Toll-like Receptors 4 And Other Related Signal Molecules On The Synthesis And Secretion Function And Cells Proliferation Of Asthma Human Airway Smooth Muscle

Objective Bronchial asthma (asthma) is a kind of chronic airway diseases involved a variety of cytokines. The characteristic pathological changes of asthma are airway remodeling and airway inflammation.Airway smooth muscle cells (Human airway smooth muscle cells, HASMCs) proliferation is an important factor in airway remodeling. Recent studies found that, HASMCs not only proliferates, but also secrete cytokines and inflammatory mediators when co-stimulated with other inflammatory mediators, as the immune cells involved in the pathogenesis of asthma.We can more directly observed that HASMCs features and micro - environment changes in the similar asthma condition by building up the passively sensitized asthma model.wo evaluate function .By detecting the HASMCs proliferation and secretion of cytokines,we could evaluate the condition of HASMCs more scientifically and comprehensively in a state of asthma to explore that TLR4-related signaling molecules how to affect HASMCs proliferation and cytokine synthesis function. Also, attempt to explore that traditional medicine-Artemisinin and dexamethasone how to affect HASMCs TLR4 andTGF-β1 expression and cell proliferation in order to explore new ideas for treatment of asthma medicine.Methods By using Immunocytochemistry, ELISA, and MTT and other bio-molecular techniques, previously inflammatory cytokine TNF-α, PI3K inhibitor Ottawa Man penicillin (Wortmannin) and NF-κB inhibitor pyrrolidine dithio-amino - alkyl carboxylic acid (PDTC) as a tool for medicine to research on Cultured HASMCs , to explore TLR4 and other Signaling molecules how to affect cells proliferation and the function of synthesis and secretion about TGF-β1.Results 1.Asthma group compared with the normal HASMCs, the cell proliferation significantly increased (P<0.01), total protein synthesis was significantly higher (P<0.01), IgE secretion was significantly higher (P <0.01),Successfully building up the passively sensitized asthmatic model.2.Asthma group compared with the normal control group, secretion of TGF-β1 was significantly higher (P<0.01).3.TNF-αgroup compared with the normal control group,the number of positive expression of TLR4 was significantly higher (P<0.01), the cell proliferative response becomes significantly stronger(P <0.01), the concentration of TGF-β1 were significantly higher (P <0.01).4.TLR4 antibody group compared with the normal control group, the number of positive expression of TLR4 was significantly higher (P<0.01), the cell proliferative response becomes significantly weaker(P <0.01), the concentration of TGF-β1 were significantly lower (P <0.01).5.TNF-α+ Wortmannin group and TNF-α+PDTC group compared with the normal control group, the cell proliferative response becomes significantly weaker(P<0.05), the concentration of TGF-β1 were significantly lower (P <0.01).6.TNF-α+artemisinin group, TNF-α+dexamethasone group compared with the normal control group, the number of positive expression of TLR4 significantly reduced (P <0.01), the cell proliferative response decreased (P<0.05), the concentration of TGF-β1 were significantly lower (P <0.01). Conclusions 1.Passively sensitized HASMCs has stronger proliferation ability, a large number of synthesis and secretion of IgE, leading to micro-environment, changes might be the one ofthe important factors in airway remodeling.2.HASMCs proliferation is a prominent featureof in airway remodeling . TNF-αcan activate TLR4 and increase their expression on HASMCs, thereby enhancing HASMCs proliferation and secretion of TGF-β1 synthesis.HASMCs excessive proliferation and the release of a large number of TGF-β1 may be one of the pathological foundations which lead to airway remodeling in asthma.3.TLR4 antibody could inhibit the proliferation of asthmatic HASMCs and synthesis and secretion of TGF-β1 function, which may have a reverse or delay role on airway remodeling in asthma .4.TLR4 and PI3K, NF-κB have the ability that induce the HASMCs to secret TGF-β1.Inhibiting PI3K and NF-κB at the same time that it could significantly inhibit the cell proliferation and reduce the secretion of inflammatory factors.we can see that TLR4 related signaling molecules activated may be one of the mechanisms of pathogenesisgas leading to asthma airway remodeling.5.Artemisinin and dexamethasone can inhibit the cell proliferation and the synthesis and secretion of TGF-β1 of HASMCs by lowering the TLR4 expression on HASMCs ,which has a certain therapeutic effect to asthmatic airway remodeling.