Research Of Ten Kinds Chinese Medicine Monomer Composition Of Antiatherosclerosis Screening And Mechanism

Atherosclerosis (AS) is a kind of vascular disease which is chronic and inflammatory."inflammation hypothesis" and "damage-response doctrine "etc have become one of the prevailing doctrine of AS pathogenesis. In the inflammatory signal transduction, NFκB, MAPK, JAK-STAT and T cell costimulatory mediate costimulatory signaling pathway has an important meaning.We choosed the NFκB, extracellular regulated protein kinases (ERK) and c-Jun N-terminal kinase (JNK) which belongs mitogen-activated protein kinase (MAPK) pathways as drug screening target point constructed three typical inflammatory signaling pathways, with which simulation artery atherosclerotic lesions in inflammatory pathways which induced AS.With this high-flux screening model choose the active drug to treat Atheromatosis.In the study, nuclear transcription factor-kappa B (NF-kappa B), extracellular regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK)(they may be triggered signaling pathway transduction) are used as signal target, to construct three kinds inflammatory signaling pathways of Atheromatosis, then transfer recombinant plasmid of NFκB, ERK, JNK, target gene fragment into the characteristic cells-umbilical vein endothelial cells (HUVECs), using liposome transfectionstaining method,Then constructed the breeding true cell strain for drug screening. As these three kinds of transfected cells containing the their own target gene enhancer, GLuc reporter gene and destroy rice blast hormone (Blastincidin) resistance selection gene, by detecting GLuc reporter gene expression level may reflect the activation status of various cell lines of NFκB, ERK, or JNK.With the depressant effect intensity of the drugs to activated NFκB, ERK and JNK to conclude the activity of the drugs.In addition, in this experimental system introduced Total the activator lipopolysaccharide (liPoPolysaccharide, LPS), owned by known NF-kappa B, ERK, JNK, results show that LPS timely and dosely dependent manner promote pBGLuc-of NFκB pBGLuc-ERK,plasmid pBGLuc-JNK transfected HUVECs cells Gaussia luciferase reporter gene expression, while pBGLuc transfected control cells was not obvious (enhancer motif is obviously for the purpose of gene action), it is demonstrated that pBGLuc-NFκB, pBGLuc-ERK, pBGLuc-JNK recombinant plasmid could express the purpose gene normally. Because LPS make the NF-kappa B, ERK and JNK activate to a certain level, in this way, it improved the basic level of luciferase, Gaussia luciferase in the experimental system, which was more conducive to the discovery of the inhibitory effect of test compounds, these three signaling pathways. The filter system has characteristics of stable, specific, high-throughput, using the system, we had10kinds of traditional Chinese medicine monomer to do activity screening, and found strong inhibitorycompounds to LPS-induced NF-kappa B, ERK, and JNK activated signaling pathway.Peimine, Ligustrazine,CB-301, CB-202and Peimine had significant inhibitory effect on JNK activation of LPS-induced.While In the ERK triggered pathway of LPS-induced activation, to which CB-301, Peininine, Calycosin glycosides and beta-sitosterol, etc. had significantly inhibited. At the signal pathway by NFκB induced, β-sitosterol, CB-202, CB-301and Calycosin glycoside have significant inhibition.Therefore, the study constructed the characteristic model of the high-throughput screening of anti-atherosclerosis suppressing the inflammation activity of drugs. By this high-flux screening model screened active drug to restrain the inflammatory AS, and because its mechanism is clear, so the model established the foundation for early drug research to further research in vivo activity of antiatherogenesis drug screening.