The Relative Research Of Effects And Mechanisms Of The Method Of Benefiting Qi For (and) Relieving Superficies Syndrome On BD-2 In Controlling TLR4-NFκB Signal Transduction Pathways

ObjectiveTo explore the effects and mechanisms of the method of benefiting qi for (and) relieving superficies syndrome on BD-2 in controlling TLR4-NFκB signal transduction pathways between it and the method of relieving superficies syndrome.Methods1.Collect cardiac blood and produce drug-containing serum after using the method of intragastric administration to give Shensu drink and its parts liquid medicine to male SD rats.2.Subculture 16 human bronchial epithelial cells (16HBE).3.In order to build 16HBE inflammation model, the proliferation activity of 16HBE and the content of TNF-α, IL-8 in culture supernatant was determined at different time points (3h,6h,12h,24h) after LPS (lmg/L) stimulation.4.Pretreating 16HBE with PDTC for 30min, then, determining the content of hBD-2mRNA using RT-PCR at different time points (0.5h, lh,3h,5h,8h) after administration of drug-containing serum.5.Pretreating 16HBE with Anti-HumanCD284 for 1h, then, determining the content of NFκBp65mRNA and hBD-2mRNA using RT-PCR at different time points (0.5h, 1h,3h,5h,8h) after administration of drug-containing serum.Results1.Successfully cultured 16 human bronchial epithelial cells (16HBE).16HBE were polygonal, irregular and cobblestone-like monolayer growth.Generally, passaging one generation needed 3 to 5 days. After passaging for 3 ～ 4h, the cells started to adhere to bottom of the culture bottle, then, appeared split phase. The cells splitted to be clusters-like,network-like and finally connected into pieces.2.Succesfully builded 16HBE inflammation model. The proliferation activity of 16HBE and the content of TNF-α, IL-8 in culture supernatant significantly increased compared with those of blank group (p<0.05).3.The role of NFκB in Shensu Drink induing hBD-2mRNA expression in inflammatory 16HBE. After blocking NFκB with PDTC,the content of hBD-2mRNA in the group of full formula and group of nourishing qi to relive superficies significantly increased only at 1h compared with that of model group (p<0.05). The rest were no significant difference (P> 0.05).4.The role of TLR4 in Shensu Drink induing hBD-2mRNA expression in inflammatory 16HBE. After blocking TLR4 with Anti-HumanCD284,the content of NFicBp65mRNA and hBD-2mRNA in the group of full formula significantly increased only at 5h compared with that of model group (p<0.05), the content of NFKBp65mRNA and hBD-2mRNA in the group of nourishing qi to relive superficies significantly increased only at 3h compared with that of model group (p<0.05).The rest were no significant difference (P> 0.05).ConclusionsThe method of benefiting qi for (and) relieving superficies syndrome plays an important role on regulating TLR4-NF κB signaling pathway and highly expressing β-defensin-2;Shensu drink is a specific application of the method of relieving superficies syndrome in the upregulation of TLR4-NFκB pathway on high β3-defensin-2 expression;The method of benefiting qi for (and) relieving superficies syndrome can induce the body high expression of BD-2 to improve the ability of the body of the royal evil, and vice versa, the body high expression of BD-2 can also prompt the method of benefiting qi for (and) relieving superficies syndrome fully and strengthen the body immunity.