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目的探讨缺血性脑血管病(ICVD)患者外周血细胞自噬因子的表达意义。方法选取浙江省湖州市第三人民医院急诊内科收治的96例ICVD患者为研究对象,根据颈动脉斑块分级分为Ⅰ、Ⅱ、Ⅲ3组,同时以30例同期健康志愿者为对照,进行同步研究。取空腹静脉血,在透射电子显微镜和荧光显微镜下观察自噬小体;采用蛋白免疫印迹法测定自噬因子Beclin-1、LC-Ⅱ/LC-Ⅰ及p62的表达水平;采用酶联免疫吸附试剂盒检测血清氧化低密度脂蛋白(ox-LDL)、脂蛋白相关磷脂酶A2(Lp-PLA2)、肿瘤坏死因子α(TNF-α)与白细胞介素6(IL-6)的血清水平。用SPSS 16.0统计软件进行数据分析,多组间比较用方差分析,进一步两两比较用SNK检验;计数资料的组间比较用χ2检验;相关性分析用简单线性回归分析。结果对照组、Ⅰ级组、Ⅱ级组、Ⅲ级组自噬小体荧光强度逐渐增强,Beclin1、LC-3Ⅱ/LC-3Ⅰ表达逐渐升高,p62表达逐渐降低,组间差异均有统计学意义(P<0.05);对照组、Ⅰ级组、Ⅱ级组、Ⅲ级组的血清ox-LDL、Lp-PLA2、TNF-α、IL-6水平逐渐升高,组间差异均有统计学意义(P<0.05);ICVD患者的LC-3Ⅱ/LC-3Ⅰ表达与ox-LDL、Lp-PLA2、TNF-α及IL-6水平均呈正相关性,Beclin1表达与ox-LDL、Lp-PLA2水平呈正相关性,p62表达与Lp-PLA2、TNF-α、IL-6水平呈负相关性(P<0.05)。结论 ICVD患者外周血细胞自噬因子水平关系到患者斑块的炎性反应和稳定性,可能是其介导ICVD病理机制的重要途径之一。
Objective To investigate the significance of autophagy in peripheral blood cells in patients with ischemic cerebrovascular disease (ICVD). Methods Ninety-six ICVD patients admitted to the Third People’s Hospital of Huzhou City, Zhejiang Province, China were selected as study objects. According to the classification of carotid plaques, they were divided into Ⅰ, Ⅱ and Ⅲ groups. At the same time, 30 healthy volunteers the study. Fasting venous blood was taken and autophagic bodies were observed under transmission electron microscope and fluorescence microscope. The expression of autophagy factors Beclin-1, LC-Ⅱ / LC-Ⅰ and p62 were detected by Western blotting. Serum levels of ox-LDL, Lp-PLA2, TNF-α and IL-6 were detected by kit. SPSS 16.0 statistical software was used for data analysis. ANOVA was used to compare among groups. SNK test was used to compare the data between groups. Χ2 test was used to compare the data between groups. Correlation analysis was performed by simple linear regression analysis. Results The fluorescence intensity of autophagosomes in control group, Ⅰ group, Ⅱ group and Ⅲ group gradually increased, the expression of Beclin1, LC-3Ⅱ / LC-3Ⅰwere gradually increased and the expression of p62 was gradually decreased with statistical difference (P <0.05). The levels of serum ox-LDL, Lp-PLA2, TNF-α and IL-6 in control group, Ⅰ group, Ⅱ group and Ⅲ group increased gradually with statistical significance (P <0.05). The expression of LC-3Ⅱ / LC-3Ⅰwas positively correlated with the level of ox-LDL, Lp-PLA2, TNF-α and IL-6 in ICVD patients There was a positive correlation between p62 expression and Lp-PLA2, TNF-α and IL-6 levels (P <0.05). Conclusion The level of autophagy in peripheral blood of patients with ICVD is related to the inflammatory response and the stability of plaques in patients with ICVD. It may be one of the important pathways that mediate ICVD pathological changes.