中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2008年
15期
2988-2992
,共5页
陈泽君%王平%黄颂敏%曹舸%郭华%程弓%佘宁兰
陳澤君%王平%黃頌敏%曹舸%郭華%程弓%佘寧蘭
진택군%왕평%황송민%조가%곽화%정궁%사저란
糖尿病肾病%血管内皮生长因子类%受体,血管紧张素,2型/拮抗剂和抑制剂
糖尿病腎病%血管內皮生長因子類%受體,血管緊張素,2型/拮抗劑和抑製劑
당뇨병신병%혈관내피생장인자류%수체,혈관긴장소,2형/길항제화억제제
背景:已有实验提示,血管内皮生长因子及其受体系统可能参与糖尿病肾病的发生发展.目的:实验进一步验证血管内皮生长因子及其受体Flk-1在糖尿病鼠肾脏中的表达变化及厄贝沙坦对其影响和可能的机制.设计:随机对照动物实验.单位:四川大学华西医院.材料:雄性封闭群SD大鼠18只,体质量150-200 g,标准化饲养.方法:实验于2006-08/2007-04在四川大学华西医院实验室完成.建立糖尿病肾病大鼠模型,分为糖尿病肾病组、厄贝沙坦干预组和正常对照组,每组6只大鼠.采用10 g/L链脲霉素按55 mg/kg体质量一次性腹腔内注射诱导造模,对照组给予相同剂量的枸橼酸缓冲液,干预组在成模后按3 mg/(kg·d)给予厄贝沙坦.采用反转录聚合酶链式反应技术以及免疫组织化学技术检测血管内皮生长因子和Flk-1的表达,检测尿蛋白、肾小球面积和体积,并分析数据相关性.主要观察指标:①肾脏病理以及尿蛋白、肾小球面积和体积.②肾脏血管内皮生长因子与Flk-l mRNA表达.③肾脏免疫组织化学检查.④相关分析.结果:①肾脏苏木精-伊红染色病理榆查显示糖尿病肾病组出现明显.肾小球增大,系膜基质增多,系膜细胞增多,小管扩张,干预组病变轻于糖尿病肾病组.精尿病肾病组尿蛋白显著高于对照组(P＜0.01);糖尿病肾病组肾重/体质量、肾小球面积和体积明显高于对照组(P＜0.01),干预组蛋白尿、肾重/体质量、肾小球面积和体积均低于糖尿病肾病组(P＜0.01).②糖尿病肾病组16周时肾脏血管内皮生长因子与Flk-1 mRNA表达均明显上调,显著高于对照组(P＜0.05);干预组16周时肾脏血管内皮生长因子与Flk-1 mRNA表达也有上调,高于对照组(P＜0.05),但低于糖尿病肾病组(P＜0.05).③糖尿病肾病组血管内皮生长因子着染明显强于对照组(P＜0.01),干预组着染也强于对照组(P＜0.01),但与糖尿病肾病相比较,干预组着染明显减少(P＜0.01).Flk-1着染也有类似变化.④血管内皮生长因子、Flk-1与尿蛋白、肾小球面积和体积呈正相关(P＜0.05).结论:血管内皮生长因子及其受体Flk-1在糖尿病肾病发病机制中起重要作用,过度表达导致肾脏损伤,血管紧张素Ⅱ受体拮抗剂厄贝沙坦具有通过抑制血管内皮生长因子和Flk-1异常表达这一非血流动力学的肾脏保护作用.
