Anti-atherosclerosis drug efficacy evaluation method based on inflammatory responses
A technology for atherosclerosis and drug efficacy evaluation, which is applied in the fields of compound screening, material inspection products, and detection of programmed cell death.
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Embodiment 1
[0089] 1. Method for establishing EC-MC-SMC co-culture system:
[0090] 1.1 First plant SMC (HASMC) at 1×105 / cm2 (volume 50 μl) on the outside of the PE membrane of Millicellinsert, turn over after 8 hours for the cells to adhere to the wall, and plant EC (HUAEC) at 1×105 / cm2 on the inner side of the membrane, After co-culture for 6 days, mononuclear cells THP-1 were inoculated at 1×105 / cm2, and oxLDL100μg / ml, IL-1β10ng / ml and test drug were added to culture for 24h. Pharmacodynamic experiments were carried out in this system.
[0091] 1. After 224 hours, use the tip of a pipette to gently absorb the medium in the Millicellinsert, repeat 3 times, and then absorb the supernatant (about 400 μl) into a 1ml EP tube and centrifuge at 2500rpm at 4°C for 10min. Take the supernatant and freeze it. Used to detect EC layer MCP-1, TNFα.
[0092] 1.3 The pellet is the pellet of mononuclear THP-1 cells, which is resuspended in 1ml PBS, and CD36 on the surface of monocytes is detected by...
Embodiment 2
[0104] 1 unknown drug experiment
[0105] Tanshinone IIA (Tan IIA)
[0106] The EC-SMC-MC co-culture group was used as a control, and atorvastatin was used as a positive control to detect according to the method of Example 1, and the detection results were as follows:
[0107] 1.1 The effect of tanshinone IIA on the secretion of MCP-1 and TNFα from EC cells The results showed that EC-SMC-MC co-cultured and stimulated with oxLDL and IL-1, ICAM-1 on the surface of EC, and TNFα, MCP- 1 was significantly increased. Tanshinone IIA can reduce the expression of ICAM-1 on the surface of EC, and the content of TNFα and MCP-1 in the co-culture supernatant, and there are significant differences compared with the model group (P<0.05 or P<0.01).
[0108] Table 5 Effect of Tanshinone IIA on the secretion of MCP-1 and TNFα from EC cells
[0109]
[0110] 1.2 The effect of tanshinone IIA on the expression of CD36 on the surface of monocytes The results showed that the expression of CD36...
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