Prevention of renal ischemia-reperfusion injury by short hairpin RNA of endothelin A receptor in a rat model.
Endothelin A receptor (ETaR) is a key molecule involved in a variety of biological events such as vessel contraction and inflammatory response in ischemia-reperfusion (I/R) injury. RNA interference using short hairpin RNA (shRNA) is a powerful tool to silence gene expression. Here, the effect of ETaR shRNA on I/R injury in rats was studied. A more effective shRNA sequence out of two constructed into plasmid vectors was selected using the A-10 cell line, and was then applied to a rat model. Twenty-eight male Sprague-Dawley rats were randomized into four groups: Sham, shRNA, vector and phosphate-buffered saline (PBS). Renal I/R injury was induced by clamping the left renal pedicle for one hour followed by reperfusion for 24 h. ETaR shRNA (100 μg) plasmid was administered by renal vein injection 48 h before clamping. The expression of both ETaR mRNA and protein was lowered by ETaR shRNA treatment compared with that in the vector and PBS groups; serum creatinine and blood urea nitrogen were significantly decreased; the semi-quantitative score of renal structural damage was improved; the mRNA level of endothelin 1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), macrophage inflammatory protein 2 (MIP-2) and monocyte chemoattractant protein 1 (MCP-1) was reduced, but nitric oxide (NO) production in kidney tissues was increased (P < 0.05). In conclusion, ETaR shRNA partially silenced ETaR expression in I/R injury kidneys, reduced the mRNA level of ET-1, inflammatory mediators including TNF-α, IL-6, MIP-2 and MCP-1, increased NO production, and ultimately improved renal function and structure.
Shanghai Key laboratory of Organ Transplantation, Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, P R China.
Yichen
Y
Jia
Zitong
Z
Zhao
Ming
M
Xu
Tian
T
Zhao
Yongyin
Y
Qiu
Yitreen
Y
Ooi
Bin
B
Yang
Ruiming
R
Rong
Tongyu
T
Zhu
D000818:Animals; D001688:Biological Products / Q000008:administration & dosage* / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D065130:Endothelin A Receptor Antagonists; D020869:Gene Expression Profiling; D007275:Injections, Intravenous; D007668:Kidney / Q000473:pathology; D008297:Male; D010957:Plasmids / Q000008:administration & dosage; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D044022:Receptor, Endothelin A / Q000235:genetics; D015427:Reperfusion Injury / Q000517:prevention & control*
D001688:Biological Products; D016207:Cytokines; D065130:Endothelin A Receptor Antagonists; D034741:RNA, Small Interfering; D044022:Receptor, Endothelin A