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目的观察炎症反应时血管平滑肌细胞α1肾上腺素受体基因转录水平的改变。方法原代培养大鼠胸主动脉血管平滑肌细胞(VSMC)分为对照组(含10%FBS的DMEM培养液)、酪氨酸组(Tyr,250μmol/L)、3硝基酪氨酸组(3NT,250μmol/L)、硝普钠组(SNP,500μmol/L)、炎性细胞因子组(Cyto,TNFα100ng/ml+IL1β50ng/ml)、炎性细胞因子组+NωnitroLargininemethylester(Cyto+LNAME,TNFα100ng/ml+IL1β50ng/ml+LNAME5mmol/L)和N乙酰5甲氧基色胺+炎性细胞因子组(Cyto+MLT,TNFα100ng/ml+IL1β50ng/ml+MLT1mmol/L),每组重复6次。上述各组细胞干预12h后,提取各组细胞的总RNA,逆转录聚合酶链反应(RTPCR)法测定α1AAR、α1BAR、α1DARmRNA的表达。结果与对照组相比,SNP组和Cyto组血管平滑肌细胞细胞α1AAR、α1DARmRNA的表达显著下降(P<0.05),Cyto+LNAME组、Cyto+MLT组上述各受体亚型mRNA的表达比Cyto组显著升高(P<0.01)。与对照组相比,SNP组、3NT组α1BAR、α1DARmRNA表达明显降低(P<0.05)。结论NO及NO衍生物3NT均显著抑制了血管平滑肌细胞细胞α1肾上腺素受体mRNA的表达,可能为感染性休克时血管低反应性的病理机制之一。抑制NO的产生和抗氧化剂治疗可减轻这种病理反应。
Objective To observe the changes of α1 adrenergic receptor gene transcription in vascular smooth muscle cells during inflammatory reaction. Methods Primary cultured rat aortic vascular smooth muscle cells (VSMCs) were divided into control group (DMEM containing 10% FBS), tyrosine group (Tyr, 250μmol / L) and 3-nitrotyrosine group (Cyto, TNFα 100ng / ml + IL1β50ng / ml), inflammatory cytokines + NωnitroLargininemethylester (Cyto + LNAME, TNFα100ng / ml + IL1β50ng / ml + LNAME5mmol / L) and N-acetyltryptamine serotonin + inflammatory cytokines (Cyto + MLT, TNFα100ng / ml + IL1β50ng / ml + MLT1mmol / L). After 12 hours of intervention, the total RNA of each group was extracted and the expression of α1AAR, α1BAR and α1DAR mRNA was determined by reverse transcription polymerase chain reaction (RTPCR). Results Compared with the control group, the expressions of α1AR and α1DARmRNA in the vascular smooth muscle cells of the SNP group and the Cyto group were significantly decreased (P <0.05). Compared with the Cyto + LNAME group and Cyto + MLT group, Significantly increased (P <0.01). Compared with the control group, the expressions of α1BAR and α1DARmRNA in SNP group and 3NT group were significantly decreased (P <0.05). Conclusion Both NO and NO derivatives 3NT can significantly inhibit the expression of α1-adrenoceptor mRNA in vascular smooth muscle cells, which may be one of the pathological mechanisms of vascular hyporesponsiveness during septic shock. Inhibition of NO production and antioxidant therapy can reduce this pathological response.