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目的:探讨阿司匹林对高糖和高胰岛素诱导的大鼠血管平滑肌细胞(vasculersmoothmusclecells,VSMCs)内诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)含量变化的影响,以及高糖、高胰岛素状态下细胞内iNOS含量变化与上清液中NO含量变化的相关性。方法:组织贴块法原代培养大鼠胸主动脉VSMCs,用阿司匹林干预高糖和高胰岛素诱导后的VSMCs。实验设正常对照组(CON组)、高糖组(HG组)、高胰岛素组(HI组)、高糖和高胰岛素组(HGI组)、阿司匹林2.50mmol/L+HG组(AHG组)、阿司匹林2.50 mmol/L+HI组(AHI组)、阿司匹林2.50 mmol/L+HGI组(AHGI组),每组样本数为6。应用大鼠iNOS ELISA试剂盒检测细胞内iNOS的变化情况(OD值),NO试剂盒检测培养基上清液中NO含量,并分析高糖、高胰岛素状态下两者的相关性。结果:高糖、高胰岛素均能使VSMCs内iNOS含量减低(P<0.05),高糖和高胰岛素联合能使VSMCs内iNOS含量明显减低(P<0.01);阿司匹林能使高糖、高胰岛素刺激后VSMCs内iNOS含量升高(P<0.05),并能显著提高高糖和高胰岛素联合刺激后VSMCs内iNOS含量(P<0.01)。高糖、高胰岛素、高糖和高胰岛素联合均能使培养基上清液中NO含量减低(P<0.05);阿司匹林能提升上述三者刺激后上清液中NO含量(P<0.05)。HGI组中iNOS的降低与上清液中NO的降低、AHGI组iNOS的升高与上清液中NO的升高并不表现出相关性(r=0.778,r=0.632,均P>0.05)。结论:阿司匹林能提高高糖和高胰岛素联合刺激后VSMCs内iNOS的含量,并能提高培养液中NO的含量,但细胞内iNOS含量可能不是决定培养液中NO含量的唯一因素。
AIM: To investigate the effects of aspirin on changes of inducible nitric oxide synthase (iNOS) in rat vascular smooth muscle cells (VSMCs) induced by high glucose and high insulin, and to investigate the effects of high glucose and insulin status Correlation between changes of iNOS content and NO content in supernatant of inferior cells. Methods: VSMCs of thoracic aorta were primarily cultured by tissue patch method. Aspirin was used to interfere with VSMCs induced by high glucose and high insulin. The experimental groups were divided into normal control group (CON group), high glucose group (HG group), high insulin group (HI group), high glucose and high insulin group (HGI group), aspirin 2.50mmol / L + HG group Aspirin 2.50 mmol / L + HI group (AHI group), aspirin 2.50 mmol / L + HGI group (AHGI group), the number of samples in each group was 6. The changes of intracellular iNOS (OD value) were detected by rat iNOS ELISA kit. The content of nitric oxide (NO) in the supernatant was detected by NO kit, and the correlation between high glucose and high insulin was analyzed. Results: Both high glucose and high insulin could reduce the iNOS content in VSMCs (P <0.05). The combination of high glucose and high insulin could significantly reduce the content of iNOS in VSMCs (P <0.01). Aspirin could make high glucose and insulin stimulation The content of iNOS in VSMCs was increased (P <0.05), and the content of iNOS in VSMCs after high glucose and high insulin combination stimulation was significantly increased (P <0.01). The combination of high glucose, high insulin, high glucose and high insulin reduced the NO content in the supernatant (P <0.05). Aspirin increased the NO content in the supernatant (P <0.05). The decrease of iNOS in HGI group and the decrease of NO in supernatant showed no correlation between the increase of iNOS in AHGI group and the increase of NO in supernatant (r = 0.778, r = 0.632, all P> 0.05) . CONCLUSIONS: Aspirin can increase the content of iNOS in VSMCs induced by high glucose and high insulin, and increase the content of NO in culture medium. However, the intracellular iNOS content may not be the only factor that determines the content of NO in culture medium.