CAJ | 학술논문

目的：探讨槲皮素对血管紧张素Ⅱ（angiotensinⅡ，AngⅡ）诱导的心肌细胞肥大的防治作用及机制。方法：分别在原代新生大鼠心肌细胞和培养的H9c2心肌细胞，用100 nmol／L的AngⅡ诱导心肌细胞肥大模型，并给予3种不同浓度的槲皮素（10μmol／L，20μmol／L，40μmol／L）处理，利用免疫荧光检测原代和H9c2心肌细胞表面积的改变，采用荧光底物法检测H9c2心肌细胞蛋白酶体的活性变化，以及用Western blot检测H9c2心肌细胞糖原合酶激酶（ GSK）－3α／β、Akt及其磷酸化情况。结果：与对照组相比，模型组原代心肌细胞和H9c2心肌细胞表面积均显著增大，槲皮素处理组2种心肌细胞表面积较模型组均明显减小，而20μmol／L槲皮素组减小更明显（P＜0．05）。在H9c2细胞实验中发现模型组蛋白酶体的糜蛋白酶样、半胱天冬酶样和胰蛋白酶样活性明显升高，20μmol／L和40μmol／L槲皮素组半胱天冬酶样和胰蛋白酶样活性较模型组均明显降低，而糜蛋白酶样活性只有在槲皮素20μmol／L时有显著差异（ P＜0．05）；Western blot 检测结果显示，模型组磷酸化（ p）－GSK－3α、p－GSK－3β和p－Akt水平较对照组均明显增加，而20μmol／L和40μmol／L槲皮素处理均使p－GSK－3α、p－GSK－3β和p－Akt水平明显下降（P＜0．05）。结论：槲皮素可明显抑制蛋白酶体活性从而减轻心肌细胞肥大，其机制可能是通过下调Akt活性而升高GSK－3α／β活性实现的。
목적：탐토곡피소대혈관긴장소Ⅱ（angiotensinⅡ，AngⅡ）유도적심기세포비대적방치작용급궤제。방법：분별재원대신생대서심기세포화배양적H9c2심기세포，용100 nmol／L적AngⅡ유도심기세포비대모형，병급여3충불동농도적곡피소（10μmol／L，20μmol／L，40μmol／L）처리，이용면역형광검측원대화H9c2심기세포표면적적개변，채용형광저물법검측H9c2심기세포단백매체적활성변화，이급용Western blot검측H9c2심기세포당원합매격매（ GSK）－3α／β、Akt급기린산화정황。결과：여대조조상비，모형조원대심기세포화H9c2심기세포표면적균현저증대，곡피소처리조2충심기세포표면적교모형조균명현감소，이20μmol／L곡피소조감소경명현（P＜0．05）。재H9c2세포실험중발현모형조단백매체적미단백매양、반광천동매양화이단백매양활성명현승고，20μmol／L화40μmol／L곡피소조반광천동매양화이단백매양활성교모형조균명현강저，이미단백매양활성지유재곡피소20μmol／L시유현저차이（ P＜0．05）；Western blot 검측결과현시，모형조린산화（ p）－GSK－3α、p－GSK－3β화p－Akt수평교대조조균명현증가，이20μmol／L화40μmol／L곡피소처리균사p－GSK－3α、p－GSK－3β화p－Akt수평명현하강（P＜0．05）。결론：곡피소가명현억제단백매체활성종이감경심기세포비대，기궤제가능시통과하조Akt활성이승고GSK－3α／β활성실현적。
AIM:ToinvestigatetheprotectiveeffectofquercetinonangiotensinⅡ(AngⅡ)-inducedcardio-myocyte hypertrophy and its possible mechanism .METHODS: Cardiomyocyte hypertrophy was induced by AngⅡ ( 100 nmol/L) in primary neonatal cardiomyocytes and H 9c2 cells.The cells were treated with different concentration of querce-tin (10 μmol/L, 20 μmol/L and 40 μmol/L) for 48 h and then the cardiomyocyte surface areas were measured by immu-nofluorescence .Proteasome activity was detected by fluorescent peptide substrate .The phosphorylated levels of GSK-3α/βand Akt in H9c2 cells were determined by Western blot .RESULTS:Compared with control group , the cardiomyocyte sur-face areas were both increased in primary cultured neonatal cardiomyocytes and H 9c2 cells, while the surface areas were significantly decreased by quercetin , especially at concentration of 20 μmol/L compared with Ang Ⅱgroup (P<0.05). Compared with control group , the chymotrypsin-like, trypsin-like and caspase-like activities of proteasome were all in-creased in H9c2 cells (P<0.05).The trypsin-like and caspase-like activities of proteasome were inhibited by 20 μmol/L and 40 μmol/L quercetin , while chymotrypsin-like activity was inhibited only at 20 μmol/L of quercetin compared with AngⅡgroup (P<0.05).In addition, phosphorylated levels of GSK-3α-Ser21, GSK-3β-Ser9 and Akt-Ser473 in AngⅡgroup were all increased compared with control group , which were obviously inhibited by in 20 μmol/L and 40 μmol/L quercetin ( P<0.05 ) .CONCLUSION: Quercetin decreases cardiomyocyte hypertrophy through proteasome inhibition , which may be related to the inhibition of Akt and therefore increasing activation of GSK -3α/βin H9c2 cells.