Astragalus Polysaccharid On The Effect Of Adipose Tissue Macrophage Activation Mechanism

Background Insulin resistance (insulin resistance IR) is a common pathogenesis of many diseases, is an inflammatory condition. Macrophages in adipose tissue (adipokse tissue macrophages ATMs) has an important role in IR. Recent studies have shown, astragalus polysaccharides (Astragalus Polysaccharide APS) can improve the IR, but its mechanism is still not completely clear. Our previous studies have found that APS can reduce the number of ATMs while improving IR. While the increase in the number of ATMs will aggravate the adipose tissue of low-grade inflammation, inflammatory adipokines, IR. ATMs has become a new target in the treatment of IR. A review of the literature, there are few reports have the effect of APS on ATMs. APS reduce the ATMs effect is affected by fat cells? Or direct effects on macrophages? The mechanism by which reduced ATMs? These questions are still not clear, need further exploration.Objectives To investigate the effects and mechanism of APS on ATMs. Clear action site APS, is affected by adipocytes, or direct effects on macrophages; to observe the expression of inflammatory cytokines, to explore the intervention of APS on macrophage activation effect.Methods (1) Cell culture:adipocytes by 3T3-L1 cells, macrophages by ANA-1 cell line.The 3T3-L1 preadipocyte cells were seeded on culture plate, containing 10% fetal calf serum sugar DMEM culture at a temperature of 37℃,5%CO□training.The cells grow to full fusion to 80-90%,2 days later, and with final concentration of 0.5mmol/L IBMX, 1umol/L DEX and the final concentration of lOug/ml insulin 10% calf serum sugar DMEM culture 48h;48h changed after the final concentration of 10ug/ml containing 10% calf serum insulin, high glucose DMEM and 48h culture;After changing the 48h containing 10% calf serum to culture high glucose DMEM; After every 2 days to replace the high glucose DMEM containing 10% calf bovine serum medium once;Differentiation of 8 to 12 days,90% of the 3T3-L1 cells were mature adipocytes, for testing Oil Red O identification.(2) This research adopts the control design.The experiments were divided into three groups:The normal control group (NC),APS intervention group (AF) of adipocyte,Macrophage APS intervention group (AM),Each group should develop 8 multiple holes.The mature adipocytes of less than 90% out of the last group, each group choose 6 holes as control test. By Transwell co cultured with macrophages, will species in the indoor, adipose cell in indoor, the aperture of 0.4um film. Adipocyte differentiation and maturation of 0.100g/L after 3T3-L1, APS were added to the adipocyte culture fluid and macrophage medium as intervention factors, the control group cells were cultured without APS; 48 hours after removal chamber, removing the culture liquid, adding cell lysate, collecting cell lysate by semi quantitative reverse transcription PCR (RT-PCR) detected by nitric oxide synthase (iNOS) expression. At the end of the experiment using ELISA method (ELISA) to detect the expression of IL-6, TNF-a liquid cultured in cytokine.(3) Statistical analysis of the experimental results by using the software of SPSS 13.0, to x ± s said the findings. Using t test, test level using 0.05, P<0.05 had significant difference.Results (1)ELISA test results showed: the comparison of group IL-6:AF and group NC the room under the lower chamber, P< 0.05, the difference has statistical significance. Comparison of AM group and NC group on the upper chamber room, group AM and group NC on the upper chamber room, AF room and the room under the NC group, P> 0.05, the difference was not statistically significant. The TNF-a: comparison between AF group and NC group room under the lower chamber, P< 0.05, the difference has statistical significance. Comparison of AM group and NC group on the upper chamber room, group AM and group NC on the upper chamber room, AF room and the room under the NC group, P> 0.05, the difference was not statistically significant.(2)RT-PCR test results showed: Room iNOS NC group on the high expression of AF group, AM group on ventricular chamber and iNOS expression is decreased in AF group, but the expression on ventricular volume is lower, the difference was statistically significant (P<0.05).Conclusions 1.APS ATMs to reduce the action site is adipocytes.2.APS can inhibit the release of inflammatory factors in adipocytes, thereby reducing the activation of macrophages.