加入去甲肾上腺素培养的心肌细胞株H9c2体积变化

Volume change of H9c2 cells treated with norepinephrine

目的观察去甲肾上腺素(NE)对H9c2心肌细胞体积的影响,并探讨其作用机制。方法取处于对数生长期H9c2细胞分为观察组和对照组,分别加入终浓度为2μmol/L的NE、等体积PBS,继续培养48 h。Countstar自动细胞计数仪测算细胞体积,Real Time-PCR法检测细胞心房钠尿肽(ANF)mRNA,免疫荧光法检测细胞氯通道蛋白3(Cl C-3)。结果实验组、对照组细胞体积分别为(2 446.0±193.3)和(1 736.0±265.6)μm3,二者比较,P<0.05。实验组、对照组ANF mRNA分别为1.63±0.14和1.00±0.13,二者比较,P<0.05。实验组、对照组细胞Cl C-3的AOD值分别为0.261±0.062和0.660±0.107,二者比较,P<0.05。结论 NE可导致H9c2细胞体积增大,可能与其上调ANF mRNA表达、抑制Cl C-3蛋白表达有关。

Objective To observe the effect and the mechanism of norepinephrine（NE） on the cell volume of H9c2 cells. Methods Cells in the logarithmic growth phase were divided into two groups： the control group and experimental group. Cells in the experimental group were treated with 2 μmol/L NE for 48 h,and the cells in the control group were incubated with equal volume of PBS for 48 h. The volume of H9c2 cells was detected by Countstar automated cell counter.The real-time PCR was used to measured the relative atrial natriuretic factor（ANF） mRNA level of the cells. The expression of chloride channel protein 3（Cl C-3） was measured by immunofluorescence. Results The volume of H9c2 cells in the control group and experimental group were（1736. 0 ± 265. 6） and（2446 ± 193. 3） μm3,respectively; the levels of ANF mRNA in the control group and experimental group were 1. 00 ± 0. 13 and 1. 63 ± 0. 14,respectively; the expression levels of Cl C-3 in the control group and experimental group were 0. 660 ± 0. 107 and 0. 261 ± 0. 062,respectively; significant differences were found between these two groups（all P〈0. 05）. Conclusion NE increases the volume of H9c2 cells by up-regulating the expression of ANF and down-regulating the expression of Cl C-3.