Roxadustat对肾缺血再灌注损伤的保护作用及机制研究

目的:探讨一种新型PHD抑制剂Roxadustat对小鼠肾缺血再灌注损伤的保护作用及其可能的作用机制。方法:将雄性C57BL/6小鼠随机分为4组:假手术组(sham)、损伤组(IR)、损伤+低剂量给药组(IR+Rox10 mg/kg)以及损伤+高剂量组(IR+Rox25 mg/kg)。除假手术组外,其余各组分别于造模前1h、6h、12h给药,并于造模后6h、12h、24h、48h采血检测血肌酐(Scr)、尿素氮(BUN),1d、2d、5d取材肾脏进行病理检测。此外,利用HK-2细胞建立缺氧模型,测定给药后细胞活力和细胞凋亡情况的变化及凋亡通路蛋白和HIF-1α的表达情况。结果:与sham组和IR组相比,给药组Scr和BUN水平均明显降低,且高剂量组Scr和BUN水平显著低于低剂量组,且给药组形态学损伤更轻,细胞凋亡明显减少。细胞学实验显示,Roxadustat能提高低氧条件下HK-2细胞的活力,降低细胞凋亡,并抑制低氧导致的Bax升高,提高Bcl-2的表达,而用HIF-1α抑制剂2-MeOE2,可消除Roxadustat对凋亡的抑制作用。结论:Roxadustat能够通过上调HIF-1α表达,抑制线粒体途径凋亡通路相关蛋白表达,减少细胞凋亡,对小鼠肾脏缺血再灌注损伤产生保护作用。

Protective Effect and Mechanism of Roxadustat on the Renal Ischemia-reperfusion Injury

Objective:To investigate the effect of a new PHD inhibitor Roxadustat on renal ischemia-reperfusion injury in mice and its possible mechanism.Methods:Male C57 BL/6 mice were randomly divided into four groups:sham operation group(sham),injury group(IR),injury+low dose group(IR+Rox10 mg/kg)and injury+high dose group(IR+Rox25 mg/kg).Except for the sham operation group,the other groups were given drugs at 1 h,6 h and 12 h before modeling,and blood samples were taken at 6 h,12 h,24 h and 48 h after modeling to detect serum creatinine(Scr)and urea nitrogen(BUN),1 d,2 d and 5 d for pathological examination.In addition,the hypoxia model was established by HK-2 cells,and the changes of cell viability and apoptosis,as well as the expression of apoptosis pathway protein and HIF-1αwere measured.Result(s):Compared with sham group and IR group,the levels of Scr and BUN in high dose group were significantly lower than those in low dose group,and the levels of Scr and BUN in high dose group were significantly lower than those in low dose group,and the morphological damage was lighter and the apoptosis was significantly reduced in the treatment group.Cytological experiments showed that Roxadustat could increase the vitality of HK-2 cells under hypoxia,reduce apoptosis,inhibit the increase of Bax induced by hypoxia and increase the expression of Bcl-2,while HIF-1αinhibitor 2 MeOE2,It can eliminate the inhibitory effect of Roxadustat on apoptosis.Conclusions:Roxadustat can protect the kidney from ischemia-reperfusion injury in mice through up-regulating the expression of HIF-1α,inhibiting the expression of apoptosis pathway-related proteins and reducing apoptosis.