The Mechanism Of The Water Extract Of Danggui Buxue Decoction On The Immune Damage Of Bone Marrow Hematopoietic Stem Cells

Objective Explore the effective components of Danggui Buxue Tang(DBT)regulatory T cell immune response factor expression,restore the dynamic balance of T cell immune response network system and promote the proliferation and differentiation of HSCs,action mechanism of apoptosis inhibition of bone marrow cells,for the traditional Chinese medicine theory of qi and blood in the clinical application of traditional Chinese medicine treatment of autoimmune disease in the application of experimental basis and obj ective basisMethods Bone marrow cells were washed from the BALB/c mice limbs,and IFN-y intervention was used to make the bone marrow cell inflammatory injury model.The changes of the number and ratio of T helper(Th)cells in bone marrow cells were detected by flow cytometry to investigate the effect of the effective components of DBT on lymphocyte proliferation and differentiation.The changes of apoptosis rate of bone marrow cells and the number of HSCs in bone marrow cells were detected by flow cytometry to explore the protective effect of the effective components of DBT for enriching blood on cell damage caused by immune attack.The expression and distribution of T-bet,SLAM and SAP in bone marrow cells and the change of the combination of SAP and SLAM were detected by immunofluorescence,and the mechanism of the effective components of DBT to reduce the expression of IFN-γ was discussed.Western blot was used to detect the changes of bone marrow cells regulate the key factor of IFN-y expression,Fas/FasL key molecular signaling pathways and Jak/Stat signaling pathways and PRK/eIF2 cytokines gene and protein expression.To study the mechanism of DBT in regulating the apoptosis of HSCs cells,inhibiting the expression of key signaling factors such as gene transcription and cell division cycle,and promoting their proliferation and differentiationResults The flow detection results showed that after the intervention of the effective components of DBT,the ratio of CD3+CD4+ cells in the model group was not significantly changed compared with that in the normal group.Compared with the model group,CD3+CD4+ cells in the high-concentration group were significantly increased after 24h of drug intervention(P<0.01).Compared with the normal group,the ratio of CD3+CD8+ cells in the model group was significantly increased(P<0.01).The drug group was significantly lower than the model group(P<0.01).Compared with the normal group,the ratio of Thl cells in the model group was significantly increased(P<0.01).When the effective components of DBT can significantly reduce the ratio of Thl cells,the effect of the high concentration group is more obvious(P<0.01).Th2 cell ratio in the model group was significantly lower than that in the normal group(P<0.01).After 12h of drug intervention,the high concentration of Danggui and the combination of two drugs could be significantly increased(P<0.01).After 24h of drug intervention,the low concentration of Huangqi and Danggui can significantly increase the ratio of Th2 cells(P<0.01),while there was no significant difference between the other groups and the model group(P>0.05).The ratio of Th17 cells in the model group was significantly higher than that in the normal group(P<0.01).Compared with model group,each drug group was significantly reduced(P<0.01).The longer the intervention time is,the more obvious the effect.The effect is more obvious at low concentration.The ratio of Treg cells in the model group was significantly lower than that in the normal group(P<0.01).After 12h of drug intervention,the ratio of Treg cells was significantly increased in the combination group compared with the model group at low concentrations(P<0.01).Other groups showed no significant difference(P>0.05).After 24h of drug intervention,the drug groups were significantly lower than the model group(P<0.01).Compared with the normal group,the apoptosis rate of bone marrow cells in the model group was significantly increased(P<0.01).Compared with the model group,each drug group can significantly reduce,and the effect of low-concentration intervention is more obvious after 12h(P<0.01).The HSC quantitative in model group was significantly lower than that in the normal group(P<0.01).The effective components of DBT can significantly increase the number of HSCs(P<0.01).Fluorescence results showed that the expression of T-bet in the model group was significantly higher than that in the normal group(P<0.01).The effective components of DBT can significantly reduce it(P<0.01 or P<0.05).The combination ratio of SAP and SLAM was significantly decreased in the model group and the normal group(P<0.01).After 12h of drug intervention,the ratio of SAP to SLAM was significantly increased in the model group(P<0.01 or P<0.05).After 24h of intervention,there was no significant difference between the drug groups and the model group(P>0.05)Western Blot results showed that the expressions of Fas,Caspase-3,Caspase-9,eIF2a and p-eIF2a in the model group were significantly higher than those in the normal group(P<0.01),while those in the model group were significantly lower(P<0.01 or P<0.05)Compared with the normal group,the expression levels of stat-1,stat-3,p-stat-1 and p-stat-3 in the model group were increased(P<0.01),while those in the other drug groups were significantly decreased(P<0.01)Conclusion The active components of DBT can restore the dynamic balance of the T-cell immune response network and the proliferation and differentiation of HSCs in the bone marrow hematopoietic microenvironment by regulating the immune response network of T cells,effectively alleviate the immune-mediated damage of hematopoietic stem cells and bone marrow cells,and restore the balance of the hematopoietic microenvironment.