大鼠腹膜纤维化相关miRNA的初步筛选  

The Preliminary Screening of miRNA Related with Rat Peritoneal Fibrosis

作　　者：吴旭[1] 林凡[1] 黄朝兴[1] 章慧娣[1] 尤小寒[1] 张周沧[1] 邵蓉蓉[1] 

机构地区：[1]温州医学院附属第一医院肾内科,温州325000

出　　处：《中国中西医结合肾病杂志》2013年第3期203-206,I0002,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：浙江省医药卫生科技计划项目(No.2010KYA135)

摘　　要：目的:初步筛选参与腹膜透析相关腹膜纤维化的miRNA。方法:将SD雄性大鼠24只[体重(150±10)g]随机平均分为3组:正常对照组;生理盐水组,每日腹腔注射0.9%生理盐水20ml;高糖组,每日腹腔内注射4.25%含糖透析液20ml。4周后,进行腹膜平衡试验(PET)。取大鼠腹膜组织行光镜检查,基因芯片分析检测各组大鼠腹膜组织miRNA。选取芯片分析显著差异表达的miRNAs,进行real-timePCR验证。结果:高糖组大鼠的超滤量和透出液葡萄糖浓度/初始腹透液葡萄糖浓度(D2/D0)明显减少,透析液尿素浓度/血浆尿素浓度(D/Purea)明显增加,提示高糖组大鼠腹膜呈高转运、腹膜功能下降。高糖组腹膜间皮下基质增厚、血管增生及炎症细胞浸润明显增加。基因芯片分析提示:在高糖组中,miR-100、miR-152与miR-497表达呈明显下调。在高糖组与生理盐水组中,miR-192、miR-194表达均呈显著下调。real-timePCR结果提示miR-100、miR-152、miR-497、miR-192、miR-194在高糖组中均呈低表达,与芯片分析结果基本一致。结论:在高浓度腹膜透析液诱导的大鼠腹膜纤维化模型中,多条miRNAs呈一致性差异表达。提示miRNA可能参与大鼠腹膜纤维化。Objective：To preliminarily screen miRNA of peritoneal fibrosis associated with peritoneal dialysis. Methods： Twenty - four SD rats were randomly divided into three groups ： （ 1 ） Control group （ n = 8 ） ; （2） Normal saline group （ n = 8 ） ： daily intraperitoneal injection with 0.9% normal saline; （ 3 ） hypertonic dialysate group （ n = 8） ： daily intraperitoneal injec/ion with 4.25% peritoneal dialysis solution. Rats were sacrificed after four weeks. A 2 - hour peritoneal equilibration test （PET） was performed. Peritoneal membrane histology was evaluated by light microscopy, and the expression of miRNAs were estimated by using microarray anal- ysis. miRNAs expression was proved by real -time PCR. Results ：The ultrafihration （UF） and glucose reabsorption （D2/D0 ） were significantly lower and dialysate -to -plasma urea ratio（D/Pure） was significantly higher in the hypertonic dialysate group, which indicated peritoneal was high solute transport, and peritoneal function was decreased; The hypertonic dialysate group had more peritoneal vessels and inflammatory cells ,The results of microarray analysis showed that miR -100 ,miR- 152 and miR- 497 were downregulated in the hypertonic dialysate group, miR - 192 and miR - 194 were downregulated in the hypertanic dialysate group and the normal saline group, The results of real - time PCR showed that miR - 100.miR - 152,miR -497.miR - 192 and miR - 194 were downregulated in the hypertonic dialysate group, which is consistent with microarray analysis. Conclusion：Differences in several miRNAs were expressed consistently in the peritoneal fibrosis rat model induced by 4.25% peritoneal dialysis solution, which indicated that miRNA might be closely correlated with the peritoneal fibrosis.

关 键 词：腹膜 纤维化 MIRNA 腹膜透析 

分 类 号：R692.5[医药卫生—泌尿科学]