揹景:已有實驗提示,血管內皮生長因子及其受體繫統可能參與糖尿病腎病的髮生髮展.目的:實驗進一步驗證血管內皮生長因子及其受體Flk-1在糖尿病鼠腎髒中的錶達變化及阨貝沙坦對其影響和可能的機製.設計:隨機對照動物實驗.單位:四川大學華西醫院.材料:雄性封閉群SD大鼠18隻,體質量150-200 g,標準化飼養.方法:實驗于2006-08/2007-04在四川大學華西醫院實驗室完成.建立糖尿病腎病大鼠模型,分為糖尿病腎病組、阨貝沙坦榦預組和正常對照組,每組6隻大鼠.採用10 g/L鏈脲黴素按55 mg/kg體質量一次性腹腔內註射誘導造模,對照組給予相同劑量的枸櫞痠緩遲液,榦預組在成模後按3 mg/(kg·d)給予阨貝沙坦.採用反轉錄聚閤酶鏈式反應技術以及免疫組織化學技術檢測血管內皮生長因子和Flk-1的錶達,檢測尿蛋白、腎小毬麵積和體積,併分析數據相關性.主要觀察指標:①腎髒病理以及尿蛋白、腎小毬麵積和體積.②腎髒血管內皮生長因子與Flk-l mRNA錶達.③腎髒免疫組織化學檢查.④相關分析.結果:①腎髒囌木精-伊紅染色病理榆查顯示糖尿病腎病組齣現明顯.腎小毬增大,繫膜基質增多,繫膜細胞增多,小管擴張,榦預組病變輕于糖尿病腎病組.精尿病腎病組尿蛋白顯著高于對照組(P＜0.01);糖尿病腎病組腎重/體質量、腎小毬麵積和體積明顯高于對照組(P＜0.01),榦預組蛋白尿、腎重/體質量、腎小毬麵積和體積均低于糖尿病腎病組(P＜0.01).②糖尿病腎病組16週時腎髒血管內皮生長因子與Flk-1 mRNA錶達均明顯上調,顯著高于對照組(P＜0.05);榦預組16週時腎髒血管內皮生長因子與Flk-1 mRNA錶達也有上調,高于對照組(P＜0.05),但低于糖尿病腎病組(P＜0.05).③糖尿病腎病組血管內皮生長因子著染明顯彊于對照組(P＜0.01),榦預組著染也彊于對照組(P＜0.01),但與糖尿病腎病相比較,榦預組著染明顯減少(P＜0.01).Flk-1著染也有類似變化.④血管內皮生長因子、Flk-1與尿蛋白、腎小毬麵積和體積呈正相關(P＜0.05).結論:血管內皮生長因子及其受體Flk-1在糖尿病腎病髮病機製中起重要作用,過度錶達導緻腎髒損傷,血管緊張素Ⅱ受體拮抗劑阨貝沙坦具有通過抑製血管內皮生長因子和Flk-1異常錶達這一非血流動力學的腎髒保護作用.
배경:이유실험제시,혈관내피생장인자급기수체계통가능삼여당뇨병신병적발생발전.목적:실험진일보험증혈관내피생장인자급기수체Flk-1재당뇨병서신장중적표체변화급액패사탄대기영향화가능적궤제.설계:수궤대조동물실험.단위:사천대학화서의원.재료:웅성봉폐군SD대서18지,체질량150-200 g,표준화사양.방법:실험우2006-08/2007-04재사천대학화서의원실험실완성.건립당뇨병신병대서모형,분위당뇨병신병조、액패사탄간예조화정상대조조,매조6지대서.채용10 g/L련뇨매소안55 mg/kg체질량일차성복강내주사유도조모,대조조급여상동제량적구연산완충액,간예조재성모후안3 mg/(kg·d)급여액패사탄.채용반전록취합매련식반응기술이급면역조직화학기술검측혈관내피생장인자화Flk-1적표체,검측뇨단백、신소구면적화체적,병분석수거상관성.주요관찰지표:①신장병리이급뇨단백、신소구면적화체적.②신장혈관내피생장인자여Flk-l mRNA표체.③신장면역조직화학검사.④상관분석.결과:①신장소목정-이홍염색병리유사현시당뇨병신병조출현명현.신소구증대,계막기질증다,계막세포증다,소관확장,간예조병변경우당뇨병신병조.정뇨병신병조뇨단백현저고우대조조(P＜0.01);당뇨병신병조신중/체질량、신소구면적화체적명현고우대조조(P＜0.01),간예조단백뇨、신중/체질량、신소구면적화체적균저우당뇨병신병조(P＜0.01).②당뇨병신병조16주시신장혈관내피생장인자여Flk-1 mRNA표체균명현상조,현저고우대조조(P＜0.05);간예조16주시신장혈관내피생장인자여Flk-1 mRNA표체야유상조,고우대조조(P＜0.05),단저우당뇨병신병조(P＜0.05).③당뇨병신병조혈관내피생장인자착염명현강우대조조(P＜0.01),간예조착염야강우대조조(P＜0.01),단여당뇨병신병상비교,간예조착염명현감소(P＜0.01).Flk-1착염야유유사변화.④혈관내피생장인자、Flk-1여뇨단백、신소구면적화체적정정상관(P＜0.05).결론:혈관내피생장인자급기수체Flk-1재당뇨병신병발병궤제중기중요작용,과도표체도치신장손상,혈관긴장소Ⅱ수체길항제액패사탄구유통과억제혈관내피생장인자화Flk-1이상표체저일비혈류동역학적신장보호작용.
