吴茱萸碱对高糖诱导的滋养层细胞增殖、迁移、侵袭的影响及机制

Impact of evodiamine on the proliferation,migration and invasion of trophoblastic cells induced by high glucose and its mechanism

目的 基于晚期糖基化终末产物(AGE)/晚期糖基化终末产物受体(RAGE)信号通路探讨吴茱萸碱对高糖诱导的滋养层细胞增殖、迁移、侵袭能力的影响及潜在机制。方法 将人滋养层细胞HTR-8/SVneo分为对照组、高糖组、吴茱萸碱低浓度组(2μmol/L)、吴茱萸碱高浓度组(4μmol/L)、pc-NC组(转染pc-NC质粒+4μmol/L吴茱萸碱)、pc-RAGE组(转染pc-RAGE质粒+4μmol/L吴茱萸碱)。除对照组外的其余各组细胞均在高糖(25 mmol/L葡萄糖)环境下培养,除对照组、高糖组外的其余各组细胞均转染相应质粒和(或)接受相应药液干预。检测各组细胞的存活率、凋亡率、划痕愈合率、侵袭数,以及AGE、RAGE、核因子κB p65(NF-κB p65)、基质金属蛋白酶2(MMP-2)、MMP-9蛋白及mRNA的表达情况。结果 与对照组比较,高糖组细胞的存活率、划痕愈合率、侵袭数和MMP-2、MMP-9蛋白及m RNA的表达水平均显著降低(P<0.05),其凋亡率,AGE、RAGE蛋白及mRNA的表达水平,NF-κB p65 mRNA的表达水平和NF-κB p65蛋白的磷酸化水平均显著升高(P<0.05);与高糖组比较,吴茱萸碱低、高浓度组细胞上述指标均显著改善,且高浓度组的效果显著优于低浓度组(P<0.05);过表达RAGE会减弱吴茱萸碱对高糖诱导的滋养层细胞上述指标(AGE mRNA及蛋白除外)的改善作用(P<0.05)。结论 吴茱萸碱可促进高糖诱导的滋养层细胞增殖、迁移、侵袭,改善其功能损伤,上述作用与抑制AGE/RAGE信号通路有关。

OBJECTIVE To investigate the impact of evodiamine on the proliferation,migration and invasion abilities of trophoblastic cells induced by high glucose and its potential mechanism based on advanced glycation end products(AGE)/receptor for advanced glycation end products(RAGE) signaling pathway.METHODS Human trophoblastic cells HTR-8/SVneo were divided into control group,high glucose group,evodiamine low-dose group(2 μmol/L),evodiamine high-dose group(4 μmol/L),pc-NC group(transfected with pc-NC plasmid+4 μmol/L evodiamine),and pc-RAGE group(transfected with pc-RAGE plasmid+4 μmol/L evodiamine).Cells in all groups except for the control group were cultured in a high sugar(25 mmol/L glucose) environment,and cells in all groups except for the control group and the model group were transfected with the corresponding plasmids and/or received the corresponding drug interventions.The survival rate,apoptotic rate,scratch healing rate,and invasion number,as well as the protein and mRNA expressions of AGE,RAGE,nuclear factor-κB p65(NF-κB p65),matrix metalloproteinase-2(MMP-2),and MMP-9 were examined in each group.RESULTS Compared with the control group,the cell survival rate,scratch healing rate,invasions number,and the mRNA and protein expressions of MMP-2 and MMP-9 in the high glucose group significantly decreased(P<0.05),while the apoptotic rate,the mRNA and protein expressions of AGE and RAGE,the mRNA expression of NF-κB p65,and the phosphorylation level of NF-κB p65 protein significantly increased(P<0.05);compared with the high glucose group,the above indexes of cells in evodiamine low-dose and high-dose groups were significantly improved,and the effect of the high-dose group was significantly better than that of the low-dose group(P<0.05);overexpression of RAGE attenuated the ameliorative effect of evodiamine on the above indexes in high glucose-induced trophoblast cells(except for AGE m RNA and protein)(P<0.05).CONCLUSIONS Evodiamine can promote the proliferation,migration and invasion of high glucose-induced trophoblast cells and ameliorate their functional impairment,and the above effects are associated with the inhibition of the AGE/RAGE signaling pathway.