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背景:目前已知血管紧张素Ⅱ在糖尿病肾病发病中起重要作用。因此,核因子κB对糖尿病大鼠肾组织血管紧张素系统可能有调节作用。目的:观察抑制核因子κB活性与糖尿病大鼠肾组织血管紧张素Ⅱ及其1型受体mRNA表达的关系。设计:完全随机分组设计,对照实验。材料:实验于2000-03/04在中山医科大学实验动物中心完成。选用51只纯种清洁级雄性Wistar大鼠。方法:①对其中39只大鼠进行造模,采用链脲佐菌素溶于枸橼酸缓冲液穴0.1mmol/L,pH=4.5雪,按60mg/kg腹腔内注射,制备糖尿病模型,空腹血糖维持在13.9mmol/L以上则模型制备成功。随机将造模后39只大鼠分为3组:模型组穴n=17,未给予其他干预措施,正常饲养雪和吡咯烷二硫基甲酸酯(核因子κB活性抑制剂)干预组眼n=22,腹腔内注射吡咯烷二硫基甲酸酯(剂量20mg/kg),2次/d演。其余12只为正常对照组,未造成糖尿病模型,正常饲养。②各组饲养18周后取出肾脏,电泳迁移率变动分析技术检测核因子κB活性,采用反转录聚合酶链反应法检测血管紧张素Ⅱ1型受体mRNA表达,采用放射免疫分析法检测血管紧张素Ⅰ与血管紧张素Ⅱ含量。37℃水浴后的血管紧张素Ⅰ水平减去4℃检测的血管紧张素Ⅰ水平则为肾素活性。③计量资料差异性比较采用单因素方差分析,非正态分布资料经正态转换后再作统计学处理。主要观察指标:各组大鼠肾组织血管紧张素Ⅰ,Ⅱ含量和肾素、核因子κB活性及血管紧张素Ⅱ1型受体mRNA表达比较。结果:正常对照组、模型组、吡咯烷二硫基甲酸酯干预组大鼠各脱失1,6,13只,进入结果分析11,11,9只。①核因子κB活性:模型组明显高于正常对照组和吡咯烷二硫基甲酸酯干预组(P<0.01),正常对照组与吡咯烷二硫基甲酸酯干预组相近。②肾组织肾素活性:3组相近。③肾组织血管紧张素Ⅰ含量:模型组明显高于正常对照组和吡咯烷二硫基甲酸酯组(P<0.01)。④肾组织血管紧张素Ⅱ含量:模型组与正常对照组相近,吡咯烷二硫基甲酸酯组明显低于模型组和正常对照组(P<0.01)。血管紧张素Ⅱ1型受体mRNA表达:模型组明显低于正常对照组(P<0.01);吡咯烷二硫基甲酸酯组明显低于模型组和正常对照组(P<0.01)。结论:糖尿病大鼠肾组织核因子κB活性增加,抑制核因子κB活性后可导致糖尿病大鼠肾组织血管紧张素Ⅱ水平及血管紧张素Ⅱ1型受体mRNA表达下降。
BACKGROUND: Angiotensin Ⅱ is currently known to play an important role in the pathogenesis of diabetic nephropathy. Therefore, NF-kappaB may regulate the renal tissue angiotensin system in diabetic rats. OBJECTIVE: To observe the relationship between the inhibition of nuclear factor kappa B (NF - κB) activity and the expression of angiotensin Ⅱ and its type 1 receptor mRNA in the kidney of diabetic rats. Design: Complete randomized block design, control experiment. Materials: The experiment was performed at Experimental Animal Center of Sun Yat-sen University from March to April in 2000. Fifty purebred male Wistar rats were selected. Methods: ①Among 39 rats, streptozotocin was dissolved in citrate buffer 0.1mmol / L, pH = 4.5, and then injected into the abdominal cavity of 60mg / kg to prepare diabetic model. Fasting Blood glucose remained above 13.9mmol / L model was successfully prepared. A total of 39 rats were randomly divided into 3 groups: model group, n = 17, no other intervention was given, and snow and pyrrolidine dithiocarbamate (inhibitor of nuclear factor κB) n = 22, intraperitoneal injection of pyrrolidine dithiocarbamate (dose 20mg / kg), 2 times / d played. The remaining 12 as a normal control group, did not cause diabetes model, normal breeding. ② After 18 weeks of feeding, the kidneys were removed and the activity of NF-κB was detected by electrophoretic mobility shift assay. The expression of angiotensin Ⅱ type 1 receptor mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) Angiotensin Ⅰ and angiotensin Ⅱ content. The level of angiotensin I after 37 ° C water bath minus the level of angiotensin I detected at 4 ° C was renin activity. (3) The differences of measurement data were compared using one-way analysis of variance (ANOVA). The data of non-normal distribution were normalized and then processed statistically. MAIN OUTCOME MEASURES: The levels of angiotensin Ⅰ, Ⅱ and the expressions of renin and nuclear factor κB and the mRNA expression of angiotensin Ⅱ type 1 receptor in rat kidney were compared. Results: The normal control group, model group, pyrrolidine dithioate intervention group lost 1,6,13 rats, 11,1,9 into the results of analysis. ① The activity of NF-κB: The model group was significantly higher than the normal control group and pyrrolidine dithiocarbamate intervention group (P <0.01). The normal control group was similar to the pyrrolidine dithiocarbamate intervention group. Renal renin activity: similar to 3 groups. ③ The content of angiotensin Ⅰ in kidney tissue: The model group was significantly higher than the normal control group and pyrrolidine dithiocarbamate group (P <0.01). ④ The content of angiotensin Ⅱ in renal tissue: Compared with the normal control group, the pyrrolidine dithiocarbamate group in the model group was significantly lower than the model group and the normal control group (P <0.01). The mRNA expression of angiotensin Ⅱ type 1 receptor in model group was significantly lower than that in normal control group (P <0.01). Pyrrolidine dithiocarbamate group was significantly lower than model group and normal control group (P <0.01). CONCLUSION: The increase of nuclear factor kappa B in renal tissue and the decrease of the expression of angiotensin Ⅱ type Ⅱ receptor and angiotensin Ⅱ type 1 receptor in renal tissue of diabetic rats can be induced by inhibiting the activity of nuclear factor.