达格列净激活自噬并抑制活性氧-核苷酸结合寡聚化结构域样受体蛋白3通路发挥抗动脉粥样硬化的研究

Study of dapagliflozin activating autophagy and inhibiting reactive oxygen species-Nod-like receptor pyrin domain-containing protein 3 pathway in anti-atherosclerosis

目的探究达格列净激活自噬并抑制活性氧(ROS)-核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)通路发挥抗动脉粥样硬化(AS)的作用。方法将30只SD雄性大鼠分为正常组、模型组和实验组。模型组与实验组喂养高脂饲料16周诱导AS。AS建模成功后,实验组予以达格列净(3 mg·kg^(-1))灌胃干预4周,正常组及模型组则灌胃等量的0.9%NaCl干预4周。用酶联免疫吸附试验法检测大鼠血清氧化低密度脂蛋白(ox-LDL)、ROS和白细胞介素1β(IL^(-1)β),用蛋白质印迹法检测腹主动脉的微管相关蛋白轻链3(LC3Ⅱ/Ⅰ)、选择性自噬接头蛋白(p62)和NLRP3蛋白的表达水平。结果实验组、模型组和正常组的血清ox-LDL含量分别为(381.56±108.08)、(640.89±181.06)和(360.19±102.65)pg·mL^(-1),血清ROS含量分别为(8.01±1.39)、(15.47±2.65)和(7.89±1.07)ng·mL^(-1),血清IL^(-1)β含量分别为(84.18±37.90)、(141.44±88.05)和(57.15±14.53)pg·mL^(-1),NLRP3蛋白相对表达水平分别为1.09±0.39、1.55±0.06和0.83±0.16,LC3Ⅱ/Ⅰ蛋白相对表达水平分别为1.03±0.05、0.55±0.02和2.81±0.18,p62蛋白相对表达水平分别为0.97±0.02、1.56±0.07和0.59±0.01。模型组的上述指标与正常组和实验组相比,差异均有统计学意义(均P<0.05)。结论达格列净可能通过激活自噬并抑制ROS-NLRP3通路,从而发挥抗动脉粥样硬化的作用。

Objective To investigate the anti-atherosclerotic(AS)effect of dapagliflozin in activating autophagy and inhibiting reactive oxygen species(ROS)-Nod-like receptor pyrin domain-containing protein 3(NLRP3)pathway.Methods Thirty SD male rats were divided into normal group,model group and experimental group.The model group and experimental group were fed with high-fat diet for 16 weeks to induce AS.After successful modeling of AS,experimental group was intragastrically administered with dapagliflozin(3 mg·kg^(-1))for 4 weeks,while the normal group and model group were given the same amount of 0.9%NaCl for 4 weeks.The serum levels of oxidized low density lipoprotein(ox-LDL),ROS and interleukin-1βin rats(IL-1β)in rats were determined by enzyme-linked immunosorbent assay.Western blot was used to detect the expression levels of microtubule-associated protein light chain 3(LC3 I/I),selective autophagy adaptor protein(p62),NLRP3 proteins in rat abdominal aorta.Results The levels of the ox-LDL in the plasma of experimental group,model group and normal group were(381.56±108.08),(640.89±181.06)and(360.19±102.65)pg·mL^(-1);the levels of the ROS were(8.01±1.39),(15.47±2.65)and(7.89±1.07)ng·mL^(-1)the levels of the IL-1βwere(84.18±37.90),(141.44±88.05)and(57.15±14.53)pg·mL^(-1);the relative expression levels of NLRP3 protein were 1.09±0.39,1.55±0.06 and 0.83±0.16;the relative expression levels of LC3Ⅱ/Ⅰprotein were 1.03±0.05,0.55±0.02 and 2.81±0.18;the relative expression levels of p62 protein were 0.97±0.02,1.56±0.07 and 0.59±0.01.There were significant differences of the above indexes between model group and normal and model groups(all P<0.05).ConclusionDapagliflozin may play an anti-atherosclerotic role by activating autophagy and inhibiting ROS-NLRP3 pathway.