摘要
目的探讨intermedin（IMD）预处理对大鼠肾脏缺血再灌注损伤（IRI）修复再生过程中相关修复基因的影响。方法健康雄性Wistar大鼠144只，随机分为假手术组、IRI组、转染空质粒组和转染IMD组。动物右肾切除后，用超声微泡技术将质粒转染入左肾，1周后制作肾脏IRI模型。分别于再灌注后1d、2d、3d、4d、7d和14d留取肾组织标本。RT—PCR法检测肾组织Pax-2、ZO-1、Ncam、Wt-1和vimentin的mRNA表达变化；Western印迹法分析Pax-2、Wt-1和vimentin蛋白表达变化；ELISIA法检测ZO-1、Ncam的蛋白表达变化。结果（1）IRI组大鼠Pax-2、Zo—1、Ncam、Wt-1和vimentin的mRNA和蛋白的表达在IRI后1—3d增加，2d达到高峰，与假手术组相比差异有统计学意义（P〈0．05），4d后基本恢复。（2）在IRI后1～4d，转染IMD组上述的mRNA和蛋白表达与同时间点的假手术组、IRI组和转空质粒组相比差异均有统计学意义（均P〈0．05），其中2d达高峰。（3）在IRI后7d、14d，IRI组、转染空质粒组和转染IMD组上述指标的mRNA和蛋白表达和假手术组差异无统计学意义（均P〉．05）；在IRI后4d，IRI组和转空质粒组上述的表达较假手术组增高，但差异无统计学意义（P〉0．05）。（4）各时间点IRI组和转染空质粒组上述各指标差异均无统计学意义（P〉0．05）。（5）ELISA法检测的ZO-和Ncam蛋白表达变化趋势与RT—PCR及Western印迹法一致。结论IMD预处理可通过增加肾脏组织修复基因Pax-2、ZO-1、Ncam、Wt-1和vimentin的表达，从而对肾组织的修复和再生起到重要作用。
Objective To investigate the effects of intermedin （IMD） pretreatment on associated repairing genes of the rats with injured kidneys by ischemia reperfusion injury （IRI） （luring the repair and regeneration process. Methods A total of 144 healthy male Wistar rats were randomly divided into 4 gourps： sham opration group, IRI group, empty plasmid group （EP） and IMD group. After resection of right kidneys of the rats, plasmid was transfected into the left kidneys by using ultrasonic microbubble technology. After one week, the renal IRI models were programmed. The samples of renal tissues after 1 d, 2 d, 3 d, 4 d, 7 d and 14 d of reperfusion were harvested respectively, then the mRNA expressions of the Pax-2, ZO-1, Ncam, Wt-1 and vimentin in renal tissues were detected by RT-PCR; the protein expressions of Pax-2, Wt-1 and vimentin were analyzed by Western blotting and the protein expressions of ZO-1 and Ncam were measured by ELISIA. Results （1） Compared with the sham group, the mRNA and protein expression of Pax-2, ZO-1, Ncam, Wt-1 and vimentin of the rats in IRI group increased significantly from day 1 to day 3 （P〈0.05）, which peaked at day 2. After day 4, the above expressions in IR1 group returned to normal level. （2） In IMD group, the mRNA and protein expressions of Pax-2, ZO-1, Ncam, Wt-1 and vimentin were significantly higher than those in other 3 groups （P〈0.05） at the same time after IRI day 1 to day 4, with a maximum in day 2. （3） The above expressions in IRI group, EP group, IMD group had no significant differences compared with sham group after day 7 or day 14 respectively （P〉0.05）, and either between IRI group and EP group, though the expressions of the genes in IRI group and EP group increased compared with sham group after day 4. （4） The above expressions between IRI group and EP group also had no significant difference （P〉0.05）. （5） Changes of ZO-1 and Neam protein expression detected by ELISA were similar to those abore mRNA and protein expression by RT-PCR and Western blotting. Conclusion IMD pretreatment plays an important role in up-regulation of the expressions of associated repair genes during the process of repair and regeneration after renal ischemia reperfusion injury.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2011年第9期678-683,共6页
Chinese Journal of Nephrology
基金
国家自然科学基金面上项目（30971380）
山西省科技攻关项目（20080311061-6）
