| Background and Objective Atherosclerosis（AS）is the typical pathological basis of coronary heart disease.Inflammation and dyslipidemia underlie the pathological basis of AS.Clinical studies have confirmed that there is still residual risk of atherosclerotic cardiovascular diseases（ASCVD）even after intense reduction of LDL.Some of this residual risk can be explained by inflammation as anti-inflammatory therapy is effective in improving the efficacy of LDL inhibitor therapy.Nod-like receptor pyrin domain-containing protein 3（NLRP3）inflammasome is believed to be a link between lipid metabolism and inflammation,as cholesterol crystallization and oxidized low-density lipoprotein（ox-LDL）can activate NLRP3 inflammasome.Research has also found that reactive oxygen species（ROS）can promote the conversion of low-density lipoprotein（LDL）into ox-LDL,further leading to ROS secretion and activating NLRP3 inflammasomes and downstream cytokine release.Autophagy can inhibit the activation of NLRP3 and subsequent cytokines by clearing the accumulation of ROS.In this study,we constructed an AS rat model to investigate whether autophagy inhibits ROS-NLRP3 pathway to improve AS and whether dapagliflozin inhibits the development of AS by activating autophagy and inhibiting ROS-NLRP3 pathway,and explore the possible mechanism of daggligin to improve AS at the animal level.Methods Forty male SD rats were divided into a blank control group（n=10）and an AS model construction group（n=30）.The AS model construction group was given vitamin D3 by gavage（the first 3 days）and fed with high-fat feed for 16 weeks to induce AS the formation of atheroscloma lesions.After successful modeling of AS,it was divided into a model group,a rapamycin group,and daggliflozin group.The rapamycin group and daggliflozin group were intragastrically administered by rapamycin(1 mg·kg-1)and daggliflozin(3 mg·kg-1)for 4 weeks respectively,while the blank control group and model group were given the same amount of0.9%Na Cl for 4 weeks.At the end of drug intervention,aortic oil red O staining was observed the area of AS lesions;Hematoxylin eosin staining（HE）was used to observe foam cells;The serum levels of rat in GLU、TC、TG、HDL-C、LDL-C were determined by colorimetric method.The serum levels of rat in oxidized low density lipoprotein（ox-LDL）,ROS and IL-1β in rats were determined by enzyme-linked immunosorbent assay.Western blot was used to detect the expression level of microtubule-associated protein light chain 3（LC3）,selective autophagy adaptor protein（p62）,NLRP3 proteins in rat abdominal aorta.Results⑴Weight Compared with the successful construction of the AS model,after 4 weeks of drug intervention,the weight of the two groups（rapamycin group and daggliflozin group）significantly reduced,with a statistically significant difference（all P<0.05）.⑵Serum indicators The levels of the LDL-C in the plasma of the blank normal control group,model group,rapamycin group and dapagliflozin group were（2.67±1.94）,（19.75±4.86）,（9.18±5.16）and（9.19±1.46）mmol·L-1;the levels of the ox LDL in the plasma of the blank normal control group,model group,rapamycin group,and dapagliflozin group were（375.06±115.43）（586.39±223.70）,（358.17±107.32）and（400.45±129.21）pg·mL-1;the levels of the ROS in the plasma of the blank normal control group,model group,rapamycin group,and dapagliflozin group were（7.89±1.07）,（15.47±2.65）,（8.45±2.20）and（8.17±1.52）ng·mL-1;the levels of the IL-1βin the plasma of the blank normal control group,model group,rapamycin group,and dapagliflozin group were（62.97±18.21）,（171.22±80.2）,（65.50±18.55）和（85.70±38.82）pg·mL-1;There were significant differences of the aboves between model group and blank normal control group and rapamycin group and dapagliflozin group（all P<0.05）.⑶Histomorphology After the end of drug intervention,gross oil red staining of the aorta of rats showed that the plaque area of the model group was significantly increased,compared to the blank control group.Compared with the model group,the plaque area in the rapamycin and daggliflozin groups was significantly reduced,and there was no significant difference between the two groups.HE staining showed that the structure of aortic arch in the blank control group was clear and the morphology was normal.The intima of aortic tissue in the model group was thickened,and foam cells were visible.The drug group significantly alleviated the lesion.⑷Tissue protein expression The relative expressions levels of NLRP3protein were 0.52±0.01,1.58±0.03,0.80±0.01 and 0.84±0.05;the relative expressions levels of LC3-Ⅱ/LC3-Ⅰprotein were 2.69±0.19,0.55±0.25,0.74±0.25 and 0.99±0.17;the relative expression levels of p62 protein were 0.72±0.14,1.41±0.14,0.91±0.01 and 0.79±0.01.There were significant differences of the aboves between model group and blank normal control group and rapamycin group and dapagliflozin group（all P<0.05）.Conclusion Sodium-glucose cotransporter 2 inhibitor dapagliflozin may play an anti-atherosclerosis by activating autophagy and inhibiting ROS-NLRP3pathway. |
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