Effects Of Serum Containing Astragalus Membranaceus And Hedyotis Diffusa On TGF - β <1> And FN Expression In Rat Glomerular Mesangial Cells

Purpose: By observing astragalus membranaceus and hedyotic diffusa effect onthe secretion of fibronectin and transformation growth factor-β1,toinvestigate the role of the target by astragalus membranaceus and hedyoticdiffusa of total flavonoids extract on glomerular disease.Material and method:（1）The angiotensin as a stimulating factor,ratglomerular mesangial cells in vitro,make its produce extracellular matrixaccumulation.The glomerular mesangial cells were incubated at37℃inhumidified5％CO2-95％air.The cell medium was left untouched for2-3days andchanged every other day until confluence.Glomerulus cell were grown to75％confluence,washed once with serum-free DMEM.According to the concentration of2x104cells/hole,MCs cells was seeded in24weii piates,each set of5hole.Cellswere made arrested in0.5％FBS DMEM for24h to synchronize the cell growth.After this period,the media was to DMEM.At last,the cells were collected tounder test.（2）twenty eight SD(specifi pathogen free),in which the half wasmale and the other half was female and180-220g body weight.The rats were randomlydivided into four group,namely:the control group,the astragalus membranaceusand hedyotic diffusa low does group,middle dose group and high dose group.N=7.Thenormal group,model group rats were given equal volime of distilled water,therest of the rats treated with corresponging drugs orally,1tims a day,for6days.2hours after the last administration,blood was taken from the abdominalaorta.Blood using a centrifuge,condition is1500rpm centrifugation for5minutre,taking supernatant.After56℃water bath,30min inactivation.By filtersterilization of0.22μm,-20℃preservation reserve.(3) By real-timefluorescent quantitive PCR method to detect mesangial cells cultured invitro,induced excessive secretion of fibronectin and transformation growth factor-β(4)By using enzyme–linked immmunosorbent(ELISA) to detect themesangial cells cultured in vitro,induced excessive secretion of fibronectin.Results:1. Comparison of measured, effect of rat glomerular mesangial cells and thesecretion of TGF-β1and FN by RT-PCR method showed, by angiotensin IIstimulation, secretion of TGF-β1and FN increased significantly, TGF-β1(5.3238±0.5938) and FN (7.6911±0.7689)and normalgroup(TGFβ1.0120±0.3288,FN1.0892±0.3475) showed significant difference (P <0.01). Radix astragali,Herba Hedyotis diffusae low dose, middle dose, high dose group (TGF-β1lowdose:0.9045±0.3118, middle dose:3.3949±0.4997， high dose:2.7856±0.6314; FNlow dose:：1.4647±0.0849,middledose:3.8603±1.0704, high dose:3.1882±0.2760)significantly reduced, there are significant differences compared with themodel group(P <0.01)). Comparison of the effects of using ELISA method to detectmesangial cells secreteFN,andnormal group721.02±232.35andmodel group1722.98±464.46comparison, there is significant differencebetween(P <0.01), astragalus root, Oldenlandia low dose, middle dose, highdose group (low dose group:938.44±454.12, middle dos837.25±284.44, the highdose group:1206.30±511.06)11.06differences compared with the model group,the low, middle dose group compared with the model group, there is significantdifference between(P <0.01), high dose group compared with the model group,there are differences,(P <0.01).2. Comparison of the of the treatment group the glomerular mesangialcells of rats in the secretion of TGF-β1and FN with RT-PCR method, the TGF-β1: there is significant difference between Radix Astragali, Herba Hedyotisdiffusae in high dose group, and low dose group(P <0.01). Between the middle,high dose group has no difference （P>0.05）. Comparison between FN: astragalusroot, Oldenlandia diffusa each dose group, there was significant differencebetween high dose group, and low dose group (P <0.01) There is no differencebetween, in high dose group （P>0.05）. Comparison of the effects of secretion assays in each treatment group glomerular mesangial cells by ELISA method ofFN, Radix Astragali, Herba Hedyotis diffusae between each dose group had nodifference（P>0.05）.Conclusion:1. astragalus membranaceus and hedyotic diffusa can inhibit Ang Ⅱinducedglomerulat mesangial cell extracellular matrix-fibronectin and transformationgrowth factor-β1.Through inhibit excessive expression of extracellular matrixto delay the occurrence and development of renal disease.clear target，spreadinghedyotis herb medicine astragalus.2. Each therapy group glomerular mesangial cells and the secretion of TGF-β1and fibronectin and the model group were significantly different, but betweenthe treatment groups did not differ, indicating that Radix Astragali, HerbaHedyotis diffusae fibronectin expression no dose-response relationship ofglomerular mesangial cells and the secretion of TGF-β1and fibe...