CAJ | 학술논문

目的：肾素-血管紧张素系统(rennin-angiotensin system,RAS)参与糖代谢。血管紧张素转换酶2( angiotensin converting enzyme 2,ACE2)-血管紧张素(1-7)﹝Ang-(1-7)﹞-Ang-(1-7)受体(MAS)是新发现的 RAS 调节轴,可拮抗经典轴的生物效应。本文研究 ACE2/ Ang-(1-7)在肝脏糖代谢中的作用。方法①利用 Ang-(1-7)及 MAS 拮抗剂(A779)干预 HepG2细胞,或 ACE2过表达载体转染人肝细胞株 HepG2；②实时荧光定量 PCR(real-time PCR,RT-PCR )检测糖代谢相关基因和氧化应激相关基因表达；③流式细胞仪检测活性氧( reactive oxygen species,ROS)浓度；④蛋白质印迹( Western blotting)法检测还原型辅酶 II (NADPH)相关亚基的表达。结果① ACE2过表达细胞中,糖异生相关基因磷酸烯醇式丙酮酸羧激酶(phosphoenolpyruvate carboxykinase,PEPCK)的表达水平显著降低,而葡萄糖-6-磷酸激酶(glucose-6-phosphatase,G6Pase)与对照组相比差异无统计学意义；②葡萄糖转运蛋白2(glucose transporter type 2,Glut2)、胰岛素受体底物2(insulin receptor substrate,IRS-2)和腺苷酸活化蛋白激酶α亚基2﹝adenosine 5'-monophosphate(AMP)-activated protein kinase,AMPKα2﹞的表达水平显著增高,而细胞因子信号抑制因子-3(suppressor of cytokine signaling-3,SOCS-3)与对照组相比,表达量显著降低；③ Ang-(1-7)降低 HepG2细胞内和细胞外ROS 的浓度；④ Ang-(1-7)降低 NADPH 亚基 p67和 p22的表达；ACE2过表达,也能显著降低 HepG2细胞中 p22和 p47的表达,同时,ACE2对氧化应激的保护作用能被 A779抑制。结论 ACE2改善肝脏糖代谢,其机制可能与 ACE2/ Ang-(1-7)对肝脏氧化应激的保护作用有关。
목적：신소-혈관긴장소계통(rennin-angiotensin system,RAS)삼여당대사。혈관긴장소전환매2( angiotensin converting enzyme 2,ACE2)-혈관긴장소(1-7)﹝Ang-(1-7)﹞-Ang-(1-7)수체(MAS)시신발현적 RAS 조절축,가길항경전축적생물효응。본문연구 ACE2/ Ang-(1-7)재간장당대사중적작용。방법①이용 Ang-(1-7)급 MAS 길항제(A779)간예 HepG2세포,혹 ACE2과표체재체전염인간세포주 HepG2；②실시형광정량 PCR(real-time PCR,RT-PCR )검측당대사상관기인화양화응격상관기인표체；③류식세포의검측활성양( reactive oxygen species,ROS)농도；④단백질인적( Western blotting)법검측환원형보매 II (NADPH)상관아기적표체。결과① ACE2과표체세포중,당이생상관기인린산희순식병동산최격매(phosphoenolpyruvate carboxykinase,PEPCK)적표체수평현저강저,이포도당-6-린산격매(glucose-6-phosphatase,G6Pase)여대조조상비차이무통계학의의；②포도당전운단백2(glucose transporter type 2,Glut2)、이도소수체저물2(insulin receptor substrate,IRS-2)화선감산활화단백격매α아기2﹝adenosine 5'-monophosphate(AMP)-activated protein kinase,AMPKα2﹞적표체수평현저증고,이세포인자신호억제인자-3(suppressor of cytokine signaling-3,SOCS-3)여대조조상비,표체량현저강저；③ Ang-(1-7)강저 HepG2세포내화세포외ROS 적농도；④ Ang-(1-7)강저 NADPH 아기 p67화 p22적표체；ACE2과표체,야능현저강저 HepG2세포중 p22화 p47적표체,동시,ACE2대양화응격적보호작용능피 A779억제。결론 ACE2개선간장당대사,기궤제가능여 ACE2/ Ang-(1-7)대간장양화응격적보호작용유관。
Objective Renin angiotensin system(RAS) is involved in glucose metabolism. This study evaluated the effect of the new pathway of RAS, ACE2 / Ang-(1-7), on glucose metabolism in hepatic cells. Methods ① HepG2 cells were treated with Ang-(1-7) or A779, or transfected with ACE2 over expression plasmid DNA; ② The expression of PEPCK, G6Pase, Glut2, IRS-2, AMPKα2, SOCS-3, p22 and p67 were detected by RT-PCR; ③ ROS was measured by DHE; ④ Western blotting was used to study p22 and p67. Results① In ACE2-overexpressing HepG2 cells, the expression of PEPCK was decreased; ② SOCS-3 was decreased, while Glut2, IRS-2 and AMPKα2 were increased significantly; ③ Ang-(1-7) reduces ROS production; ④ Ang-(1-7) depressed the expression of p22 and p67 in HepG2 cells, and ACE2 inhibited relative protein expression levels of p47phox and p22phox . Conclusion ACE2 / Ang-(1-7) could be involved in improving glucose metabolism via anti-oxidative effects.