表没食子儿茶素没食子酸酯对人脐带动脉平滑肌细胞增殖的抑制作用

目的: 考察表没食子儿茶素没食子酸酯(epigallocatechingallate,EGCG)对体外培养的人脐带动脉平滑肌细胞(HUASMCs)增殖的抑制作用及作用机制,为临床治疗相关的心血管疾病提供参考。方法: 组织块贴壁法原代培养HUASMCs,选传至3代的细胞进行免疫组织化学染色法鉴定;MTT比色法,测定EGCG(109.08 μmol·L^-1)在不同作用时间(3~96 h)和不同作用浓度(1.09~1 090.75 μmol·L^-1)下对HUASMCs增殖的抑制作用;RT-PCR法,测定EGCG(2.18,21.82,218.15 μmol·L^-1)对HUASMCs表达IL-6 mRNA的影响;ELISA法,测定EGCG(2.18,21.82,218.15 μmol·L^-1)对HUASMCs分泌IL-6水平的影响。结果: EGCG在各个检测时间点均可抑制体外培养HUASMCs的增殖(P<0.01),3~48 h抑制作用随作用时间延长而增加,48 h增殖抑制最强,72 h,96 h与48 h增殖抑制作用相比较,差异无统计学意义(P>0.05)。EGCG各浓度对HUASMCs体外增殖均有抑制作用(P<0.05或P<0.01),EGCG呈剂量依赖性,IC₅₀为413.39 μmol·L^-1。EGCG(21.82,218.15 μmol·L^-1)对HUASMCs表达IL-6 mRNA有抑制作用(P<0.01)。EGCG(2.18,21.82,218.15 μmol·L^-1)可抑制HUASMCs分泌IL-6(P<0.05)。结论: EGCG对HUASMCs的体外增殖有抑制作用。EGCG可抑制HUASMCs IL-6 mRNA的表达和IL-6因子的分泌。

Inhibiting effects of Epigallocatechingallate on proliferation of human umbilical artery smooth muscle cells

OBJECTIVE To investigate the inhibiting effects and relevant mechanism of Epigallocatechingallate (EGCG) on proliferation of human umbilical artery smooth muscle cells (HUASMCs), providing reference for clinical treatment against cardiovascular diseases. METHODS HUASMCs were primarily cultured by using tissue adherent method and the third generation of cells were selected for immunohistochemical staining. The inhibiting effects of EGCG (109.08 μmol·L^-1) on proliferation of HUASMCs were determined by MTT assay at different time points (3－96 h) and concentrations (1.09－1090.75 μmol·L^-1). RT-PCR was used determine the effects of EGCG (2.18,21.82,218.15 μmol·L^-1) on expression of IL-6 mRNA in HUASMCs. Double antibody sandwich ELISA was used to determine effects of EGCG (2.18,21.82,218.15 μmol·L^-1) on secretion level of IL-6 in HUASMCs. RESULTS EGCG inhibited the activity of HUASMCs in each detection time point (P<0.01), the inhibiting rate of 3 to 48 h increased with time and reached its peak at 48 h. Compared to inhibiting effects at 48h, no significant difference had been observed in inhibiting effects on proliferation at 72 h and 96 h (P>0.05). Different concentrations of EGCG inhibited the proliferation of HUASMCs (P<0.05 or P<0.01) in a dose dependent manner, with an IC50 of 413.39 μmol·L^-1. EGCG (21.82,218.15 μmol·L^-1) inhibited the expression of IL-6 mRNA of HUASMCs (P<0.01). EGCG also (2.18,21.82,218.15 μmol·L^-1) inhibited the secretion of IL－6 of HUASMCs (P<0.05). CONCLUSION EGCG inhibited the vitro proliferation of HUASMCs. EGCG inhibited the expression of IL-6 mRNA and the secretion of IL-6 in HUASMCs.