摘要
目的观察PRMT1-ADMA-DDAH1代谢轴在糖尿病大鼠胰腺中的表达,及普罗布考对其表达和胰岛功能的影响。方法将30只雄性SD大鼠分为正常对照组(C组,n=10)、糖尿病组(D组,n=10)、糖尿病普罗布考干预组(P组,n=10)。用高脂高糖喂养联合腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型,干预组以普罗布考500mg/(kg.d)灌胃。药物干预8周后收集3组大鼠胰腺组织,ELISA法检测ADMA的浓度,用RT-PCR检测PRMT1、DDAH1 mRNA的表达,Western blot观察PRMT1蛋白的表达,免疫组化法观察胰岛素和PRMT1的表达;同时采集3组大鼠的血液标本,测定胰岛功能指标空腹血糖(FBG)、空腹血胰岛素(FINS)及氧化应激指标丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的水平。结果 C、D、P组大鼠胰腺组织中ADMA的浓度分别为(464.50±53.83)、(1255.34±188.00)、(891.72±52.10)μmol/L,D组、P组明显高于C组(P<0.01),P组显著低于D组(P<0.01);D组和P组大鼠的PRMT1 mRNA表达明显高于C组(P<0.01)、但P组明显低于D组(P<0.01),与C组相比,D、P组大鼠DDAH1的mRNA表达明显降低(P<0.01),与D组相比,P组中DDAH1的mRNA表达明显升高(P<0.01);Western blot和免疫组化的结果显示,D组与P组的PRMT1蛋白表达明显高于C组(P<0.01);而P组又明显低于D组(P<0.01)。结论糖尿病状态下,大鼠胰腺组织的PRMT1-ADMA-DDAH1代谢轴中PRMT1的表达升高,DDAH1的表达降低,导致ADMA的浓度升高,加剧氧化应激,使胰岛功能下降。而普罗布考通过其抗氧化作用,可以在一定程度上调节糖尿病状态下胰腺的PRMT1-ADMA-DDAH1代谢轴的表达,从而抑制氧化应激,改善胰岛功能。
Objective To study the expression of pancreatic PRMT1-ADMA-DDAH1 axis in rats with diabetics, and the effect of probucol on its expression and function of pancreatic islets. Methods Thirty rats were divided into control group, diabetes group, and probucol group, 10 rats in each group. A diabetic rat model was established by intra-peritoneal injection with streptozotocin （STZ） and breeding with high fat and glucose. Rats in probucol group were lavaged with probucol at 500 mg/（kg·d）. Pancreatic tissues were collected from the 3 groups 8 weeks after drug intervention. ADMA level in pancreas was measured by ELISA. Expression of PRMT1 and DDAH1 mRNA was detected by RT-PCR. Expression of PRMT1 protein in pancreas was detected by Western blotting. Expression of insulin and PRMT1 in pancreas was detected by immunohistochemistry （IHC）. Levels of FBG, FINS, MDA, SOD, and CAT in blood samples were measured. Results The ADMA level was 464.50±53.83, 1 255.34±188.00, and 891.72±52.10 μmol/L, respectively, in the 3 groups, which was higher in diabetes and probucol groups than in control group, and lower in probucol group than in diabetes group （P0.01）. The expression level of PRMT1 mRNA was significantly higher in diabetes and probucol groups than in control group （P0.01）. Western blotting and IHC showed that the expression level of DDAH1 in pancreatic PRMT1-ADMA-DDAH1 axis was higher in diabetes group than in control group and significantly lower in probucol group than in diabetes group （P0.01）. Conclusion PRMT1 is highly expressed while DDAH1 is lowly expressed in pancreatic PRMT1-ADMA-DDAH1 axis of patients with diabetes, leading to increased ADMA concentrations, oxidative stress and decreased function of pancreatic islets. Probucol can regulate the expression of pancreatic PRMT1-ADMA-DDAH1 axis, thus inhibiting oxidative stress and improving the function of pancreatic islets.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第23期2497-2502,共6页
Journal of Third Military Medical University
