CAJ | 학술논문

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(0.1 mmol/L,pH=4.5),按60mg/kg腹腔内注射,制备糖尿病模型,空腹血糖维持在13.9 mmol/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(n=17,未给予其他干预措施,正常饲养)和吡咯烷二硫基甲酸酯(核因子κB活性抑制剂)干预组[n=22,腹腔内注射吡咯烷二硫基甲酸酯(剂量20mg/kg),2次/d].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. 揹景:目前已知血管緊張素Ⅱ在糖尿病腎病髮病中起重要作用.因此,覈因子κB對糖尿病大鼠腎組織血管緊張素繫統可能有調節作用.目的:觀察抑製覈因子κB活性與糖尿病大鼠腎組織血管緊張素Ⅱ及其1型受體mRNA錶達的關繫. 設計:完全隨機分組設計,對照實驗.材料:實驗于2000-03/04在中山醫科大學實驗動物中心完成.選用51隻純種清潔級雄性Wistar大鼠.方法:①對其中39隻大鼠進行造模,採用鏈脲佐菌素溶于枸櫞痠緩遲液(0.1 mmol/L,pH=4.5),按60mg/kg腹腔內註射,製備糖尿病模型,空腹血糖維持在13.9 mmol/L以上則模型製備成功.隨機將造模後39隻大鼠分為3組:模型組(n=17,未給予其他榦預措施,正常飼養)和吡咯烷二硫基甲痠酯(覈因子κB活性抑製劑)榦預組[n=22,腹腔內註射吡咯烷二硫基甲痠酯(劑量20mg/kg),2次/d].其餘12隻為正常對照組,未造成糖尿病模型,正常飼養.②各組飼養18週後取齣腎髒,電泳遷移率變動分析技術檢測覈因子κB活性,採用反轉錄聚閤酶鏈反應法檢測血管緊張素Ⅱ1型受體mRNA錶達,採用放射免疫分析法檢測血管緊張素Ⅰ與血管緊張素Ⅱ含量.37℃水浴後的血管緊張素Ⅰ水平減去4℃檢測的血管緊張素Ⅰ水平則為腎素活性.③計量資料差異性比較採用單因素方差分析,非正態分佈資料經正態轉換後再作統計學處理.主要觀察指標:各組大鼠腎組織血管緊張素Ⅰ,Ⅱ含量和腎素、覈因子κB活性及血管緊張素Ⅱ1型受體mRNA錶達比較.結果:正常對照組、模型組、吡咯烷二硫基甲痠酯榦預組大鼠各脫失1,6,13隻,進入結果分析11,11,9隻.①覈因子κB活性:模型組明顯高于正常對照組和吡咯烷二硫基甲痠酯榦預組(P＜0.01),正常對照組與吡咯烷二硫基甲痠酯榦預組相近.②腎組織腎素活性:3組相近.③腎組織血管緊張素Ⅰ含量:模型組明顯高于正常對照組和吡咯烷二硫基甲痠酯組(P＜0.01).④腎組織血管緊張素Ⅱ含量:模型組與正常對照組相近,吡咯烷二硫基甲痠酯組明顯低于模型組和正常對照組(P＜0.01).血管緊張素Ⅱ1型受體mRNA錶達:模型組明顯低于正常對照組(P＜0.01);吡咯烷二硫基甲痠酯組明顯低于模型組和正常對照組(P＜0.01).結論:糖尿病大鼠腎組織覈因子κB活性增加,抑製覈因子κB活性後可導緻糖尿病大鼠腎組織血管緊張素Ⅱ水平及血管緊張素Ⅱ1型受體mRNA錶達下降.
배경:목전이지혈관긴장소Ⅱ재당뇨병신병발병중기중요작용.인차,핵인자κB대당뇨병대서신조직혈관긴장소계통가능유조절작용.목적:관찰억제핵인자κB활성여당뇨병대서신조직혈관긴장소Ⅱ급기1형수체mRNA표체적관계. 설계:완전수궤분조설계,대조실험.재료:실험우2000-03/04재중산의과대학실험동물중심완성.선용51지순충청길급웅성Wistar대서.방법:①대기중39지대서진행조모,채용련뇨좌균소용우구연산완충액(0.1 mmol/L,pH=4.5),안60mg/kg복강내주사,제비당뇨병모형,공복혈당유지재13.9 mmol/L이상칙모형제비성공.수궤장조모후39지대서분위3조:모형조(n=17,미급여기타간예조시,정상사양)화필각완이류기갑산지(핵인자κB활성억제제)간예조[n=22,복강내주사필각완이류기갑산지(제량20mg/kg),2차/d].기여12지위정상대조조,미조성당뇨병모형,정상사양.②각조사양18주후취출신장,전영천이솔변동분석기술검측핵인자κB활성,채용반전록취합매련반응법검측혈관긴장소Ⅱ1형수체mRNA표체,채용방사면역분석법검측혈관긴장소Ⅰ여혈관긴장소Ⅱ함량.37℃수욕후적혈관긴장소Ⅰ수평감거4℃검 측적혈관긴장소Ⅰ수평칙위신소활성.③계량자료차이성비교채용단인소방차분석,비정태분포자료경정태전환후재작통계학처리.주요관찰지표:각조대서신조직혈관긴장소Ⅰ,Ⅱ함량화신소、핵인자κB활성급혈관긴장소Ⅱ1형수체mRNA표체비교.결과:정상대조조、모형조、필각완이류기갑산지간예조대서각탈실1,6,13지,진입결과분석11,11,9지.①핵인자κB활성:모형조명현고우정상대조조화필각완이류기갑산지간예조(P＜0.01),정상대조조여필각완이류기갑산지간예조상근.②신조직신소활성:3조상근.③신조직혈관긴장소Ⅰ함량:모형조명현고우정상대조조화필각완이류기갑산지조(P＜0.01).④신조직혈관긴장소Ⅱ함량:모형조여정상대조조상근,필각완이류기갑산지조명현저우모형조화정상대조조(P＜0.01).혈관긴장소Ⅱ1형수체mRNA표체:모형조명현저우정상대조조(P＜0.01);필각완이류기갑산지조명현저우모형조화정상대조조(P＜0.01).결론:당뇨병대서신조직핵인자κB활성증가,억제핵인자κB활성후가도치당뇨병대서신조직혈관긴장소Ⅱ수평급혈관긴장소Ⅱ1형수체mRNA표체하강.
