长链非编码RNA 6030408B16RIK表达下调对腹膜间皮细胞上皮-间质转化的影响  

Effect of down-regulated expression of long noncoding RNA 6030408B16RIK on epithelial-mesenchymal transition in peritoneal mesenchymal cells

作　　者：籍文捷 王志奎[1] JI Wenjie;WANG Zhikui(Author Affiliation:Department of Nephrology,Linyi People's Hospital,Linyi,Shandong 276000,China)

机构地区：[1]临沂市人民医院肾脏内科,山东临沂276000

出　　处：《安徽医药》2022年第6期1118-1123,I0004,共7页Anhui Medical and Pharmaceutical Journal

基　　金：山东省自然科学基金项目(ZR2019MH126);山东省医药卫生科技发展计划项目(2018WS404)。

摘　　要：目的探讨长链非编码RNA(lncRNA)-6030408B16RIK在人腹膜间皮细胞上皮-间质转化(EMT)中的作用。方法于2019年3月至2020年6月,将36只大鼠以抽签法分为正常组6只、假手术组6只、尿毒症模型组24只,采用5/6肾切除法建立大鼠尿毒症模型,将尿毒症造模成功的大鼠再以抽签法分为:尿毒症组(不进行腹膜透析)、腹膜透析组(腹膜透析4周)、小干扰RNA阴性对照(si-NC)组(腹膜透析4周+空质粒)、si-6030408B16RIK组(腹膜透析4周+si-6030408B16RIK),每组6只。进行腹膜平衡试验检测大鼠超滤量和葡萄糖转运量,处死大鼠并对大鼠壁层腹膜进行苏木精-伊红(HE)染色、Masson染色和免疫组织化学观察腹膜组织结构变化及胶原纤维化情况,利用逆转录聚合酶链反应(RT-PCR)测定RNA的浓度和纯度,利用蛋白质印迹法检测各组上皮钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、成纤维细胞特异性蛋白-1(FSP-1)和波形蛋白的蛋白表达水平。结果与尿毒症组相比,腹膜透析组、si-NC组和si-6030408B16RIK组大鼠的超滤量明显降低(P<0.05),而葡萄糖转运量明显升高(P<0.05),大鼠腹膜厚度和Ⅲ型胶原蛋白(CollagenⅢ)、CD31的阳性表达量均显著增加(P<0.05),α-SMA、FSP-1和波形蛋白的表达均明显升高(P<0.05);相比于腹膜透析组,si-NC组大鼠的超滤量、葡萄糖转运量、腹膜厚度、CollagenⅢ、CD31及α-SMA、FSP-1、波形蛋白的表达均差异无统计学意义(P>0.05),而相比于si-NC组,si-6030408B16RIK组大鼠的超滤量[(5.45±0.57)mL比(4.23±0.43)mL]显著增加(P<0.05),葡萄糖转运量[(16.12±1.65)mmol/kg比(19.54±1.97)mmol/kg]显著减少(P<0.05),大鼠腹膜厚度、CollagenⅢ、CD31的表达量均显著减小(P<0.05),α-SMA、FSP-1和波形蛋白的表达[(0.65±0.06)、(1.29±0.13)、(1.39±0.14)比(1.24±0.12)、(1.72±0.19)、(1.99±0.21)]明显降低(P<0.05),E-cadherin表达[(0.96±0.10)比(0.38±0.04)]明显回升(P<0.05)。结论lncRNA 6030408B16RIK的�Objective To investigate the role of long noncoding RNA(lncRNA)6030408B16RIK in epithelial-mesenchymal transition(EMT)of human peritoneal mesenchymal cells.Methods From March 2019 to June 2020,36 rats were randomly assigned into normal group(n=6),sham operation group(n=6)and uremia group(n=24),and the uremia model was established by 5/6 nephrectomy method.The rats successfully modeled with uremia were then randomly subdivided into:Uremia group(no peritoneal dialysis),PD group(4-week peritoneal dialysis),small interfering RNA Negative Control(si-NC)group(4-week peritoneal dialysis+empty plasmid),si-6030408B16RIK group(4-week peritoneal dialysis+si-6030408B16RIK),6 rats for each subgroup.Ultrafiltration volume and glucose transport volumes of rats were measured by peritoneal equilibration test.The rats were sacrificed and the parietal peritoneum was stained by HE staining,and the changes of peritoneum tissue structure and collagen fibrosis were observed by Masson staining and immunohistochemistry.The concentration and purity of RNA were determined by reverse transcription polymerase chain reaction(RTPCR).Western blotting was used to detect the protein expression levels of E-cadherin,α-smooth muscle actin(α-SMA),fibroblast specific protein-1(FSP-1)and vimentin in each subgroup.Results Compared with Uremia group,the ultrafiltration volumes of PD group,si-NC group and si-6030408B16RIK group were significantly decreased(P<0.05),while glucose transport volumes were significantly increased(P<0.05).The peritoneal thickness and positive expressions of CollagenⅢand CD31 were significantly increased(P<0.05),and the expressions ofα-SMA,FSP-1 and vimentin were significantly increased(P<0.05).Compared with PD group,there were no significant differences in ultrafiltration volume,glucose transfer volume,peritoneal thickness,CollagenⅢ,CD31,α-SMA,FSP-1 and vimontin expressions in si-NC group.Compared with si-NC group,the ultrafiltration volume of si-6030408B16RIK group was significantly increased[(5.45±0.57)mL vs.(4.23±0.

关 键 词：上皮-间质转化 尿毒症 6030408B16RIK 腹膜透析 超滤衰竭 胶原Ⅲ型 大鼠 Sprague-Dawley 

分 类 号：R692.5[医药卫生—泌尿科学]