摘要/Abstract
摘要： 目的 ·探讨白细胞介素 -27（interleukin-27，IL-27）在顺铂所致急性肾损伤（ acute kidney injury，AKI）中的表达及在 AKI中对细胞调亡的抑制作用。方法 ·将 C57BL/6小鼠随机分为对照组、顺铂组，于注射顺铂后 24、48、72 h处死小鼠；取血清样本，检测血尿素氮和血清肌酐水平；实时荧光定量 PCR检测肾脏组织中 IL-27亚单位 p28 和 EBI3 在 mRNA水平的表达变化。将人近端肾小管上皮细胞系 HK-2分为对照组、 IL-27组、顺铂组及顺铂 +IL-27组，实时荧光定量 PCR分析顺铂处理 0.5、1、6、12和 24 h后 IL-27受体 GP130 和 WSX-1在 mRNA水平的表达； CCK-8检测细胞活性； Hoechst染色观察细胞核形态； Western blotting检测凋亡相关蛋白 Bax、Bcl-2和多聚 ADP核糖聚合酶剪切体（ cleaved poly ADP-ribose polymerase，cleaved PARP）的表达。结果 ·顺铂组小鼠血尿素氮、血清肌酐的水平呈时间依赖性升高，肾脏组织中 p28和 Ebi3 mRNA的表达水平增加并在顺铂注射 48 h时出现峰值。顺铂处理 1 h内 HK-2细胞 GP130 和 WSX-1的表达增高，处理 6、12、24 h后 GP130和 WSX-1的表达无明显变化。与顺铂组比较，顺铂 +IL-27组细胞活力显著提升，细胞核浓缩、破裂减少，Bcl-2表达增加，Bax及 cleaved PARP表达下降。结论 · IL-27在顺铂所致 AKI中高表达且可能通过抑制细胞凋亡发挥保护作用。
关键词: 急性肾损伤, 顺铂, 白细胞介素 -27, 凋亡
Abstract:
Objective · To investigate the of interleukin-27 (IL-27) in cisplatin-induced acute kidney injury (AKI) and explore its inhibitory effect on apoptosis in AKI. Methods · C57BL/6 mice were randomly divided to control group and cisplatin group, in which mice were sacrificed at 24, 48 and 72 h after cisplatin administration. Blood urea nitrogen (BUN) and serum creatinine (Scr) levels were estimated. And the levels of mRNA for the IL-27 subunits, i.e. p28 and EBI3 in kidneys were determinedreal-time PCR. As for in vitro experiments, after cisplatin incubation for 0.5, 1, 6, 12 and 24 h, HK-2 cells, a line of proximal tubular epithelial cells, were collected to assess the of IL-27 receptors (GP130 and WSX-1) at mRNA level. After that, HK-2 cells were treated with phosphate buffer saline or recombinant human IL-27 protein after cisplatin treatment, cell viability was detectedCCK-8, nuclear morphology was observedHoechst staining, and apoptosis related proteins including Bax, Bcl-2, and cleaved poly ADP-ribose polymerase (PARP) were estimatedWestern blotting. Results · Compared with control group, BUN and Scr in cisplatin group significantly increased in a time-dependent manner after cisplatin injection. The mRNA levels of p28 and Ebi3 grew in injured kidneys, and their highest s were exhibited at 48 h after cisplatin injection. In HK-2 cells, GP130 and WSX-1 mRNA significantly increased at 1 h after cisplatin treatment but reduced to the basal level at 6, 12 and 24 h. IL-27 treatment significantly up-regulated cell viability and alleviated apoptosis and necrosis in cisplatin-treated HK-2 cells. In addition, IL-27 treatment inhibited the cleaved PARP and pro-apoptotic protein Bax, and upregulated antiapoptotic protein Bcl-2 in cisplatin-treated HK-2 cells. Conclusion · The of IL-27 increases in cisplatin-induced AKI, and it may protect against AKIreducing cell apoptosis. [Key words]acute kidney injury; cisplatin; interleukin-27; apoptosis
Key words: acute kidney injury, cisplatin, interleukin-27, apoptosis
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