Protective effect of ETaR siRNA on renal ischemia-reperfusion injury in rats by changing the immuno-microenvironment of kidney

Chineae Journal of Organ Transplantation Pub Date : 2019-02-20 DOI:10.3760/CMA.J.ISSN.0254-1785.2019.02.002

Yichen Jia, Long Zheng, Long Li, Jiawei Li, Ming Xu, T. Zhu

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Abstract

Objective To explore the protective effect of ETaR siRNA on renal ischemia reperfusion injury (IRI) by changing the immuno-microenvironment in rats. Methods A total of 40 male Sprague-Dawley (SD) rats were randomized into four groups of sham, IR, negative siRNA and ETaR siRNA. A renal IRI model was generated by clamping left renal artery. ETaR siRNA was delivered into kidney through renal vein by a retrograde 'hydrodynamic’ injection. Blood samples were collected for detecting renal function and kidney tissue harvested for Hematoxylin & Eosin (HE) staining, TdT-mediated dUTP Nick-End Labeling (TUNEL) staining, polymerase chain reaction (PCR) and Western blot at 48 h post-reperfusion. Results Serum creatinine, blood urea nitrogen and renal apoptotic cells increased and renal tissue was injured after IR. The changes were inhibited by ETaR siRNA. PCR showed that ETaR siRNA treatment significantly down-regulated the expressions of inflammatory factors TNF-α, IFN-γ and IL-6 and transcription factor NF-κB induced by IR. Conclusions ETaR siRNA can effectively improve the immuno-microenvironment and thereby alleviate renal ischemia reperfusion injury. Key words: Rats; Kidney; Endothelin receptor; Small interfering RNA; Immuno-microenvironment; Reperfusion injury

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ETaR siRNA通过改变肾脏免疫微环境对大鼠肾缺血再灌注损伤的保护作用

目的探讨ETaR siRNA通过改变免疫微环境对大鼠肾缺血再灌注损伤(IRI)的保护作用。方法选取雄性SD大鼠40只，随机分为sham组、IR组、阴性siRNA组和ETaR siRNA组。通过夹持左肾动脉建立肾IRI模型。ETaR siRNA通过肾静脉逆行“流体动力学”注射进入肾脏。再灌注后48 h，采集血样检测肾功能，采集肾组织进行苏木精&伊红(HE)染色、tdt介导的dUTP镍端标记(TUNEL)染色、聚合酶链反应(PCR)和Western blot。结果IR后大鼠血清肌酐、血尿素氮及肾细胞凋亡增加，肾组织损伤。这些变化被ETaR siRNA抑制。PCR结果显示，ETaR siRNA处理显著下调IR诱导的炎症因子TNF-α、IFN-γ、IL-6及转录因子NF-κB的表达。结论ETaR siRNA可有效改善免疫微环境，从而减轻肾缺血再灌注损伤。关键词:大鼠;肾脏;内皮素受体;小干扰RNA;Immuno-microenvironment;再灌注损伤

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Chineae Journal of Organ Transplantation

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