CAJ | 학술논문

目的探讨幼鼠单侧输尿管完全梗阻(complete unilateral ureteral obstruction,CUUO)肾脏miRNA-101表达与肾间质纤维化进展的关系.方法选取30只SD雄性幼鼠,年龄(8±1)周,体重(180±20)g,数字随机分为sham组、CUUO 7 d组和14d组.分别于梗阻后7d和14d处死取其梗阻侧肾脏,将其切开分为均等的两部分,一部分用于RT-PCR,采用相对定量法检测不同梗阻时间段梗阻侧肾脏中miRNA-101表达.另一部分用于Western blot及免疫组织化学检测α-SMA、E-cadherin的相对定量及定性表达.结果免疫组织化学测定结果显示α-SMA及E-cadherin主要表达于肾间质,呈棕黄色,其中α-SMA和E-cadherin在CUUO 14 d组、UUO 7 d组及sham组显色依次递减和增强;组织学上可见CUUO 7 d组及14d组肾小管管腔增大,明显不同于sham组.RT-PCR结果显示miRNA-101相对表达量(fold difference) sham组是CUUO 14 d组的(14.50±5.12)倍,CUUO7d组是14d组的(3.45±2.27)倍,三组间差异有统计学意义(P＜0.05);Western blot显示梗阻侧肾脏α-SMA在CUUO 14 d组、CUUO 7 d和sham表达分别为1.22±0.48、0.50±0.06和0.21±0.03,与相应时间段miRNA-101表达成负相关(r=-0.74,P＜0.05).相反,E-cadherin在CUUO 14 d组、CUUO 7 d和sham表达分别为0.24±0.22、0.50±0.44和0.76±0.20.与相应时间段miRNA-101表达成正相关(r=0.88,P＜0.05).结论幼鼠完全梗阻肾脏miRNA-101表达随着梗阻时间的延长而下降,与反映肾脏间质纤维化严重程度的蛋白表达相关,提示miRNA-101的变化可能参与梗阻肾脏纤维化过程的进展.
목적 탐토유서단측수뇨관완전경조(complete unilateral ureteral obstruction,CUUO)신장miRNA-101표체여신간질섬유화진전적관계.방법 선취30지SD웅성유서,년령(8±1)주,체중(180±20)g,수자수궤분위sham조、CUUO 7 d조화14d조.분별우경조후7d화14d처사취기경조측신장,장기절개분위균등적량부분,일부분용우RT-PCR,채용상대정량법검측불동경조시간단경조측신장중miRNA-101표체.령일부분용우Western blot급면역조직화학검측α-SMA、E-cadherin적상대정량급정성표체.결과 면역조직화학측정결과현시α-SMA급E-cadherin주요표체우신간질,정종황색,기중α-SMA화E-cadherin재CUUO 14 d조、UUO 7 d조급sham조현색의차체감화증강;조직학상가견CUUO 7 d조급14d조신소관관강증대,명현불동우sham조.RT-PCR결과현시miRNA-101상대표체량(fold difference) sham조시CUUO 14 d조적(14.50±5.12)배,CUUO7d조시14d조적(3.45±2.27)배,삼조간차이유통계학의의(P＜0.05);Western blot현시경조측신장α-SMA재CUUO 14 d조、CUUO 7 d화sham표체분별위1.22±0.48、0.50±0.06화0.21±0.03,여상응시간단miRNA-101표체성부상관(r=-0.74,P＜0.05).상반,E-cadherin재CUUO 14 d조、CUUO 7 d화sham표체분별위0.24±0.22、0.50±0.44화0.76±0.20.여상응시간단miRNA-101표체성정상관(r=0.88,P＜0.05).결론 유서완전경조신장miRNA-101표체수착경조시간적연장이하강,여반영신장간질섬유화엄중정도적단백표체상관,제시miRNA-101적변화가능삼여경조신장섬유화과정적진전.
Objective To explore the correlation of miRNA-101 expression with the progression of renal interstitial fibrosis in young rats with complete unilateral ureteral obstruction (CUUO).Methods Thirty male Sprague Dawley rats (age 8 ± 1 weeks,weight 180 ± 20 g) were randomly divided into 3 groups of sham,CUUO 7 days (D7) and 14 days (D14).The kidney samples were collected at Days 7 and 14 respectively and used for detecting the expression of miRNA-101 by realtime polymerase chain reaction (RT-PCR) and α-SMA and E-cadherin protein by Western blot and immunohistochemistry.The correlations between their expressions were analyzed.Results Immunohistochemistry showed that mSMA and E-cadherin were predominantly expressed in renal interstitium.The expression of a-SMA gradually declined in all groups.However there was a gradually elevated expression of E-cadherin.The tubular lumen of D7 and D14 groups expanded more dramatically than that of sham group.RT-PCR results showed that the expression magnitude of MiRNA-101 in sham and D7 groups was (14.50 ± 5.12) and (3.45 ± 2.27) folds of D14 group respectively.Significant difference existed among 3 groups (P＜ 0.05).The expression level of α-SMA protein was 1.22 ± 0.48,0.50 ± 0.06 and 0.21 ± 0.03 in D14,D7 and sham respectively and there were negative correlations with miRNA-101 (r＝-0.74,P＜0.05).In contrast,the expression level of E-cadherin was 0.24 ± 0.22,0.50 ± 0.44 and 0.76 ±0.20 in D14,D7 and sham respectively and there were positive correlations with miRNA-101 (r =0.88,P＜0.05).Conclusions The renal expression of miRNA-101 decreases with obstructive duration and it is related with renal interstitial fibrosis proteins.And miRNA-101 may be significantly correlated with the progression of renal interstitial fibrosis.