【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。更多还原

【Abstract】 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 in- traperitoneal injection with 0. 9% normal saline; ( 3) hypertonic dialysate group( n = 8) : daily intraperitoneal injection with 4. 25% peritoneal dialysis solution. Rats were sacrificed after four weeks. A 2 - hour peritoneal equilibration test ( PET) was performed. Per- itoneal 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 ultrafiltration ( UF) and glucose reabsorption ( D2 /D0 ) were significantly lower and dialysate - to - plasma urea ratio( D/Purea) was significantly higher in the hypertonic dialysate group,which in- dicated peritoneal was high solute transport,and peritoneal function was decreased; The hypertonic dialysate group had more perito- neal vessels and inflammatory cells,The results of microarray analysis showed that miR - 100、miR - 152 and miR - 497 were down- regulated in the hypertonic dialysate group,miR - 192 and miR - 194 were downregulated in the hypertonic 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 miR- NAs 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.更多还原