褪黑素对PDGF-BB激活的HSCs自噬作用的研究

Effect of melatonin on autophagy in PDGF-BB induced activated hepatic stellate cells

目的探究褪黑素对血小板源性生长因子BB(PDGF-BB)激活的肝星状细胞(HSCs)自噬作用的影响。方法HSCs(HSC-T6)被分为5组:对照组、模型组和实验组(分为3组:褪黑素1 nmol/L、1μmol/L和0. 1 mmol/L组),培养24 h贴壁后以无血清培养基培养,模型组及不同实验组加入血小板源性生长因子(PDGF-BB),实验组加入不同浓度的褪黑素,培养48 h。MTT检测褪黑素对HSCs增殖的影响; Western blot检测PDGF及褪黑素对HSCs自噬的作用。结果相对于实验组,褪黑素能抑制HSCs的增殖(P <0. 05)。PDGF-BB能明显上调HSCsα平滑肌肌动蛋白(α-SMA)、Beclin1、微管相关蛋白1轻链3Ⅱ型(LC3Ⅱ)及血小板衍生生长因子受体(PDGFR)的表达,与模型组相比,褪黑素能抑制HSCsα-SMA、Beclin1、LC3Ⅱ及PDGFR的表达(P<0. 05)。结论褪黑素可抑制HSCs的活化,可能通过抑制HSCs内自噬表达发挥作用。

Objective To investigate the effect of melatonin on autophagy induced by platelet derived growth factor-BB(PDGF-BB)in hepatic stellate cells.Methods The HSC-T6 cells were divided into five groups:control group,model group and experimental groups(three groups,concentration of melatonin of 1 nmol/L,1μmol/L and 0.1 mmol/L).After being cultured for 24 h,they were replaced with FBS-free medium and treated with PDGF-BB(10 ng/ml)excepted the control group,and different concentrations(1 nmol/L,1μmol/L and 0.1 mmol/L)were added immediately in three expremental groups.After incubation for 48 h,MTT assay was performed to assess the proliferation of hepatic stellate cells,Western blot were performed to assess the levels of autophagy.Results Comparing with the model group,melatonin could inhibit the proliferation of hepatic stellate cells(P<0.05).Additionally,in PDGF-BB treated group,the levels ofα-smooth muscle actin(α-SMA),Beclin1,microtubule-associated protein 1 light chain-3Ⅱ(LC3Ⅱ),platelet derived growth factor receptor(PDGFR)were up-regulated,which were down-regulated after administration of melatonin(P<0.05).Conclusion Melatonin can alleviate the activation of hepatic stellate cells,which may relate to the level of autophagy.