To Observe the Effect of API1034 on CD40 of Mouse Peritoneal Macrophages
|School||Huazhong University of Science and Technology|
|Keywords||AS macrophage CD40/CD40L API0134 immunohistochemistry RT-PCR|
Atherosclerosis is currently considered to be an inflammatory and immunomodulatory disease. It has been demonstrated that inflammation-signal transduction of CD40-CD40L plays an important role on atherosclerosis. the CD40/CD40L co-stimulatory can adjust the expression of inflammation cytokine, chrematistic-factor and adhesion molecule, start the demic immune response, promote the development of atherosclerotic plaque, participate in atherosclerotic plaque rupture and stimulate thrombosis. Disrupting the CD40/CD40L co-stimulatory pathway reduces atherosclerosis and induces a stable atherosclerotic plaque phenotype that is low in inflammation and high in fibrosis. Therefore, inhibition of the CD40/CD40L interaction pathway is an attractive therapeutic target to reduce clinical complications of atherosclerosis. The prophase study has been confirmed that API0134 can play a significant role on inhibition of AS. Interfering of the CD40/CD40L co-stimulatory pathway ,inhibiting the immune response and inflammatory reaction, may be its mechanism of action .This study is to observe the effect of API0134 on expression level of CD40 of Mouse Peritoneal Macrophages, and judge whether API0134 interfere the CD40/CD40L co-stimulatory pathway or not? Objective: The cultured Macrophages cells put in a medium of the same concentration 30μg/ml ox-LDL and different concentration 10μɡ/ml, 50μg/ml, 100μg/ml API0134, to detect the expression level of CD40/CD40L of Mouse Peritoneal Macrophages after having been cultured 24 hours. Therefore we can judge whether API0134 may play a intervention on expression level of CD40/CD40L.Methods: Isolation and culture of Mouse Peritoneal Macrophages is the first. The experiment groups are designed as control group, without API0134, and 10μɡ/ ml ,50μg/ml, 100μg/ml API0134. All are put in 10% calf bovine serum.。After 24 hours, we can detect the expression level of CD40/CD40L of Mouse Peritoneal Macrophages by immunohistochemistry and RT-PCR. According to the blank , the OX-LDL , the OX-LDL+ API0134,we detect respectively the expression level of CD40/CD40L of Mouse Peritoneal Macrophages by the kind of immunohistochemistry. The expression level of CD40/CD40L of mouse Peritoneal Macrophages is represented by the intensity of immunofluorescence.( IMF)Result: by the kind of RT-PCR, we draw a conclusion API0134 inhibit the expression level of CD40/CD40L of Mouse Peritoneal Macrophages. Comparing to the blank, the 100μg/ml API0134 possess statistical significance, (p﹤0.05) the intensity of immunofluorescence (IMF).of OX-LDL is the most powerful, which the blank, OX-LDL+ API0134 is very lower than OX-LDL.Conclusions: API0134 inhibit the expression level of CD40/CD40L of Mouse Peritoneal Macrophages。It indicates when the concentration of API0134 get somehow level, the API0134 can play role.It can be a new kind of treatment of AS by the inhibition of inflammation-signal transduction of CD40/CD40L.