Effects Of Detoxification Removing Stasis And Nourishing Yin Prescription No TLR9Signal Path Of Macrophage Of Small Mice

Objective:By studying the effect of detoxification removing stasis and nourishing yin prescription on abnormal activation of TLR9signal pathway of RAW264.7cells after ODN stimulation, to explore the role of detoxification removing stasis and nourishing yin prescription is through regulate TLR9signal pathway to reduce dosage of glucocorticoid and play a synergism and detoxification role in the treatment of SLE.Method:RAW264.7cells grown in6-well plates in vitro were randomly divided into4groups, namely (1) blank serum group:900μl high sugar DMEM nutrient solution+100μl blank serum;(2) ODN stimulus group:900μl high sugar DMEM nutrient solution+100μl blank serum+ODN (ODN when they joined the final concentration for0.5uM);(3) ODN+dexamethasone group:900μl high sugar DMEM nutrient solution+100μl blank serum+ODN (ODN when they joined the final concentration for0.5uM)+dexameth-asone (DXM when they joined the final concentration of40ng/ml);(4)ODN+medi-cated serum group:900μl do not contain FCS high sugar DMEM nutrient solution+ODN (ODN when they joined the final concentration for0.5uM)+100μl medicated serum. After24hours, the expressions of TLR9,MyD88,NF-кB,IFN-a and IFN-βmRNA were analyzed by RT-PCR;the expressions of TLR9and NF-кB protein were analyz by Western blot.Result:Aftert reatment for24hours, the expression of TLR9,MyD88,NF-кB,IFN-α and IFN-β mRNA in ODN stimulus group were statistically different as compared with the blank serum group, the value of ODN stimulus group of TLR9,MyD88,NF-кB,IFN-a,IFN-β have been increased, its differences with statistical significance(P<0.05); compared with ODN stimulus group, ODN+dexamethasone group TLR9value have a certain degree of decline,but the difference was not statistically significant(P>0.05),MyD88,NF-кB,IFN-a value were declined, its difference with statistical significance (P<0.05), IFN-β value was significantly lower than the IFN-β value of ODN stimulus group, its difference with statistical significance (P<0.01);ODN+medicated serum group TLR9,MyD88, NF-кB, IFN-a value were declined as compared with ODN stimulus group, its differences with statistical significance (P<0.05), IFN-β value have a certain degree of decline, but the difference was not statistically significant (P>0.05). expression of TLR9and NF-кB protein in ODN stimulus group statistically different as compared with the blank serum group, the value of ODN stimulus group of TLR9,NF-кB have been increased, its differences with statistical significance (P<0.01); compared with ODN stimulus group, ODN+dexamethasone group TLR9value had a certain degree of decline, but the difference was not statistically significant (P>0.05), the value of NF-кB was declined, its difference with statistical significance (P<0.01); the value of TLR9, NF-кB in ODN+medicated serum group have been declined as compared with ODN stimulus group, the difference is statistically significant (P<0.01).Conclusion:Firstly, detoxification removing stasis and nourishing yin prescription can inhibit the expression of TLR9, MyD88, NF-к B,IFN-α,IFN-β mRNA and TLR9, MyD88protien in abnormal activation of TLR9signal pathway after ODN stimulated RAW264.7cells.Secondly,detoxification removing stasis and nourishing yin prescription may be through regulating TLR9signal pathway to reduce the amount of glucocorticoid and play a synergism and detoxification role in the treatment of SLE.