BACKGROUND: Some studies have presented that vascular endothelial growth factor and its receptor system may take part in onset and development of diabetic nephropathy (DN).OBJECTIVE: To further verify the interventional effects of irbesartan on vascular endothelial growth factor (VEGF) and its Flk-1 receptor expressions in kidney of diabetes mellitus (DM) rat, and the possible mechanism of irbesartan.DESIGN: Randomized control animal study.SETTING: West China Hospital of Sichuan University.MATERIALS: Eighteen male closed colony SD rats weighing 150-200 g were cared in standardization.METHODS: This study was performed at Laboratory of West China Hospital of Sichuan University from August 2006 to April 2007. All rats were randomly divided into a DN group, an irbesartan group and a normal control group, with 6 rats in each group. 10 g/L streptozotocin (55 mg/kg) was intraperitoneally injected to establish models; rats in the control group were administrated with the same dosage of citric acid buffer solution; rats in the irbesartan group were administrated with 3 mg/(kg·d) irbesartan after model establishment. VEGF and FIk-1 expressions were detected using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry technique; while, urine protein level, area and volume of renal glomerulus were detected, and correlations of data were analyzed.MAIN OUTCOME MEASURES: ①Renal pathological indicators, urine protein level, area and volume of renal glomerulus; ②VEGF and Flk-1 expressions;③renal immunohistochemical examination;④ correlation analysis.RESULTS:① Renal pathological examination by HE staining indicated that renal glomerulus was remarkably enlarged in the DN group; mesangial matrix was increased; mesangial cells were also increased; renal tubule was expanded. The lesions in the irbesartan group were milder compared to the DN group. Urine protein level in the DN group was significantly higher compared to the control group (P ＜ 0.01); renal weight/body mass, area and volume of renal glomerulus were significantly higher compared to the control group (P ＜ 0.01); urine protein level, renal weight/body mass, area and volume of renal glomerulus in the irbesartan group were significantly lower compared to the DN group (P ＜ 0.01). ② By the 16th week,VEGF and Flk-1 expressions in the DN group were significantly up-regulated compared to control group (P ＜ 0.05); while,VEGF and Flk-1 expressions in the irbesartan group were also up-regulated compared to the control group by the 16th week (P ＜ 0.05) but down-regulated compared to the DN group (P ＜ 0.05). ③ VEGF staining in the DN group was darker compared to control group (P ＜ 0.01), while the staining in the irbesartan group was also darker compared to the control group (P ＜0.01), but the staining in the irbesartan group was lighter compared to the DN group (P ＜ 0.01). Flk-1 staining was similar to the VEGF. ④ VEGF and Flk-1 were positively correlated, with urine protein level, area and volume of renal glomerulus (P ＜0.05).CONCLUSION: VEGF and its Flk-1 receptor play important roles in DN pathogenesis. Over expressions may cause renal injury, but irbesartan (angiotensin Ⅱ receptor antagonist) has the protective effects on kidney through inhibiting abnormal expression of VEGF and Flk-1.
