Fe^(2+)对人脐静脉平滑肌细胞增殖及分泌基质金属蛋白酶-2的影响

Effects of Fe^(2+) on proliferation and secretion of MMP-2 in umbilical venous smooth muscle cells

目的探讨Fe2+对高糖预孵育的人脐静脉平滑肌细胞(human umbilical vascular smooth muscle cell,HUSMC)增殖及基质金属蛋白酶-2(MMP-2)分泌的影响。方法取高糖DMEM培养液培养的原代HUSMC(3~5代),分为空白对照组(加DMEM培养液)、A组(100 mg/L Fe2+培养液)、B组(50 mg/L Fe2+培养液)、C组(25 mg/L Fe2+培养液)、D组(12.5 mg/L Fe2+培养液)、E组(6.25 mg/L Fe2+培养液)。干预24 h后,MTT比色法测定细胞增殖,ELISA检测MMP-2含量,激光共聚焦显微镜测定细胞内Ca2+浓度。结果除E组外其他各Fe2+组促血管平滑肌细胞(VSMC)增殖作用均高于空白对照组(P<0.01),A组、B组作用最强;各浓度Fe2+组MMP-2含量均高于空白对照组(P<0.05);A组细胞内Ca2+荧光强度较空白对照组强(P<0.01)。结论Fe2+能明显促进VSMC增殖,并促进分泌MMP-2,升高细胞内Ca2+浓度。

Objective To investigate the effects of Fe^2＋ on proliferation and secretion of MMP-2 in HUVSMC incubated in high glucose DMEM. Methods Primary cultures of HUVSMCs incubated in the high glucose DMEM （Passage 3～5） ,were divided into control group and groups in cubated with Fe^2＋ at five different concentrations（100 mg/L,50 mg/L,25 mg/L,12.5 mg/L,6.25 mg/L）. After 24 h of intervention, HUVSMCs proliferation was assessed with MTT method, MMP-2 secretion was measured with ELISA, and the intracellular Ca2＋ concentration was assessed by laser confocal microscopy. Results Compared with the control group, HUVSMCs proliferation in 12.5～ 100 mg/L Fe^2＋ treatment group was increased （P 〈0.05）;the secretion of MMP-2 was promoted in all Fe^2＋ concentration groups （P 〈0.05） ;and the intracellular Ca^2＋ concentration of HUVSMC was augmented by 100 mg/L Fe^2＋ treatment （P 〈0.01）. Conclusions Fe^2＋ can stimulate the proliferation of VSMC, and increase the intracellular Ca^2＋ as well as the secretion of MMP-2.