摘要
目的:观察抑制核因子κB的活性对糖尿病大鼠肾组织诱导型一氧化氮合酶表达的影响及与肾功能损害的关系。方法:实验于2005-09/2006-07在辽宁医学院生物化学实验室完成。将45只SD大鼠随机分为正常对照组、模型组和PDTC干预组3组,每组15只。模型组和PDTC干预组大鼠腹腔注射链脲佐菌素60mg/kg制备大鼠糖尿病模型,正常对照组注射相应体积的柠檬酸缓冲液。造模成功后PDTC干预组腹腔注射核因子κB活性抑制剂吡咯烷二硫基甲酸酯(PDTC),2次/d,剂量按每周20mg/kg。于注射链脲佐菌素12周末检测血清肌酐、尿素氮和24h尿蛋白定量,然后处死大鼠,留取肾标本,观测肾脏的病理改变并以Western-blot法检测核因子κB的含量,免疫组化法分析核因子κBP65及诱导型一氧化氮合酶的表达,同时应用直线相关法分析这2项指标的表达与生化指标间的关系。结果:30只大鼠进入结果分析。①模型组肾组织中核因子κBP65阳性小管数多于正常对照组和PDTC干预组[(49.35±3.87)%,(1.76±0.82)%,(24.61±3.05)%,P<0.01],PDTC干预组高于正常对照组(P<0.01)。②模型组肾组织中诱导型一氧化氮合酶阳性小管数多于正常对照组和PDTC干预组[(51.63±4.75)%,(2.52±0.93)%,(26.41±2.86)%,P<0.01],PDTC干预组高于正常对照组(P<0.05)。③模型组核因子κB的灰度值明显高于正常对照组和PDTC干预组(0.2995±0.006,0.1375±0.005,0.2045±0.004,P<0.01),PDTC干预组高于正常对照组(P<0.05)。④模型组和PDTC干预组血清肌酐、尿素氮和24h尿蛋白均高于正常对照组(P<0.01,0.05),但PDTC干预组几项指标均低于模型组(P<0.01)。⑤直线相关分析结果显示核因子κB的表达与诱导型一氧化氮合酶的表达、肾功能损害呈显著相关。结论:抑制核因子κB活性能使诱导型一氧化氮合酶的表达减少进而降低肾功能损害,提示大鼠糖尿病肾病的发生可能与肾组织中核因子κB活化介导诱导型一氧化氮合酶表达上调有关。
AIM. To observe the effect of the inhibition of nuclear factor-kappa B （NF-κB） on the expression of inducible nitricoxide synthase （iNOS） in the renal tissue of rats with diabetes and its correlation with renal function injury.
METHODS： The experiment was performed in the biochemical laboratory of Liaoning Medical College from September 2005 to July 2006. Forty-five Sprague-Dawiey rats were randomly divided into 3 groups： control group, model group and pyrrolidinedithiocerbamate （PDTC） group with 15 rats in each group. The rats of the latter two groups were intraperitoneally injected 60 mg/kg streptozotocin to prepare the rat models of diabetes; the control group was given matching citrate buffer solution. After successfully modeling, PDTC group was injected PDTC, an inhibitor of NF-κB activity, twice daily at a dose of 20 mg/kg per week. At the end of the 12^th week of streptozotocin injection, the serum creatinine, urea nitrogen and quantities of 24-hour urine albumin were detected, and the kidneys were taken after the rats were killed to observe the pathological changes. The content of NF-κB was detected by Western-blot; the expression of NF-κB P65 and iNOS was detected by immunohistochemistry, and the correlation between the two indexes and biochemical indexes was analyzed with linear correlation method.
RESULTS： Thirty rats were involved in the result analysis. （1）The number of NF-κB P65 positive tubulus in renal tissue of the model group was more than the control group and PDTC group [（49.35±3.87）%, （1.76±0.82）%, （24.61±3.05）%, P 〈 0.01], and PDTC group was higher than the control group （P〈 0.01）. （2）The number of iNOS positive tubulus in renal tissue of the model group was more than the control group and PDTC group [（51.63±4.75）%, （2.52±0.93）%, （26.41± 2.86）%, P 〈 0.01], and the PDTC group was higher than the control group （P 〈 0.05）. （3）The gray degree values of the model group was obviously higher than the control group and PDTC group （0.299 5±0.006, 0.137 5±0：005, 0.204 5± 0.004, P〈 0.01）, and the PDTC group was higher than the control group （P〈 0.05）. （4）The serum creatinine, urea nitrogen and quantities of 24-hour urine albumin of the model and PDTC groups were higher than the control group （P 〈 0.01, 0.05）, but all the indexes of the PDTC group were lower than the model group （P 〈 0.01）. （5）Linear correlation analysis showed that the expression of NF-κB had a strong correlation with the expression of iNOS and renal functional lesion, respectively.
CONCLUSION： The inhibition of NF-κB activity could decrease the expression of iNOS and reduce the renal functional damage, indicating that the occurrence of diabetic nephrepathy in rats may be related with the intervention of NF-κB activity on the upregulation of the expression of iNOS.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第6期1071-1074,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
