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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.
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.
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).
AA can alleviate HCY induced apoptosis of cardiac microvascular endothelial cells and promote vascular regeneration by inhibiting TLR4/MyD88/NF-κB pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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