血清炎症因子在糖尿病性心肌病患者中的表达及其对心肌细胞线粒体功能的影响

Expression of serum inflammatory cytokines in patients with diabetic cardiomyopathy and their effects on mitochondrial function of cardiomyocytes

  • 摘要:
    目的 探讨炎症因子白细胞介素(IL)-1β和IL-18在糖尿病性心肌病(DCM)中的表达水平及其对心肌细胞线粒体功能的影响。
    方法 采集104例DCM患者(DCM组)和90例健康志愿者(健康对照组)的外周血血清,检测2组血清IL-1β和IL-18浓度。培养人心肌细胞系AC16,将AC16细胞分为对照组、IL-1β组(加入10 ng/mL的重组人IL-1β)和IL-18组(加入10 ng/mL的重组人IL-18)。采用CCK-8法检测细胞增殖活性;分别采用JC-1染色法和钙黄绿素染色法检测心肌细胞线粒体膜电位和膜通透性;采用Western blot检测细胞凋亡相关蛋白Bax、Bcl-2和Cleaved-Caspase-3(C-Casp3)的表达水平。
    结果 与健康对照组相比,DCM组血清IL-1β和IL-18浓度升高,差异有统计学意义(P < 0.05)。与对照组比较,AC16细胞加入IL-1β或IL-18培养48、72 h后,AC16细胞增殖活性下降,差异有统计学意义(P < 0.05);与对照组相比,IL-1β组和IL-18组线粒体膜电势下降的细胞比率增加,差异有统计学意义(P < 0.001);IL-1β组、IL-18组钙黄绿素平均荧光强度均高于对照组,差异有统计学意义(P < 0.001)。与对照组相比,IL-1β组和IL-18组细胞Bax/Bcl-2、C-Casp3/GAPDH比值升高,差异有统计学意义(P < 0.01或P < 0.001)。
    结论  IL-1β和IL-18在DCM患者外周血中表达水平升高,并通过抑制线粒体活性影响其心肌细胞功能。

     

    Abstract:
    Objective  To investigate the expression levels of inflammatory cytokines interleukin (IL)-1β and IL-18 in diabetic cardiomyopathy (DCM) and their effects on mitochondrial function of cardiomyocytes.
    Methods  The peripheral blood serum of 104 DCM patients (DCM group) and 90 healthy volunteers (healthy control group) were collected, and the serum levels of IL-1β and IL-18 in the two groups were detected. Human myocardial cell line AC16 was cultured and divided into control group, IL-1β group (adding 10 ng/mL recombinant human IL-1β) and IL-18 group (adding 10 ng/mL recombinant human IL-18). Cell proliferation activity was detected by CCK-8 assay; mitochondrial membrane potential and permeability were detected by JC-1 staining and calcein staining, respectively; the expression levels of apoptosis-related proteins Bax, Bcl-2 and Cleaved Caspase-3 (C-Casp3) were detected by western blot.
    Results  Compared with the healthy control group, serum IL-1β and IL-18 concentrations in the DCM group were increased, and the difference was statistically significant (P < 0.05). Compared with the control group, the proliferating activity of AC16 cells was significantly decreased after incubation with IL-1β or IL-18 for 48 and 72 h (P < 0.05); compared with the control group, the percentage of cells with decreased mitochondrial membrane potential in the IL-1β and IL-18 groups was significantly increased (P < 0.001); the average fluorescence intensity of calcein in the IL-1β group and IL-18 group was significantly higher than that in the control group (P < 0.001). Compared with the control group, the Bax/Bcl-2 and C-Casp3/GAPDH in the IL-1β and IL-18 groups were significantly increased (P < 0.01 or P < 0.001).
    Conclusion  The expression levels of IL-1β and IL-18 in peripheral blood of DCM patients are increased, and the function of cardiomyocytes is affected by inhibiting mitochondrial activity.

     

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