Chen Jing, Zhang Han, Hu Jiachang, Gu Yulu, Shen Ziyan, Xu Linghan, Jia Xueqi, Zhang Xiaoyan, Ding Xiaoqiang
Department of Nephrology, Zhongshan Hospital, Fudan UniversityShanghai, China.
Kidney and Dialysis Institute of ShanghaiShanghai, China.
Front Pharmacol. 2017 Aug 11;8:499. doi: 10.3389/fphar.2017.00499. eCollection 2017.
Acute kidney injury (AKI) is a prominent risk factor for the development of chronic kidney disease (CKD). To date, the related mechanism and effective therapy have not been rigorously explored. The present study aims to investigate the reno-protection of hydrogen-rich saline (HRS) against ischemia/reperfusion (IR)-induced AKI. Adult male C57 mice were randomly allocated into three groups: Sham, IR, IR+HRS. Renal IR injury model was generated via 35 min occlusion of bilateral kidney pedicles, and then, mice were administered with different treatments intraperitoneally in various groups. After 14- or 28-day treatment, mice were perfused and the kidneys were collected following reperfusion. Many proteins were detected by western blots, including renal fibrotic proteins [a-smooth muscle actin (a-SMA), collagen I (Col I)], Klotho, the methylation of Klotho, damage-regulated autophagy modulator (Beclin-1), and microtubule-associated protein light 3-II (LC3-II). Finally, the levels of serum blood urea nitrogen (BUN) and creatinine (Cr) were measured to investigate the renal function. Histological data showed that the HRS treatment significantly decreased the fibrosis in renal tissues when compared with the IR group, and both of BUN and Cr were lower in the HRS group than IR group (8.9 ± 0.6 vs. 9.9 ± 0.1 mmol/l, 51 ± 6.5 vs. 60 ± 5.8 μmol/l) ( < 0.05). The expression of fibrotic markers, a-SMA and Col I, showed a robust increase in IR injury models than the Sham group, which was consistent with the result of Trichrome staining. However, the levels of a-SMA and Col I expression were sharply decreased in the IR+HRS group ( < 0.05). IR injury also enhanced LC3-II and Beclin-1 expression, but decreased Klotho level. The Klotho level was alleviated by HRS, but LC3-II and Beclin-1 were starkly enhanced in HRS group ( < 0.05). HRS showed a protective effect in the prevention of renal injury and could inhibit renal fibrosis after IR injury in mice. This role of HRS might be exerted via retaining Klotho expression and activating autophagy in the kidney.
急性肾损伤（AKI）是慢性肾脏病（CKD）发生的一个重要危险因素。迄今为止，相关机制和有效治疗方法尚未得到深入研究。本研究旨在探讨富氢盐水（HRS）对缺血/再灌注（IR）诱导的AKI的肾脏保护作用。成年雄性C57小鼠被随机分为三组：假手术组（Sham）、IR组、IR+HRS组。通过夹闭双侧肾蒂35分钟建立肾脏IR损伤模型，然后，对各组小鼠进行不同的腹腔内给药处理。治疗14天或28天后，对小鼠进行灌注，并在再灌注后收集肾脏。通过蛋白质印迹法检测了多种蛋白质，包括肾纤维化蛋白[α-平滑肌肌动蛋白（α-SMA）、I型胶原（Col I）]、Klotho、Klotho的甲基化、损伤调节自噬调节剂（Beclin-1）和微管相关蛋白轻链3-II（LC3-II）。最后，检测血清尿素氮（BUN）和肌酐（Cr）水平以评估肾功能。组织学数据显示，与IR组相比，HRS治疗显著降低了肾组织纤维化，且HRS组的BUN和Cr均低于IR组（8.9±0.6 vs. 9.9±0.1 mmol/l，51±6.5 vs. 60±5.8 μmol/l）（P<0.05）。纤维化标志物α-SMA和Col I的表达在IR损伤模型中比假手术组显著增加，这与三色染色结果一致。然而，在IR+HRS组中，α-SMA和Col I的表达水平急剧下降（P<0.05）。IR损伤还增强了LC3-II和Beclin-1的表达，但降低了Klotho水平。HRS可减轻Klotho水平降低，但HRS组中LC3-II和Beclin-1明显增强（P<0.05）。HRS对预防小鼠IR损伤后的肾损伤具有保护作用，并可抑制肾纤维化。HRS的这一作用可能是通过维持肾脏中Klotho的表达和激活自噬来实现的。
