Du Hongyin, Sheng Mingwei, Wu Li, Zhang Yamin, Shi Dongjing, Weng Yiqi, Xu Rubin, Yu Wenli
1 Department of Anesthesiology, Tianjin First Center Hospital, Tianjin, China. 2 Department of Pathology and Pathophysiology, Tianjin Medical University, Tianjin, China. 3 Department of Hepatobiliary Surgery, Tianjin First Center Hospital, Tianjin, China.
Transplantation. 2016 Mar;100(3):563-70. doi: 10.1097/TP.0000000000001052.
Acute kidney injury (AKI) impacts the survival of liver transplant recipients severely. To date, the related mechanism and effective therapy have not been rigorously explored. The present study aimed to explore the role of p53-mediated autophagy in the protective effect of hydrogen-rich saline (HRS) on AKI after orthotropic liver transplantation (OLT).
Adult male Sprague-Dawley rats were randomly allocated into four groups: sham, OLT, OLT with HRS (6 ml/kg) pretreatment (HS), OLT with HRS and chloroquine pretreatment (60 mg/kg) group (CQ). All the samples were collected 6 hours after reperfusion. The renal function and oxidative stress level were measured by biochemical and histopathologic examinations. The formation of autophagosome was observed by transmission electron microscopy. The apoptotic rate was determined by terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick-end labeling analysis. The expression of caspase-3, cytochrome c, p53, damage-regulated autophagy modulator, Becline-1, microtubule-associated protein light 3-II, p62, lysosome-associated membrane protein-2, and the phosphorylation of p53 were assayed by western blot assay.
Compared with the OLT group, HRS dramatically attenuated the histopathologic damage, restored the renal function, and decreased the oxidative stress level. Simultaneously, HRS significantly ameliorated apoptosis by decreasing the apoptotic rate and inhibiting the expression of caspase-3 and cytochrome c in rats subjected to OLT. The expression of Becline-1 and microtubule-associated protein light 3-II were upregulated with the inhibition of p62 and lysosome-associated membrane protein-2. The inhibition of autophagy by chloroquine counteracted the renoprotective effects of HRS.
HRS is able to protect against AKI after liver transplantation partly by reducing apoptosis, which is possibly involved in the modulation of p53-mediated autophagy.
急性肾损伤（AKI）严重影响肝移植受者的生存。迄今为止，相关机制和有效治疗方法尚未得到严格探索。本研究旨在探讨p53介导的自噬在富氢盐水（HRS）对原位肝移植（OLT）后AKI的保护作用中的作用。
将成年雄性Sprague-Dawley大鼠随机分为四组：假手术组、OLT组、OLT+HRS（6 ml/kg）预处理组（HS组）、OLT+HRS+氯喹（60 mg/kg）预处理组（CQ组）。再灌注6小时后收集所有样本。通过生化和组织病理学检查测量肾功能和氧化应激水平。通过透射电子显微镜观察自噬体的形成。通过末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记分析确定凋亡率。通过蛋白质印迹法检测半胱天冬酶-3、细胞色素c、p53、损伤调节自噬调节剂、Beclin-1、微管相关蛋白轻链3-II、p62、溶酶体相关膜蛋白-2的表达以及p53的磷酸化。
与OLT组相比，HRS显著减轻了组织病理学损伤，恢复了肾功能，并降低了氧化应激水平。同时，HRS通过降低凋亡率和抑制OLT大鼠中半胱天冬酶-3和细胞色素c的表达，显著改善了凋亡。Beclin-1和微管相关蛋白轻链3-II的表达上调，同时p62和溶酶体相关膜蛋白-2受到抑制。氯喹抑制自噬抵消了HRS的肾脏保护作用。
HRS能够部分通过减少凋亡来预防肝移植后的AKI，这可能与p53介导的自噬调节有关。
