人脂联素基因重组慢病毒对血小板源性生长因子诱导的肾小球系膜细胞增殖的影响

Effects of recombinant lentivirus encoding human apM1 gene on platelet-derived growth factor induced mesangial cell proliferation

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（。H—TdR）掺入法检测PDGF—BB及AMPK抑制剂CompoundC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-apM1-OEGFP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。PDGF．BB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CompoundC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。

Objective To evaluate the effects of recombinant lentivirus encoding human apM1 gene （ Lenti- apM1-EGFP） on platelet-defived 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 ） ~g/L. PDGF-induced HMCs proliferation was significantly inhibited by adiponeetin. 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.