CAJ | 학술논문

目的：探讨异鼠李素对氧化应激诱导的心肌细胞损伤的保护作用及其作用机制。方法用MTT实验确定异鼠李素的心肌细胞毒作用浓度,以及抗H2 O2氧化应激损伤的最佳作用浓度。将实验组分为正常组、模型组、治疗组、单给药组。治疗组、单给药组均用最佳浓度的异鼠李素预孵育12 h,模型组、治疗组在预孵育结束后用300μmol · L-1 H2 O2作用4 h,模拟心肌细胞的氧化应激损伤。利用流式细胞术检测心肌细胞凋亡细胞比例、ROS产生等指标,利用荧光显微镜检测细胞的线粒体膜电位去极化,利用荧光酶标仪检测抗氧化酶与氧化产物MDA产量,利用Western blot检测线粒体凋亡信号通路与 Nrf2/ARE 信号通路的相关蛋白表达。结果与正常组相比,单给药组心肌细胞的凋亡细胞比例, ROS产生,线粒体膜电位去极化,细胞质细胞色素 C、caspase-9、caspase-3、Bcl-2、Bax 表达,抗氧化酶与氧化产物MDA产量差异均无显著性( P>0．05)；与模型组相比,治疗组心肌细胞凋亡比例明显降低(P<0．01),ROS产生明显降低(P<0．05),线粒体膜电位去极化明显改善(P<0．01),细胞质细胞色素C、caspase-9、caspase-3、Bax表达明显降低( P<0．01),Bcl-2表达增加(P<0．01),抗氧化酶表达增加(P<0．01),氧化产物MDA表达降低(P<0．01)；心肌细胞中Nrf2核转位与HO-1表达均随着异鼠李素预孵育时间的延长而增加。结论异鼠李素能够保护H2 O2引起的心肌细胞损伤。其机制为影响线粒体凋亡通路以及激活Nrf2/ARE信号通路,实现抗氧化、抗凋亡功能。
목적：탐토이서리소대양화응격유도적심기세포손상적보호작용급기작용궤제。방법용MTT실험학정이서리소적심기세포독작용농도,이급항H2 O2양화응격손상적최가작용농도。장실험조분위정상조、모형조、치료조、단급약조。치료조、단급약조균용최가농도적이서리소예부육12 h,모형조、치료조재예부육결속후용300μmol · L-1 H2 O2작용4 h,모의심기세포적양화응격손상。이용류식세포술검측심기세포조망세포비례、ROS산생등지표,이용형광현미경검측세포적선립체막전위거겁화,이용형광매표의검측항양화매여양화산물MDA산량,이용Western blot검측선립체조망신호통로여 Nrf2/ARE 신호통로적상관단백표체。결과여정상조상비,단급약조심기세포적조망세포비례, ROS산생,선립체막전위거겁화,세포질세포색소 C、caspase-9、caspase-3、Bcl-2、Bax 표체,항양화매여양화산물MDA산량차이균무현저성( P>0．05)；여모형조상비,치료조심기세포조망비례명현강저(P<0．01),ROS산생명현강저(P<0．05),선립체막전위거겁화명현개선(P<0．01),세포질세포색소C、caspase-9、caspase-3、Bax표체명현강저( P<0．01),Bcl-2표체증가(P<0．01),항양화매표체증가(P<0．01),양화산물MDA표체강저(P<0．01)；심기세포중Nrf2핵전위여HO-1표체균수착이서리소예부육시간적연장이증가。결론이서리소능구보호H2 O2인기적심기세포손상。기궤제위영향선립체조망통로이급격활Nrf2/ARE신호통로,실현항양화、항조망공능。
Aim To investigate the protective effect of isorhamnetin on H9 C2 myocardial cell line and its mechanisms. Methods The toxicity and optimal pro-tective concentration of isorhamnetin were determined by MTT assay. The experimental subjects were divided into four groups:group N ( normal ) , group M ( mod-el) , group M + ISO ( model + isorhamnetin ) , and group ISO ( isorhamnetin only ) . Group M +ISO and ISO were pre-incubated with isorhamnetin for 12 hours while other groups with plain DMEM. Group M and M+ ISO were treated with 300μmol · L-1 H2 O2 for 4 hours after pre-incubation. Mitochondrial membrane potential depolarization of H9 C2 was measured by fluo-rescence microscope. Apoptotic rate and ROS produc-tion of injured myocardial cell line were detected using <br> flow cytometry. The oxidative indictors were measured by spectrophotometry. The expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bcl-2, Bax, Nrf2 and HO-1 were examined by Western blot. Result There was no difference in mitochondrial membrane potential depolarization, apoptotic rate, ROS produc-tion, oxidative indictors production and expressions of cytoplasmic cytochrome C, caspase-9,caspase-3, Bcl-2 , Bax between groups ISO and N ( P>0. 05 ) . Apop-totic rate, ROS production, expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bax, MDA pro-duction of group M+ISO were significantly lower than those of group M ( P < 0. 01 ) . And mitochondrial membrane potential, Bcl-2, CAT, SOD and GSH-Px of group M + ISO were increased compared to group M . <br> Nuclear translocation of Nrf2 and expression of HO-1 in myocardial cell line were increased with the prolonged isorhamnetin incubation time. Conclusion Isorham-netin could protect myocardial cell line against H2 O2-induced oxidative injury and apoptosis through the in- <br> terruption of mitochondrial dependent apoptotic path-way and activation of Nrf2/ARE pathway.