Resolvin D1 Protects Lipopolysaccharide-induced Acute Kidney Injury by Down-regulating Nuclear Factor-kappa B Signal and Inhibiting Apoptosis

• Published in: Chinese medical journal Vol. 129; no. 9; pp. 1100 - 1107
• Main Authors: Zhao, Yu-Liang, Zhang, Ling, Yang, Ying-Ying, Tang, Yi, Zhou, Jiao-Jiao, Feng, Yu-Ying, Cui, Tian-Lei, Liu, Fang, Fu, Ping
• Format: Journal Article
• Language: English
• Published: China Wolters Kluwer - Medknow Publications 05.05.2016; Medknow Publications and Media Pvt. Ltd; Lippincott Williams & Wilkins Ovid Technologies; Division of Nephrology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China; Medknow Publications & Media Pvt Ltd; Wolters Kluwer
• Subjects: Acute Kidney Injury - chemically induced; Acute Kidney Injury - prevention & control; Acute Kidney Injury; Apoptosis; Lipopolysaccharide; Nuclear Factor-kappa B; Resolvin D1; Adaptor Proteins, Signal Transducing - analysis; Analysis; Animals; Apoptosis; Apoptosis - drug effects; Docosahexaenoic Acids - pharmacology; Down-Regulation; Kidney - drug effects; Kidney - pathology; Kidney diseases; Kidneys; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Male; Mice; Mice, Inbred BALB C; NF-kappa B - antagonists & inhibitors; Original; Risk factors; Rodents; Tumor Necrosis Factor-alpha - analysis; Tumor necrosis factor-TNF; 保护作用; 信号通路; 免疫组织化学染色; 凋亡诱导; 核因子κB; 肾小管上皮细胞; 肾损伤; 脂多糖; United States > US; Lipopolysaccharide; Nuclear Factor-kappa B; Acute Kidney Injury; Resolvin D1; Apoptosis
• ISSN: 0366-6999; 2542-5641
• DOI: 10.4103/0366-6999.180517

Background： Resolvin D1 （RvD1） is a newly found anti-inflammatory bioactive compound derived from polyunsaturated fatty acids. The current study aimed to explore the protective effect of RvD1 on lipopolysaccharide （LPS）-induced acute kidney injury （AKI） and its possible mechanism. Methods： Both in vivo and in vitro studies were conducted. Male BALB/c mice were randomly divided into control group （saline）, LPS group （LPS 5 mg/kg）, RvD1 group （RvD1 5 μg/kg ＋ LPS 5 mg/kg）, and blockage group （Boc-MLP 5 gg/kg ＋ RvD1 5 gg/kg ＋ LPS 5 mg/kg）. Boc-MLP is a RvD 1 receptor blocker. The mice were intraperitoneally injected with these drugs and recorded for general condition for 48 h, while the blood and kidneys were harvested at 2, 6, 12, 24, and 48 h time points, respectively （n = 6 in each group at each time point）. Human proximal tubule epithelial cells （HK-2） were randomly divided into control group （medium only）, LPS group （LPS 5 μg/ml）, RvD1 group （RvD1 10 ng/ml ＋ LPS 5 μg/ml）, and blockage group （Boc-MLP 10 ng/ml ＋ RvD1 10 ng/ml ＋ LPS 5 μg/ml）. The cells were harvested for RNA at 2, 4, 6, 12, and 24 h time points, respectively （n = 6 in each group at each time point）. Blood creatinine was tested by using an Abbott i-STAT portable blood gas analyzer. Tumor necrosis factor-α （TNF-α level was detected by EL1SA. Kidney pathology was observed under hematoxylin and eosin （HE） staining and transmission electron microscope （TEM）. We hired immune-histological staining, Western blotting, and fluorescence quantitative polymerase chain reaction to detect the expression of RvD1 receptor ALX, nuclear factor-kappa B （NF-KB） signaling pathway as well as caspase-3. Kidney apoptosis was evaluated by TUNEL staining. Results： RvD 1 receptor ALX was detected on renal tubular epithelials. Kaplan-Meier analysis indicated that RvD 1 improved 48 h animal survival （80%） compared with LPS group （40%） and RvDI blockage group （60%）, while RvD1 also ameliorated kidney pathological injury in HE staining and TEM scan. After LPS stimulation, the mRNA expression of toll-like receptor 4, myeloid differentiation factor 88, and TNF-α in both mice kidneys and HK-2 cells were all up-regulated, while RvDI substantially inhibited the up-regulation of these genes. Western blotting showed that the phosphorylated-IKB/IKB ratio in LPS group was significantly higher than that in the control group, which was inhibited in the RvD1 group. RvD1 could inhibit the up-regulation of cleaved-caspase-3 protein stimulated by LPS, which was prohibited in RvD 1 blockage group. RvD 1 group also had a lower proportion of apoptotic nuclei in mice kidney by TUNEL staining compared with LPS group. Conclusion： In LPS-induced AKI, RvD1 could decrease TNF-α level, ameliorate kidney pathological injury, protect kidney function, and improve animal survival by down-regulating NF-KB inflammatory signal as well as inhibiting renal cell apoptosis.