Abstract
Objective To evaluate the effects of centella asiatica acid(AA)on the apoptosis of cardiac microvascular endothelial cells(CMECs)and vascular regeneration by modulating the Toll-like receptor 4/myeloid differentiation primary response 88/nuclear factor kappa B/nucleotide-binding oligomerization domain-like receptor protein 3(TLR4/MyD88/NF-κB/NLRP3)pathway.Methods The homocysteine(HCY)-induced rat CMECs were used to simulate and construct a coronary heart disease injury cell model,and the success of model construction was verified using the cell counting kit-8(CCK8)method.The cells were divided into control group(normal culture),model group(modeling),centella asiatica acid group(modeling+10 μmol·L-1 AA)and combined group(modeling+1 μg·mL-1 lipopolysaccharide+10 μmol·L-1 AA).Cell viability was detected using CCK8,apoptosis was assessed by flow cytometry,and the relative expression levels of apoptosis-related proteins,pathway-related proteins and vascular regeneration-related proteins were measured by Western blot.Results The cell viability of control group,model group,asiatica acid group and combination group were 0.92±0.01,0.61±0,0.75±0.01 and 0.50±0.02,respectively;the apoptosis rates were(4.96±0.53)%,(24.98±1.68)%,(18.22±0.96)%and(34.17±0.49)%,respectively;the relative protein expression levels of B-cell lymphoma/leukemia 2-associated X protein(Bax)were 0.12±0.01,0.63±0,0.42±0.02 and 0.96±0.01,respectively;the relative expression levels of Bcl-2 protein were 0.99±0.02,0.34±0.01,0.69±0.01 and 0.10±0.01,respectively;the relative expression levels of TLR4 protein were 0.19±0.01,0.65±0,0.41±0.01 and 0.84±0.01,respectively;the relative expression levels of Myd88 protein were 0.29±0.02,0.77±0,0.61±0.01 and 0.95±0.02,respectively;the relative expression levels of NF-κ B protein were 0.22±0.01,0.82±0.02,0.53±0.02 and 0.84±0.02,respectively;the relative expression levels of NLRP3 protein were 0.17±0.02,0.75±0.03,0.50±0.01 and 0.98±0.01,respectively;the relative expression levels of vascular endothelial growth factor(VEGF)protein were 0.95±0.01,0.44±0.02,0.68±0.02 and 0.23±0.01,respectively;the relative expression levels of platelet endothelial cell adhesion molecule-1(CD31)protein were 1.01±0.01,0.44±0.02,0.58±0 and 0.38±0.01,respectively.The above indicators of model group were compared with control group,the above indicators of the oxalic acid group were compared with the model group,and the above indicators of the combination therapy group were compared with the asiatica acid group,the differences were all statistically significant(all P<0.01).Conclusion AA can alleviate HCY induced apoptosis of cardiac microvascular endothelial cells and promote vascular regeneration by inhibiting TLR4/MyD88/NF-κB pathway.关键词
积雪草酸/冠心病/Toll样受体4/髓样分化因子88/核因子κB/NOD样受体蛋白3/凋亡/血管生成Key words
asiatic acid/coronary heart disease/Toll-like receptor 4/myeloid differentiation primary response 88/nuclear factor kappa B/nucleotide-binding oligomerization domain-like receptor protein 3 pathway/apoptosis/angiogenesis分类
医药卫生