Effects of Jianpi Zishen Xiehuo Formula on macrophage polarization in patients with primary immune thrombocytopenia
Objective:To explore the possible mechanism of Jianpi Zishen Xiehuo Formula in the treatment of primary immune thrombocytopenia(ITP).Methods:Peripheral blood mononuclear cells(PBMC)were collected from ITP patients with spleen and kidney deficiency,fire injury to blood collateral syndrome and normal subjects.The cells were divided into normal group,model group,prednisone group and Jianpi Zishen Xiehuo Formula group.After the corresponding drug intervention for 48 h,the cells were collected and the expression of M1 and M2 macrophages was detected by flow cytometry.RT-PCR was used to detect NF-κB p65,IL-6,IL-10,TNF-α,TGF-β mRNA expression.Western Blot was used to detect NF-κB p65 and p-NF-κB p65,iNOS,Arg-1 protein expression.Results:Compared with the normal group,the expression of CD86 and iNOS protein in PBMC in the model group increased(P＜0.01),while the expression of CD206 and Arg-1 protein decreased(P＜0.01).The mRNA expression of IL-6,TNF-α increased(P＜0.01),the mRNA expression of IL-10,TGF-β decreased(P＜0.01).The expression of NF-κB p65 mRNA and NF-κB p65,p-NF-κB p65 protein increased(P＜0.05,P＜0.01).Compared with the model group,the expression of CD86 and iNOS protein in prednisone group and Jianpi Zishen Xiehuo Formula group decreased(P＜0.01,P＜0.05),while the expression of CD206 and Arg-1 protein increased(P＜0.01,P＜0.05),the mRNA expression of IL-6 decreased(P＜0.01),the mRNA expression of IL-10 increased(P＜0.05),and the mRNA expression of TNF-α decreased in the Jianpi Zishen Xiehuo recipe group(P＜0.05).The mRNA expression of NF-κB p65 and the protein expression of NF-κB p65 and p-NF-κB p65 decreased significantly(P＜0.05,P＜0.01).Conclusion:Jianpi Zishen Xiehuo Formula may reduce the polarization of Ml macrophages and improve the polarization of M2 macrophages by inhibiting NF-κB signal pathway in PBMC of ITP patients,then exert anti-inflammatory and immunosuppressive effects to treat ITP.
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