ABSTRACT
Objective To observe the role of Niaogan Prescription against uropathogenic Escheriehia coli (UPEC) in invading bladder epithelial cells, and to discuss its mechanism of action. Methods After HTB-9 infected with UPEC, the cell invasion model was used to observe the protective effects of drug urine in human bladder cancer cell 5637 (HTB-9) against UPEC invasion. Effects of Niaogan Prescription on the main steps of signaling pathway: Toll-like receptor 4 (TLR4), adenylate cyclase 3 (AC3), cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), and Rac-1 was investigated by molecular biology techniques. Results Compared with the blank urine high dose group (10% volume fraction of blank urine), treatment with the drug-containing urine high dose group of Niaogan Prescription (10% volume fraction of drug urine) resulted in a significant decrease in UPEC invasion, the bacterial invasion rate of the rat blank urine group had no significant difference compared with the model group. Compared with the blank urine high dose group, the drug-containing urine high dose group of Niaogan Prescription improved TLR4 and AC3 protein expression, increased intracellular cAMP content, promoted PKA activation, and inhibited Rac1 activity. Conclusion Niaogan Prescription have the certain capacity against UPEC invasion of bladder epithelial cells, which mechanism is related to TLR4/cAMP signaling pathway. Niaogan Prescription; uropathogenic Escheriehia coli; invasion; human bladder epithelial cells; TLR4/cAMP signal pathway
ABSTRACT
AIM To observe the effects of Niaogan Decoction (Portulacae Herba,Taraxaci Herba) on expulsion of uropathogenic Escheriehia coli (UPEC) from human bladder epithelial cells and to explore its mechanism.METHODS After HTB-9 was infected with UPEC,drug urine exocytosed the UPEC invaded into urinary bladder epithelial cell were observed.With molecular biology technology,the mechanism of key factors to TLR4/cAMP signal pathway such as Toll-like receptor 4 (TLR4),adeuylate cyclase 3 (AC3),cyclic adenosine monophosphate (cAMP),protien kinase A (PKA),Myosin Ⅶ A and Rab interacting protein (MyRIP),Rab27b and Caveolin-1 were analyzed.RESULTS Treatment with Niaogan Decoction resulted in a significant increase in UPEC exocytosis compared with the blank control high dose group.Niaogan Decoction group improved TLR4,AC3,MyRIP,Rab27b and Caveolin-1 protein expression,increased intracellular cAMP content and promoted PKA activation.CONCLUSION Niaogan Decoction has the capacity to exocytose UPEC,its mechanism may relate to TLR 4/cAMP signal pathway.