摘要
目的：探讨 miRNA-101、ZESTE 基因增强子同源物2（EZH2）及转化生长因子（TGF）-β1在幼鼠单侧输尿管完全梗阻（CUUO）肾脏中的表达和意义。方法选取30只 SD 雄性幼鼠，周龄（6±1）周，体质量（150±10）g，采用随机数字表法分为 sham 组、CUUO 7 d 组、CUUO 14 d 组，每组10只。分别于梗阻后7 d 及14 d 处死取其梗阻侧肾脏，横断面切开分为均等的上下段2部分，上段部分用于 Western blot 检测肾脏 TGF-β1及 EZH2的表达；下段部分再纵切为均等的2部分，一部分用于实时荧光定量-聚合酶链反应（RT-PCR）检测不同梗阻时间段肾脏 miRNA-101的表达，另一部分用于免疫组织化学及 HE 染色检测肾脏 TGF-β1及 EZH2的定性表达。结果 RT-PCR 结果显示 miRNA-101相对表达量：sham 组是 CUUO 14 d 组的（12.69±1.60）倍， CUUO 7 d组是14 d 组的（3.74±1.24）倍，3组间差异有统计学意义（P ﹤0.05）；Western blot 显示 TGF-β1相对表达量在 CUUO 14 d、7 d 及 sham 组分别为1.14±0.12、1.04±0.14和0.76±0.18；EZH2相对表达量在 CUUO 14 d、7 d 及 sham 组分别为1.04±0.04、0.89±0.03和0.73±0.02；miRNA-101与 EZH2表达呈负相关（r ＝-0.92，P ﹤0.05），与 TGF-β1表达呈负相关（r ＝-0.63，P ﹤0.05），EZH2与 TGF-β1表达呈正相关（r ＝0.67， P ﹤0.05）。不同梗阻时间肾脏 miRNA-101、EZH2及 TGF-β1的表达均具有明显相关性。结论随着梗阻时间的延长，miRNA-101表达下降，EZH2及 TGF-β1表达上升，提示 miRNA-101可能通过调节幼鼠肾脏 EZH2表达影响肾间质纤维化进展。
Objective To explore the expressions and significance of miRNA - 101,enhancer of ZESTE homo-log 2（EZH2）and transforming growth factor（TGF）- β1 in the kidneys with complete uniateral ureteral obstruction （CUUO）. Methods Thirty male SD rats,（6 ± 1）weeks old,weighted（150 ± 10）g,were divided into sham group, 7 - day group with CUUO and 14 - day group with CUUO by using random number table method,10 rats in each group. The obstructed kidney samples were collected in 7 and 14 days,respectively,for detecting the expression of miRNA -101 by real time - polymerase chain reaction（RT - PCR）and TGF - β1 and EZH2 protein by Western blot,immuno-histochemistry and hematoxylin - eosin staining. Their correlated expressions were analyzed. Results RT - PCR results showed that the expressions of miRNA - 101 in sham group and 7 - day group with CUUO were（12. 69 ± 1. 60）times and（3. 74 ± 1. 24）times which were higher than those of 14 - day group with CUUO,respectively,there was a signifi-cant difference among these 3 groups（P ﹤ 0. 05）. The expressions of TGF - β1 and EZH2 proteins were 1. 14 ± 0. 12, 1. 04 ± 0. 14,0. 76 ± 0. 18 and 1. 04 ± 0. 04,0. 89 ± 0. 03,0. 73 ± 0. 02 in 14 - day group with CUUO,7 - day group with CUUO and sham group,respectively. There was a negative correlation between miRNA - 101 with EZH2（ r =- 0. 92,P ﹤ 0. 05）,and negative correlation with TGF - β1（r = - 0. 63,P ﹤ 0. 05）,and positive correlation between EZH2 and TGF - β1（r = 0. 67,P ﹤ 0. 05）;the expressions of miRNA - 101,EZH2 and TGF - β1 were associated with each other in obstruction renal in different time periods. Conclusions With the extension of obstruction time, miRNA - 101 expression decreased,EZH2 and TGF - β1 expression increased,evidently,which indicates that the de-velopment of renal interstitial fibrosis may be affected through regulating EZH2 expression of renal obstruction through miRNA - 101 in the young rats.
出处
《中华实用儿科临床杂志》
CSCD
北大核心
2015年第21期1656-1659,共4页
Chinese Journal of Applied Clinical Pediatrics
基金
基金项目：国家自然科学基金（81370869）
河南省卫计委科研项目（201303020）
郑州市科技局创新人才（131PLJRC657）
