3-硝基酪氨酸对大鼠血管平滑肌细胞肾上腺素能受体α_(1D)mRNA表达的影响

Effect of nitrotyrosine on the expression of α1D-adrenoceptor mRNA in vascular smooth muscle of rat

目的 探讨3-硝基酪氨酸(3-NT)对大鼠血管平滑肌细胞α1D-肾上腺紊能受体(α1D-AR)mRNA表达的影响。方法 取原代培养的大鼠血管平滑肌细胞。以0、1、10、100、200 μmol／L 3-NT干预24 h,另取培养好的细胞,以100μmol／L 3-NT分别干预0、12、24、48、72 h,逆转录-聚合酶链反应检测各浓度点和时间点α1D-AR mRNA的表达。结果 与0 μmol／L 3-NT相比,1、10 μmol／L 3-NT对α1D-ARmRNA表达的差异无统计学意义,而100、200 μmol／L 3-NT使α1D-AR mRNA的表达降低(P<0.05)。以100 μmol／L 3-NT分别干预不同时间后,与干预Oh比较,干预12 h时α1D-AR mRNA的表达量无变化,干预24、48、72 h时α1D-AR mRNA的表达降低(P<0.05)。结论 3-NT可能通过抑制α1D-AR mRNA的表达。

Objective To investigate the effect of nitrotyrosine (3-NT) on the expression of α1D adrenoceptor (α1D-AR) mRNA in cultured vascular smooth muscle cells (SMCS) .Methods SMCS were obtained from the tunica media of thoracic aorta of 1 month old SD rats and cultured in DMEM medium. The experiment consisted of two parts. In part Ⅰ SMCS were incubated with 0,1, 10, 100 or 200 μmol·L-1 3-NT for 24 h and in part Ⅱ SMCS were incubated with 100 μmol·L-1 3-NT forO,12, 24, 48 or 72 h. The total RNA was isolated by using Trizol reagent. The expression of α1D-AR mRNA was determined by RT-PCR and agarose gel electrophoresis. Results In part I incubation with 1 and 10 μmol·L- 3-NT for 24 h had no significant effect on the expression of α1D-AR mRNA while incubation with 100 or 200 μmol·L-1 3-NT for 24 h decreased the expression of α1D-AR mRNA compared with 0μmol·L-1 3-NT (P < 0.05). In part ⅡI incubation with 100 μmol·L-1 3-NT for 12 h did not significantly affect α1D-AR mRNA expression compared with 100 μmol·L-1 3-NT for 0 h, whereas incubation with 100 μmol· L-1 3-NT for 24, 48 and 72 h decreased the expression of α1D-AR mRNA ( P < 0.05). Conclusion Our results suggest that 3-NT inhibits the expression of α1D-AR mRNA in SMCS.