摘要
目的:探讨虾青素(astaxanthin,AST)减轻大鼠对比剂急性肾损伤(contrast-induced acute kidney injury,CI-AKI)的作用机制。方法:雄性SD大鼠随机分为5组:假手术(sham)组、造模组(CI-AKI组)、AST治疗组(AST+CI-AKI组)、自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)干预组(3-MA+CI-AKI组)和AST联合自噬抑制剂组(AST+3-MA+CI-AKI组),每组10只。建立大鼠CI-AKI模型后72 h收集血清和肾脏组织,检测大鼠血清肌酐(serum creatinine,SCr)和血尿素氮(blood urea nitrogen,BUN)水平,检测肾脏组织总超氧化物歧化酶(total superoxide dismutase,T-SOD)和丙二醛(malondialdehyde,MDA)水平;HE染色观察肾脏结构;活性氧簇(reactive oxygen species,ROS)染色观察肾组织冰冻切片ROS水平;Western blot检测线粒体自噬及细胞自噬相关蛋白PTEN诱导假定激酶1(PTEN-induced putative kinase 1,PINK1)、parkin、微管相关蛋白1轻链3(microtubule-associated protein 1light chain 3,LC3)和溶酶体相关膜蛋白2(lysosomal-associated membrane protein 2,LAMP2)的表达水平;免疫荧光双染检测LC3标记的自噬小体与LAMP2标记的溶酶体或热休克蛋白60(heat shock protein 60,Hsp60)标记的线粒体的共定位情况,检测线粒体自噬及细胞自噬活性;透射电镜观察自噬小体及线粒体形态。结果:与sham组相比,CI-AKI组SCr、BUN、MDA和ROS水平及PINK1、parkin、LC3 II/LC3 I和LAMP2表达水平均显著升高,T-SOD水平显著降低(P<0.05);与CI-AKI组相比,AST+CI-AKI组SCr、BUN、MDA和ROS水平均显著降低(P<0.05),T-SOD水平及PINK1、parkin、LC3 II/LC3 I和LAMP2表达水平均显著升高(P<0.05),3-MA+CI-AKI组SCr、BUN、MDA和ROS水平显著升高(P<0.05),T-SOD水平及PINK1、parkin、LC3 II/LC3 I和LAMP2表达水平均显著下降(P<0.05)。结论:AST通过上调PINK1/parkin通路增强线粒体自噬水平,减少ROS释放,从而减轻大鼠CI-AKI。
AIM:To investigate the effect of astaxanthin(AST)on contrast-induced acute kidney injury(CI-AKI)in rats.METHODS:Male SD rats were randomly divided into 5 groups:sham group,CI-AKI group,AST treat-ment(AST+CI-AKI)group,autophagy inhibitor 3-methyladenine(3-MA)intervention(3-MA+CI-AKI)group,and AST combined with autophagy inhibitor(AST+3-MA+CI-AKI)group,with 10 rats in each group.The rats were sacrificed 72 h after modeling,and the serum and kidney tissues were collected.The serum creatinine(SCr)and blood urea nitrogen(BUN)levels were measured.Total superoxide dismutase(T-SOD)and malondialdehyde(MDA)levels in kidney tissues were measured.HE staining was used to observe the structure of kidney.Frozen section was used to observe the generation of reaction oxygen species(ROS)in renal tissues by ROS staining.The expression of mitophagy-and cellular autophagy related proteins PTEN-induced putative kinase 1(PINK1),parkin,microtubule-associated protein 1 light chain 3(LC3)and lysosomal-associated membrane protein 2(LAMP2)was determined by Western blot.The co-location of LC3-labeled autophagosomes with heat shock protein 60(Hsp60)-labeled mitochondria or LAMP2-labeled lysosomes was observed by immunofluorescence double staining.Transmission electron microscopy was used to observe the morphological changes of autophagosomes and mitochondria.RESULTS:Compared with sham group,the levels of SCr,BUN,MDA and ROS,and the expression of PINK1,parkin,LC3 II/LC3 I and LAMP2 were increased in CI-AKI group(P<0.05),while the level of T-SOD was decreased(P<0.05).Compared with CI-AKI group,the levels of SCr,BUN,MDA and ROS were decreased in AST+CI-AKI group(P<0.05),while the level of T-SOD,and the expression of PINK1,parkin,LC3 II/LC3 I and LAMP2 were increased(P<0.05).Compared with CI-AKI group,the levels of SCr,BUN,MDA and ROS were increased in 3-MA+CI-AKI group(P<0.05),while the level of T-SOD,and the expression of PINK1,parkin,LC3 II/LC3 I and LAMP2 were decreased(P<0.05).CONCLUSION:Astaxanthin attenuates CI-AKI in rats via PINK1/parkin
作者
张驰昊
郑迪
张成
轩永丽
李文华
ZHANG Chi-hao;ZHENG Di;ZHANG Cheng;XUAN Yong-li;LI Wen-hua(Institute of Cardiovascular Diseases,Xuzhou Medical University,Xuzhou 221000,China;Department of Cardiology,Af-filiated Hospital of Xuzhou Medical University,Xuzhou 221000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2022年第8期1430-1439,共10页
Chinese Journal of Pathophysiology
基金
江苏省研究生科研与实践创新计划项目(No.KYCX21_2672)
徐州医科大学附属医院院级科研项目(No.2020KB004)。
