Expression Of Toll-like Receptor 4 In Alveolar Macrophages Of Diabetic Rats

ObjectiveToll-like receptors (TLRs) are a group of transmembrane proteins responsible for recognizing conserved pathogen-associated molecular patterns (PAMPs). TLRs play an bridge role in the activation of innate and adaptive immunity in vertebrate animals. In view of crucial significance in immunology, molecular biology and clinical study, TLRs have been much concerned since discovery. The role of TLRs signal transduction pathway on the onset and development of diabetes and its complications becomes a hot research. Experiments confirmed that TLR2 and TLR4 expression and signaling are increased in T1DM. Alveolar macrophages play an important role in the clearance of inhaled antigen and all aspects of pathogenic micro-organisms, my experiment investigate the altered expression of TLR4 in alveolar macrophages of diabetic rats after 4 weeks and 8 weeks.Materials and methods1.24 Male healthy wistar rats were divided into 3 groupsGroup A:the control group; group B:the 4 weeks-diabetic group; group C:the 8 weeks-diabetic group2. Capture and culture of alveolar macrophagesThe small fraction of cells after BAL and centificated were resuspended in RPMI-1640 meduium supplemented with 10% calf serum,the cells were plated into a 60mm petri dish with small glass slides placed in the dish, then were incubated for 2 h at 37℃in 5% CO2 with saturated humidity. Make macrophages adherent and dispose the non-adherent cells.3. Immmunocytochemical detection of TLR4 TLR4 was observed in every groups referencing to two step immmunocyto-chemica kids.4. Statistical analysisThe data were analyzed with SPSS13.0 software. The results were represented as Mean±SD, and comparision between groups were performed using analysis of variance, P value less than 0.05 was considered to denote significant difference.ResultsImmunocytochemical analysis for TLR4TLR4 of alveolar macrophages was expressed in the 4 groups respectively, but their strengths are different, groupB was higher than group A(P<0.05); groupC was significantly higher than groupB and groupA (P<0.05).Conclusion1,The expression of TLR4 of diabetic rats was higher than that of normal rats.2,When diabetes develop, the expression of TLR4 become much higher, which is indicated that TLR4 play an important roll on immunologic defence of alveolar macrophages of diabetic rats.