but the markers of P group decreased significantly to compared with D group(P<0.01). The level of SOD, value of ISI and percentage of the insulin positive area in the pancreatic islets of D and P group were much lower than C group(P<0.01), however, the markers of P group were higher than D group apparently(P<0.01). The FINS level of C and P group had no significant difference, whereas they increased significantly to compared with D group. Conclusions: The activities of oxidative stress were enhanced and function of pancreatic islets degenerated apparently in diabetes condition. With the treatment of probucol in early stage, the levels of TG and TC decreased significantly, meanwhile partially reduced the FPG and restored the pancreatic islets function acting through as an antioxidant. This suggests that oxidative stress may play an important role in the mechanism of type 2 diabetes mellitus.
方法:雄性8周龄Wister大鼠,随机分为C组(n=7),D组(n=9)和P组(n=7)。其中,C组为正常对照组,普通饲料喂养4周后腹腔内注射柠檬酸缓冲液;D组为2型糖尿病模型组,高脂高糖饲料喂养4周后,按30mg/Kg体重剂量腹腔内注射STZ溶液;P组为普罗布考干预的2型糖尿病模型组,在喂养高脂高糖饲料的同时灌胃以普罗布考,4周后按30mg/Kg体重剂量腹腔内注射STZ溶液。10周后处死,测定空腹血糖(FPG)、空腹胰岛素(FINS)、总胆固醇(TC)、甘油三脂血脂(TG)、MDA及SOD,计算胰岛素敏感指数(ISI),用免疫组化的方法观察NT、8-OHdG及胰岛素的表达。
结果:D组及P组大鼠的FPG、TG、TC及MDA水平较C组均明显增加(P<0.01),NT和8-OHdG表达的平均光密度也明显高于C组(P<0.01),但P组上述指标的测定又显著低于D组(P<0.01);D组和P组大鼠的SOD水平、ISI及胰岛素阳性面积百分比均明显低于C组(P<0.01),P组这些指标的测定显著高于D组(P<0.01);C组和P组的FINS比较没有显著统计学差异,但都明显高于D组(P<0.01)。
结论:糖尿病状态下,氧化应激明显增加,胰腺功能明显减退,早期给与普罗布考干预,在降血脂和抗氧化作用的同时一定程度上降低了血糖,保护了胰腺功能,说明氧化应激可能在2型糖尿病的发病机制中起重要作用。
Objective: To establish the animal model of type 2 diabetes mellitus and to observe the levels of nitrotyrosine and 8-hydroxy-2'-deoxyguanosine, which were the biomarkers of oxidative stress. And to observe the effects on oxidative stress and pancreatic islets function in diabetic rats by treated with probucol in early stage.
Methods: The male Wister rats of 8 weeks old randomly devided into C (n=7),D (n=9) and P (n=7). C was the normal control group, feeding with regular diets ,4 weeks later injected citrate buffer liquid in abdominal cavity; D was the type 2 dibetes model group, feeding with the diets enriched with high-fat and high-sucrose, 4 weeks later injected the aqua of streptozotocin(STZ) at a dose of 30mg/Kg body weight in abdominal cavity; P was the probucol group, feeding with the diets enriched with high-lipid、high-sucrose and probucol, 4 weeks later injected the aqua of streptozotocin(STZ) at a dose of 30mg/Kg body weight. 10 weeks later, measured the fasting plasma glucose(FPG), fasting insuline(FINS), total cholesterol(TC), triglyceride(TG),MDA and SOD,and calculated the insulin sensitivity index(ISI). To observe the expression of NT、8-OHdG and insulin by immunohistochemistric method.
Results: The FPG、TG、TC and MDA level of C and D group were significantly increased(P<0.01), and the average optical density of NT and 8-OHdG were much higher than C group(P<0.01), but the markers of P group decreased significantly to compared with D group(P<0.01). The level of SOD, value of ISI and percentage of the insulin positive area in the pancreatic islets of D and P group were much lower than C group(P<0.01), however, the markers of P group were higher than D group apparently(P<0.01). The FINS level of C and P group had no significant difference, whereas they increased significantly to compared with D group. Conclusions: The activities of oxidative stress were enhanced and function of pancreatic islets degenerated apparently in diabetes condition. With the treatment of probucol in early stage, the levels of TG and TC decreased significantly, meanwhile partially reduced the FPG and restored the pancreatic islets function acting through as an antioxidant. This suggests that oxidative stress may play an important role in the mechanism of type 2 diabetes mellitus.
引文
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