CAJ | 학술논문

目的探讨人脂联素重组慢病毒( Lenti-apM1 -EGFP)对血小板源性生长因子(PDGF)诱导的肾小球系膜细胞(HMC)增殖及腺苷酸活化蛋白激酶(AMPK)信号通路活化的影响.方法将Lenti-apM1-EGFP感染HMC,确定最佳感染复数(MOI),并观察其细胞毒性.胸腺嘧啶核苷(3 H-TdR)掺入法检测PDGF-BB及AMPK抑制剂Compound C处理过的HMC及Lenti-apM1-EGFP感染后HMC增殖,流式细胞仪检测细胞周期,Western印迹检测AMPK磷酸化水平.结果 Lenti-apM 1-0EGFP对HMC无明显毒性；MOI为50的Lenti-apM1-EGFP能够有效地感染HMC,并使之稳定表达脂联素蛋白(149.6±12.8).μg/L.PDGFBB以剂量依赖性的方式促进HMC增殖,影响细胞周期；对Lenti-apM1 -EGFP感染后HMC则没有此效应,同时应用Compound C后HMC增殖能力增强.Lenti-apM1 -EGFP感染后HMC的AMPK磷酸化水平显著升高.结论脂联素通过激活AMPK信号通路抑制PDGF-BB诱导的系膜细胞增殖.
목적 탐토인지련소중조만병독( Lenti-apM1 -EGFP)대혈소판원성생장인자(PDGF)유도적신소구계막세포(HMC)증식급선감산활화단백격매(AMPK)신호통로활화적영향.방법 장Lenti-apM1-EGFP감염HMC,학정최가감염복수(MOI),병관찰기세포독성.흉선밀정핵감(3 H-TdR)참입법검측PDGF-BB급AMPK억제제Compound C처리과적HMC급Lenti-apM1-EGFP감염후HMC증식,류식세포의검측세포주기,Western인적검측AMPK린산화수평.결과 Lenti-apM 1-0EGFP대HMC무명현독성；MOI위50적Lenti-apM1-EGFP능구유효지감염HMC,병사지은정표체지련소단백(149.6±12.8).μg/L.PDGFBB이제량의뢰성적방식촉진HMC증식,영향세포주기；대Lenti-apM1 -EGFP감염후HMC칙몰유차효응,동시응용Compound C후HMC증식능력증강.Lenti-apM1 -EGFP감염후HMC적AMPK린산화수평현저승고.결론 지련소통과격활AMPK신호통로억제PDGF-BB유도적계막세포증식.
Objective To evaluate the effects of recombinant lentivirus encoding human apM1 gene ( LentiapM1-EGFP) on platelet-derived growth factor (PDGF) induced human mesangial cell (HMC) proliferation and intracellular AMP-activated protein kinase(AMPK) signaling pathway.Methods Protein expression of apM1 in cell culture supernatant of HMC transfected with Lenti-apM1-EGFP was detected by ELISA.The effect of human adiponectin on cell proliferation and cell cycle was assessed by [ 3 H ] thymidine incorporation assay and Flow cytometry respectively.The phosphorylation of AMPK was detected by Western blotting.Results Lenti-apM1-EGFP had no significant toxicity on HMC.The 50 multiplicity of infection (MOI) of the Lenti-apM1-EGFP efficiently infected HMC,and made it stable expression of adiponectin protein ( 149.6 ± 12.8 ) μg/L.PDGF-induced HMCs proliferation was significantly inhibited by adiponectin.When co-treatment with compound C,an AMPK inhibitor,the inhibitory effort was reversed.The phosphorylation level of AMPK was increased in HMC transfected Lenti-apM1-EGFP compared to that of control.Conclusions Adiponectin antagonizes stimulatory effect of platelet-derived growth factor on mesangial cell proliferation by AMPK signaling.