A Study Of Sirna On Inhibiting The Expression Of Dendritic Celt CD40

Dendritic cells CD40is type I transmembrane phosphorylated glycoprotein withrelative molecular mass of4800~50000, the gene is located on20q11-20q13-2.It is akind of secondary signaling molecules mainly expressed on B cells、monocytes、macrophages、 DCs and a few fiber cells involved in the cellular and humoralimmune responses. Its ligand CD40L is a kind of type II transmembrane transporterproteins with relative molecular mass of about39000.They both belong to the TNFreceptor superfamily member.These receptor-ligand interaction is an important signaltransduction pathways in immune cells. Their combination can induce a variety oflymphocyte activation and produce a variety of pro-inflammatory factors (such as:IL-1, IL-2, IFN-gamma and INF-alpha, etc.) to attract white blood cells inextravascular, causing local tissue damage and dysfunction. Studies have shown thatCD40and CD40L costimulation plays an important role in the start and amplificationof immune response.Which is closely related to the development of infection andimmune-related diseases. If we design a specific molecules to block the interaction ofCD40and CD40L to induce T cell immune tolerance and make B cells secrete fewerinflammatory factors to reduce and prevent disease is very important,especially whenthe inflammation is very serious. The animal model has confirmed that it is usefullwhen we give CD40L antibody to mice with autoimmune and inflammatory diseases,including arthritis, pneumonia, fibrosis, experimental allergic encephalomyelitis,lupus nephritis, atherosclerosis atherosclerosis, acute pancreatitis and the ovarianYandeng induced by chronic collagen; CD40/CD40L system play a great role in theoccurrence and development of inflammatory bowel disease (IBD) through theexperiment that CD40L antibody in hapten-induced colitis model and otherexperimental and infection colitis model.RNA interference (RNA-the mediatedinterference, RNAi) is one of the hotspots in the treatment of disease currently. Thebasic principle is to be specifically cut into a length of19~21nt small interfering RNA (small interfering RNA, siRNA), as a guide sequence, and guide its reactionwith single-stranded target mRNA binding together to form the RNA-inducedsilencing complex (RNA induced silencing complex, RNSC). RNAi technology isincreasingly in-depth, and received a good results in a very wide range of applicationsin the medical field, including anti-viral treatment of tumors and neurodegenerativediseases treatment. However, application of siRNA in the treatment of IBD byinterventing dendritic cell CD40expression is seldomly reported. This study constructconstruct the expression vector by the combination of pSilencer and CD40sense andantisense RNAi fragment designed,then amplificated by DH5α and proved to bepSilencer-P450-of CD40RNAi and transferred to SD rat. Evaluate the expression levelof rat DC surface CD40and CD40mRNA by protein immunization mark, PCRmethod, the results showed that protein expression of CD40in dendritic cells issignificantly lower in the intervention group, gray scale scanning showed that theinhibit rate is about60%and CD40mRNA expression reduced by57.21%which idstatistically significant difference (P <0.05). The results lay the foundation of RNAtherapeutic for IBD in animal experiments or clinical application.