Liu Nana, Chen Jing, Gao Dongmei, Li Wenhua, Zheng Di
Institute of Cardiovascular Disease Research, Xuzhou Medical University, Xuzhou, 221002, Jiangsu, China.
Department of Echocardiography, The First People's Hospital of Huaian, Huaian, 223001, Jiangsu, China.
Int Urol Nephrol. 2018 Jun;50(6):1171-1180. doi: 10.1007/s11255-018-1788-y. Epub 2018 Jan 24.
The study was processed to investigate the effect of astaxanthin (AST; 3,3-dihydroxybeta, beta-carotene-4,4-dione) on the acute kidney injury induced by iohexol and the relationship with SIRT1/FOXO3a signal pathway.
Thirty male Sprague Dawley rats were randomly divided into five groups as follows: control group (CON; olive oil only), contrast media group, astaxanthin control group (100 mg/kg), low astaxanthin dose group (LAG, 50 mg/kg) and high astaxanthin dose group (HAG, 100 mg/kg). As followed, serum creatinine (SCr), blood urea nitrogen (BUN), the oxidative stress markers and apoptosis-related proteins were detected. Human proximal tubular epithelial cells (HK-2) were cultured in DMEM/F12 medium in vitro and then randomly divided into appropriate experimental groups: normal group (N), dimethyl sulfoxide (DMSO), iohexol group (I), iohexol + (1.0, 10.0 μmol/L) astaxanthin group (I + LAST; I + HAST), iohexol + SIRT1 inhibitors (nicotinamide) group (NA) and iohexol + si-RNA FOXO3a group (si-RNA FOXO3a); when cultured for 24 h, cell proliferation ability was tested by cell counting kit (CCK-8), reactive oxygen species (ROS) were detected by flow cytometry and the expression of SIRT1 and FOXO3a were observed using western blot.
At the end of the experiment, the levels of SCr, BUN and malondialdehyde (MDA) were all increased in the CM group. The LAG and HAG reduce superoxide anion (SOD) activity, catalase (CAT) activity, glutathione peroxidase (GPx) activity and glutathione (GSH) content, as well as SCr and BUN level. Moreover, apoptosis-associated proteins, caspase 3 p17, bax and bcl-2 were upregulated. In HK-2 cells, after adding iohexol, proliferation and intracellular ROS levels were significantly increased. Using astaxanthin in advance after the intervention, the result is opposite. SIRTl inhibitors NA can reduce the expression of SIRTl and decrease the expression of FOXO3a protein. Si-RNA FOXO3a reduces the expression of FOXO3a but had no significant effect on the expression of SIRT1.
Our study demonstrates that the intervention of astaxanthin could attenuate the oxidative stress and apoptosis in contrast-induced acute kidney injury (CI-AKI), and the SIRT1/FOXO3a pathway participates in the contrast-induced apoptosis of HK-2 cells. Finally, astaxanthin reduces CI-AKI by suppression of apoptosis, which may be through inhibition of SIRT1/FOXO3a pathways.
本研究旨在探讨虾青素（AST；3,3 - 二羟基-β，β - 胡萝卜素 - 4,4 - 二酮）对碘海醇诱导的急性肾损伤的影响及其与SIRT1/FOXO3a信号通路的关系。
将30只雄性Sprague Dawley大鼠随机分为五组：对照组（CON；仅给予橄榄油）、造影剂组、虾青素对照组（100mg/kg）、低剂量虾青素组（LAG，50mg/kg）和高剂量虾青素组（HAG，100mg/kg）。随后，检测血清肌酐（SCr）、血尿素氮（BUN）、氧化应激标志物和凋亡相关蛋白。人近端肾小管上皮细胞（HK - 2）在体外DMEM/F12培养基中培养，然后随机分为适当的实验组：正常组（N）、二甲基亚砜（DMSO）组、碘海醇组（I）、碘海醇 +（1.0、10.0μmol/L）虾青素组（I + LAST；I + HAST）、碘海醇 + SIRT1抑制剂（烟酰胺）组（NA）和碘海醇 + si - RNA FOXO3a组（si - RNA FOXO3a）；培养24小时后，用细胞计数试剂盒（CCK - 8）检测细胞增殖能力，通过流式细胞术检测活性氧（ROS），并使用蛋白质免疫印迹法观察SIRT1和FOXO3a的表达。
实验结束时，造影剂组的SCr、BUN和丙二醛（MDA）水平均升高。LAG和HAG组降低了超氧化物歧化酶（SOD）活性、过氧化氢酶（CAT）活性、谷胱甘肽过氧化物酶（GPx）活性和谷胱甘肽（GSH）含量，以及SCr和BUN水平。此外，凋亡相关蛋白caspase 3 p17、bax和bcl - 2上调。在HK - 2细胞中，加入碘海醇后，细胞增殖和细胞内ROS水平显著升高。预先使用虾青素干预后，结果相反。SIRT1抑制剂NA可降低SIRT1的表达并减少FOXO3a蛋白的表达。Si - RNA FOXO3a降低了FOXO3a的表达，但对SIRT1的表达无显著影响。
我们的研究表明，虾青素干预可减轻造影剂诱导的急性肾损伤（CI - AKI）中的氧化应激和凋亡，且SIRT1/FOXO3a通路参与了碘海醇诱导的HK - 2细胞凋亡。最后，虾青素通过抑制凋亡减轻CI - AKI，这可能是通过抑制SIRT1/FOXO3a通路实现的。