BACKGROUND: Nowadays, angiotensin Ⅱ plays an important role in onset of diabetic nephropathy. Therefore, the nuclear factor-κB may have adjustive effects on angiotonin system of kidney tissue of diabetic rats. OBJECTIVE: To observe the relationship of activity of inhibitive nuclear factor-κB with angiotensin Ⅱ and its type 1 receptor mRNA expression of renal tissue of diabetic rats. DESIGN: Completely randomized group design, control experiment. MATERIALS: The experiment was conducted at the Experimental Animal Center, Sun Yat-sen University of Medical Sciences between March and April 2000. Fifty-one pure breed clean grade male Wistar rats were select ed. METHODS: ①Models were established in 39 rats. Streptozotocin dissolv ing in citric acid buffer (0.1 mmol/L,pH=4.5) were given to establish dia betic models with 60 mg/kg intraperitoneal injection. If the fasting blood glucose maintained above 13.9 mmol/L, the establishment of models was successful. The thirty-nine rats were randomly assigned into 3 groups: model group (n=17, without other interventional measure, feeding normally) and pyrrolidine dithiocar2. Bamate (PDTC) (active inhibitor of nuclear fac tor-κB) interventional group [n=22, PDTC at the dose of 20 mg/kg were given with intraperitoneal injection, twice a day]. Other 12 rats were as normal control group, did not make into diabetic models with normal breeding. ②After feeding for 18 weeks kidneys were got in every group. The activity of nuclear factor-κB was detected with electrophoretic mobility shift assay. The expression of type 1 receptor mRNA of angiotensin Ⅱ was measured with reverse transcription polymerase chain reaction (RT-PCR). Contents of angiotonin Ⅰ and angiotensin Ⅱ were tested with Radio Im munoassay (RIA). Activity of rennin was referred to that the result of the level of angiotonin Ⅰ at 37 ℃ water bath subduced to that at 4 ℃. ③Dif ference of measurement data was compared with single factor analysis of variance. After normal transformation, the non-normal distribution data were conducted with statistical disposal. MAIN OUTCOME MEASURES: Comparison of contents of angiotensin Ⅰ and Ⅱ, activities of rennin and nuclear factor-κB and expression of type 1 receptor mRNA of angiotensin Ⅱ in renal tissues of rats of each group. RESULTS: In the normal control group, model group and PDTC interven tional group 1, 6 and 13 rats were dropped out, respectively, so 11, 11 and 9 rats in each group were involved in the result analysis. ①Activity of nu clear factor-κB: It was higher significantly in the model group than that in the normal control group and PDTC interventional group (P ＜ 0.01 ). It was similar between the normal control group and the PDTC interventional group. ②Activity of rennin of renal tissue: It was similar among the 3 groups. ③Content of angiotonin Ⅰ of renal tissue: It was higher obviously in the model group that that in the normal control group and the PDTC interventional group (P ＜ 0.01 ). ④Content of angiotensin Ⅱ in renal tissue: It was similar between the model group and the normal control group. It was lower markedly in the PDTC interventional group than that in the model group and the normal control group (P ＜ 0.01 ). Expression of type 1 receptor mRNA of angiotensin Ⅱ: It was lower remarkably in the model group than that in the normal control group (P ＜ 0.01 ). It was lower dis tinctly in the PDTC interventional group than that in the model group and the normal control group (P ＜ 0.01 ). CONCLUSION: The increase of activity of nuclear factor-κB in renal tissue of diabetic rats can inhibit the activity of nuclear factor-κB, which will induce the reduction of the level of angiotensin Ⅱ and expression of type 1 receptor mRNA of angiotensin Ⅱ in renal tissue of diabetic rats.