摘要
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 investigate the changes of α1-adrenergic receptor subtypes on gene level during inflammatory response. Methods Primary cultures of vascular smooth muscle cells （VSMC） allocated into the following groups： medium （control）, tyrosine （Tyr, 250 μmol/L）, 3-nitrotyrosine（3-NT, 250μmol/L）, S-nitroso-N-acetylpenicillamine （SNP, 500μmol/L）, a mixture of TNF-α and IL-1β （Cyto,TNF-α 100 ng/ml, IL-1β 50 ng/ml）, Nω-nitro - L-arginine methyl ester with a mixture of these cytokines（Cytoq-L-NAME, 5 mmol/L）, melatonin with a mixture of these cytokines （Cytoq-MLT, I mmol/L）. Total RNA from VSMC was extracted and α1A-AR, α1B-AR and α1B-AR mRNA as well as β-actin mRNA were measured by reverse transcription polymerase chain reaction（RT-PCR）. The mRNA values were related to signals obtained for β-actin mRNA. Results Incubation of cultured VSMC with 3-NT resulted in diminished α1B-AR and α1D-AR gene expression,and mRNA level of α1A-AR, α1B-AR and α1D-AR was decreased after incubation with SNP. Incubation of the cells with the mixture of cytokines also caused down regulation of α1A-AR and α1D-AR mRNA,whereas L-NAME or melatonin attenuated these down regulation. Conclusion NO and its metabolic derivant 3-NT significantly attenuate α1-ARs gene expression in VSMC during inflammatory response, which may be one of possible pathophysiological mechanisms of vascular hyporeactivity during septic shock. The use of antioxidants and depression of NO synthesis may partly reduce this pathological response.
出处
《江苏医药》
CAS
CSCD
北大核心
2005年第8期596-598,共3页
Jiangsu Medical Journal
基金
国家教委归国人员基金资助(00GJ2)
铁道部科基金资助(J2000Z087)
