增殖细胞核抗原和caspase-3在单侧输尿管梗阻大鼠模型中的表达及其意义  

Expression and Significance of Proliferating Cell Nuclear Antigen and caspase-3 Expression in Rat Model of Unilateral Ureteral Obstruction

作　　者：李里[1] 宫亮[2] 吴玉斌[3] 

机构地区：[1]辽宁医学院附属第一医院儿科,辽宁省锦州市121000 [2]辽宁医学院附属第一医院耳鼻喉科,辽宁省锦州市121000 [3]中国医科大学附属盛京医院小儿肾脏风湿免疫科

出　　处：《中国全科医学》2012年第30期3489-3493,共5页Chinese General Practice

基　　金：沈阳市科学技术计划项目(F10-205-1-29);辽宁医学院第一临床医院青年科技启动基金项目(FY2011-05);辽宁医学院青年科技启动基金项目(F2011Z005)

摘　　要：目的探讨增殖细胞核抗原(PCNA)和caspase-3在单侧输尿管梗阻大鼠模型中的表达及其意义。方法 2011年5—12月选取清洁级Wistar大鼠48只进行随机对照试验,采用随机数字表法分为两组,假手术组(sham组)和模型组(UUO组)各24只,sham组分离输尿管不结扎,UUO组行左侧输尿管结扎术。每组术后3、7、14 d分为3组,每组8只,生化分析仪检测大鼠尿β2微球蛋白和血肌酐水平;苏木素-伊红(HE)染色,光镜观察肾脏病理形态改变;免疫组织化学染色(SABC)法、蛋白印迹(Western blot)法检测PCNA和caspase-3的表达。采用SPSS 13.0统计软件进行统计学处理。结果术后3、7 d两组尿β2微球蛋白水平比较,差异均无统计学意义(P>0.05);术后14 d两组尿β2微球蛋白水平比较,差异有统计学意义(P<0.05)。术后3、7、14 d两组血肌酐水平比较,差异均无统计学意义(P>0.05)。HE染色显示,sham组大鼠肾脏肾小管排列紧密、整齐;UUO组术后3 d,可见肾小管肿胀,部分肾小管轻度扩张,小管间质区域出现炎性浸润;术后7 d肾小管壁扩张,肾间质水肿;术后14 d肾小管结构已经完全被破坏,管腔塌陷。SABC法结果显示,两组术后3、7、14 d,肾小管内PCNA阳性细胞百分率比较,差异无统计学意义(P>0.05);肾间质内PCNA阳性细胞百分率比较,差异均有统计学意义(P<0.05)。两组术后3、7、14 d,肾小管内caspase-3阳性细胞百分率比较,差异均有统计学意义(P<0.05);肾间质内caspase-3阳性细胞百分率比较,差异均有统计学意义(P<0.05)。Western blot法结果显示,sham组肾皮质中有微量PCNA、caspase-3表达;UUO组术后PCNA、caspase-3表达增加,梗阻时间延长表达增加。结论 PCNA表达增加导致肾间质细胞过度增殖,参与梗阻性肾病病理过程。caspase-3活性明显增强,加速细胞凋亡,在梗阻性肾病中对起着重要作用。细胞的增殖与凋亡参与UUO大鼠模型肾纤维化过程,PCNA和caspase 3是这一过程的两个Objective To investigate the expression and significance of proliferating cell nuclear antigen （PCNA） and Caspase - 3 in the rat model of unilateral ureteral obstruction. Methods 48 wistar rats of clean grade were randomly divided into sham operation group （ sham group） and experimental model group （ UUO group） with each group 24 rats. Rats in the sham group were given ureter separation without ureter ligation and rats in the UUO group were given left ureter ligation. The rats were sacri- riced three days, seven clays and 14 days after operation and were divided into six groups with each group eight rats. β2- microglobulin and blood serum creatinine were detected by biochemistry analyzer. Light microscope was used to observe pathomorphological changes after HE staining. SABC method and western blot method were used to detect the protein expressions of PCNA and easpase -3. SPSS 13.0 statistical software was used to make statistics analysis. Results The levels of β2 - microglobulin three clays and seven days after operation between the two groups showed no statistically significant difference （ P 〉 0. 05 ） . The levels of β2 - mieroglobulin 14 days after operation between the two groups showed statistically significant difference （ P 〈 0. 05 ） . The levels of serum creatinine three days, seven days and 14 days after operation between the two groups all showed no statistically significant difference （ P 〉 0. 05 ） . HE staining showed that kidney tubules were closely and regularly arranged in the sham group. In UUO group, kidney tubules swelled, part of the tubules expanded and tubulointerstitial areas showed inflammatory infil- tration three days after operation ; tubules wall expanded and tubulointerstitial areas showed edema seven days after operation ; the structure of tubules was completely destroyed and the lumen collapsed 14 days after operation. The results of SABC method showed that the percentage of positive cells of PCNA protein expression in tubules between the two groups

关 键 词：输尿管梗阻 大鼠 半胱氨酸天冬氨酸蛋白酶3 增殖细胞核抗原 

分 类 号：R693[医药卫生—泌尿科学]