The Effects Of Chrysanthemum Indicum Extract On The Expressions Of TIPE2 And PCNP In Human Vascular Endothelial Cell

Background Atherosclerosis is the basic pathological change of cardiovascular and cerebrovascular diseases which are of high morbility and fatality. With the improvement of the level of industrialization and living conditions, the disease grows more popular and younger in age. The pathogenesis of atherosclerosis has yet not been fully elucidated. The most widely accepted hypothesis is the inflammatory reaction theory, which says that atherosclerosis is a chronic inflammatory disease involving the interaction of blood components and arterial wall components. A number of studies have shown that a variety of inflammatory mediators and growth factors(cytokines) participate in the formation of atherosclerotic plaques. However the clinical treatments based on the latest researches are still limited. The application of basic medical science into clinical practice is not always successful. The incidence of myocardial infarction is still as high as 20% within 3 years in patients suffering from acute coronary syndrome, even if they get the existing best drug treatment and interventional therapy. It is of great imminence to find an effective and economical method to prevent and slow the progression of atherosclerosis. The damagement to the vascular endothelial cells is the blasting fuse of atherosclerosis. TIPE2 is a member of the TNFAIP8 family, which plays a crucial role in the maintenance of immunehomeostasis. TIPE2 negatively regulates the inflammatory response through the T cell receptor and Toll-like receptor signaling pathway. PCNP is a nuclear protein involved in cell cycle regulation. It is also an effector of signaling pathway related to cell proliferation. In our country, chrysanthemum has been used as a natural herb to prevent and treat diseases for thousands of years. Modern medical researches have already proved that chrysanthemum indicum extract has the power of reducing inflammatory mediators, inhibiting tumor cell proliferation, and preventing atherosclerosis. Nowadays chrysanthemum indicum extract has been used as an antioxidant and anti-inflammatory agent, which possesses cardiovascular protective effects. But it’s a pity that the underlying molecular mechanism is still mysterious. As we know, there exists inflammation reaction and cell proliferation throughout the atherosclerotic process. So we would like to explore whether there are some relationships among chrysanthemum or its extract, TIPE2, PCNP and atherosclerosis.Aims This study is to investigate the effects of CIE on the expressions of TIPE2 and PCNP in human aortic endothelial cell(HAEC) and human coronary artery endothelial cell(HCAEC).Methods First we determinated that CIE exerted no toxic effects on cultured HAEC and HCAEC by MTT assays. Then the cells were cultured for 24 h,48h,72 h,respectively,in mediums added with proper different concentrations of CIE. Then total cellular m RNA and total protein were extracted. We tested the TIPE2 and PCNP expression changes in gene and protein level by Real-time PCR and Western Blot.Results 1.Real-time PCR analysis suggested that treatment with CIE significantly increased the expression of m RNA of TIPE2 gene than the control group. But CIE exerted no obvious influence on the expression of PCNP. 2.Western Blot analysis showed that the expression of protein of TIPE2 gene is much higher than that of the experimental group. There is a positive correlation between the increase in the protein expression of TIPE2 and the concentration of CIE,with statistically significant difference. But CIE exerted no obvious influence on the expression of PCNP.Conclusions CIE has no toxic effects on the growth of HAEC and HCAEC. CIE could increase the m RNA and protein expression of TIPE2 gene. CIE may play its pharmacological effects by influencing the expression of TIPE2 while CIE exerted no obvious influence on PCNP.