虾青素通过调控线粒体自噬改善HK-2细胞纤维化  

Astaxanthin improves HK-2 cell fibrosis by regulating mitochondrial autophagy

作　　者：洪梦琪 李新科 赵宇 聂振禹 陈争跃 HONG Mengqi;LI Xinke;ZHAO Yu;NIE Zhenyu;CHEN Zhengyue(Ningbo Ninth Hospital,Ningbo 315020,Zhejiang,China)

机构地区：[1]宁波市第九医院,宁波315020 [2]宁波市鄞州区第二医院

出　　处：《现代实用医学》2024年第12期1548-1552,F0003,共6页Modern Practical Medicine

基　　金：宁波市卫生健康科技计划项目(2023Y40);浙江省医药卫生科技计划项目(2024KY1607);宁波市科技计划项目(2021J024)。

摘　　要：目的探讨虾青素(AST)改善转化生长因子-β1(TGF-β1)诱导的人肾小管上皮细胞(HK-2)纤维化的作用机制。方法将HK-2细胞随机分为正常对照组(Control组)、TGF-β1组、TGF-β1+20μg/ml AST组及TGF-β1+40μg/ml AST组,CCK-8法检测细胞活力,线粒体膜电位检测试剂盒检测线粒体膜电位水平,实时荧光定量PCR测定细胞α平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)mRNA的表达水平,Westernblot检测α-SMA、FN、微管相关蛋白轻链3(LC3)-11/1、P62、丝氨酸/苏氨酸蛋白激酶(PINK1)及帕金蛋白(parkin)蛋白的表达水平。结果与Control组相比,TGF-β1组α-SMA、FN的mRNA及蛋白水平增高,细胞活力及线粒体膜电位显著下降;在加入20、40μg/ml的AST后,α-SMA、FN的mRNA及蛋白水平下降,细胞活力及线粒体膜电位显著升高(均P<0.05)。此外,与Control组相比,TGF-β1组LC3-Ⅱ/Ⅰ、parkin、PINK1蛋白水平显著升高,P62蛋白水平显著降低(均P<0.05);与TGF-β1组相比,TGF-β1+20μg/ml AST组及TGF-β1+40μg/ml AST组LC3-II/1、parkin、PINK1蛋白水平显著降低,P62蛋白水平显著升高(均P<0.05)。结论AST可能通过调节线粒体自噬来改善TGF-β1诱导的HK-2细胞纤维化。Objective To investigate the mechanism of astaxanthin(AST)in improving transforming growth factorβ1(TGF-β1)induced fibrosis in human renal tubular epithelial cells(HK-2).Methods HK-2 cells were divided into control group,TGF-β1 group,TGF-β1+20μg/ml AST group,and TGF-β1+40μg/ml AST group.Cell viability was assessed by CCK-8,and mitochondrial membrane potential level was detected by mitochondrial membrane potential detection kit.The mRNA expressions ofα-smooth muscle actin(α-SMA),fibronectin(FN)were detceted by quantitative real time polymerase chain reaction(qRT-PCR).The protein expressions ofα-SMA,FN,microtubule-associated protein light chain 3(LC3)-Ⅱ/Ⅰ,P62,PINK1,and parkin were detected by Western blot.Results The mRNA,protein expressions ofα-SMA,FN in TGF-β1 group were higher than those of control group;and cell viability,mitochondrial membrane potential level were lower than those of control group(all P<0.05).After treatment with 20μg/ml or 40μg/ml AST in the TGF-β1 group;the mRNA,protein expressions ofα-SMA,FN were significantly decreased;while cell viability,mitochondrial membrane potential level were significantly increased(all P<0.05).The protein expressions of LC3-Ⅱ/Ⅰ,parkinl,and PINK1 were higher than those of control group,and the protein expression of P62 was lower than that of control group(all P<0.05).Compared to the TGF-β1 group,the protein expressions of LC3-Ⅱ/Ⅰ,parkinl,and PINK1 were significantly decreased,and the protein expression of P62 was significantly increased in the TGF-β1+20μg/ml AST group and TGF-β1+40μg/ml AST group(all P<0.05).Conclusions AST can alleviate TGF-β1-induced fibrosis in HK-2 cells by regulating mitophagy.

关 键 词：虾青素 肾小管上皮细胞 线粒体自噬 纤维化 

分 类 号：R692[医药卫生—泌尿科学]