Objective·To study the regulatory effect of saikosaponin A(SSA)on the differentiation,apoptosis,and immunosuppressive function of myeloid-derived suppressor cells(MDSCs)in mice,and to explore their molecular mechanism.Methods·Recombinant mouse granulocyte-macrophage colony-stimulating factor(GM-CSF)was used to induce the differentiation of mouse bone marrow cells(BMCs)into MDSCs,or magnetic beads were used to sort MDSCs from tumor-bearing mice.After treating MDSCs with different concentrations(0,2.5,5.0 mg/L),flow cytometry(FCM)was used to detect the differentiation and apoptosis of MDSCs,as well as the expression levels of liver X receptor α(LXRα),arginase-1(Arg-1),and reactive oxygen species(ROS).At the same time,the effects of MDSCs on the proliferation function of T cells,and the effects on the nuclear factor κB(NF-κB),and signal transducer and activator of transcription 1(STAT1)signaling pathways were also detected.The mRNA levels of LXRα and Arg-1 were detected by quantitative real-time PCR(qPCR).Mice were given SSA by gavage(ig)or intraperitoneal injection(ip),and the mice were sacrificed after administration;and body mass,spleen weight,and spleen index were calculated.FCM was used to detect the proportion of immune cells in the spleen of mice.Results·SSA could up-regulate the expression level of LXRα in MDSCs,reduce the differentiation of M-MDSCs,induce apoptosis of MDSCs,reduce the expression levels of Arg-1 and ROS in MDSCs,and reduce the inhibitory effect of MDSCs on T cell proliferation.SSA inhibited the phosphorylation levels of NF-κB and STAT1 in MDSCs.The mice treated with SSA by gavage or intraperitoneal injection showed no significant changes in body weight and spleen index.Both modes of administration can reduce the proportion of MDSCs and their subset M-MDSCs in mice,but had different degrees of regulatory effects on other immune cells.Conclusion·SSA could regulate the differentiation and apoptosis of MDSCs,and inhibit their immunosuppressive function,which may be associated with the up-regulation of LXRα expression,and down-regulation of the NF-κB and STAT1 signaling pathways in MDSCs.

Objective·To study the regulatory effect of saikosaponin A(SSA)on the differentiation,apoptosis,and immunosuppressive function of myeloid-derived suppressor cells(MDSCs)in mice,and to explore their molecular mechanism.Methods·Recombinant mouse granulocyte-macrophage colony-stimulating factor(GM-CSF)was used to induce the differentiation of mouse bone marrow cells(BMCs)into MDSCs,or magnetic beads were used to sort MDSCs from tumor-bearing mice.After treating MDSCs with different concentrations(0,2.5,5.0 mg/L),flow cytometry(FCM)was used to detect the differentiation and apoptosis of MDSCs,as well as the expression levels of liver X receptor α(LXRα),arginase-1(Arg-1),and reactive oxygen species(ROS).At the same time,the effects of MDSCs on the proliferation function of T cells,and the effects on the nuclear factor κB(NF-κB),and signal transducer and activator of transcription 1(STAT1)signaling pathways were also detected.The mRNA levels of LXRα and Arg-1 were detected by quantitative real-time PCR(qPCR).Mice were given SSA by gavage(ig)or intraperitoneal injection(ip),and the mice were sacrificed after administration;and body mass,spleen weight,and spleen index were calculated.FCM was used to detect the proportion of immune cells in the spleen of mice.Results·SSA could up-regulate the expression level of LXRα in MDSCs,reduce the differentiation of M-MDSCs,induce apoptosis of MDSCs,reduce the expression levels of Arg-1 and ROS in MDSCs,and reduce the inhibitory effect of MDSCs on T cell proliferation.SSA inhibited the phosphorylation levels of NF-κB and STAT1 in MDSCs.The mice treated with SSA by gavage or intraperitoneal injection showed no significant changes in body weight and spleen index.Both modes of administration can reduce the proportion of MDSCs and their subset M-MDSCs in mice,but had different degrees of regulatory effects on other immune cells.Conclusion·SSA could regulate the differentiation and apoptosis of MDSCs,and inhibit their immunosuppressive function,which may be associated with the up-regulation of LXRα expression,and down-regulation of the NF-κB and STAT1 signaling pathways in MDSCs.

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous cell population of myeloid origin with immunosuppressive function. MDSCs can inhibit the normal immune response of the host to tumor by inhibiting the activity of T cells, natural killer cells, macrophage, B cells, and promoting the proliferation of Treg and other mechanisms. Therefore, MDSCs are potential targets for tumor immunotherapy. However, whether immunotherapy or other treatments, the effectiveness of monotherapy is limited, so immunotherapy combined with other treatments is a breakthrough combination. In recent years, immunotherapy combined with other treatments has shown initial results, with better results than monotherapy. It has become a research hotspot in tumor treatment and a new treatment method in the future. This article summarized the research progress on the effects of immunotherapy combined with chemotherapy, radiotherapy, Chinese herbal extract and Immunotherapy combination on MDSCs, in order to provide theoretical support for the clinical application of immunotherapy based combined tumor treatment measures.

Objective To investigate the effect of Angelica polysaccharide(APS)on the differentiation and function of M2 macrophages and underlying molecular mechanism.Methods Mouse bone marrow derived macrophages(BMDM)and M2 macrophages were induced and treated with APS(0,80,160,320 μg/mL);Mouse peritoneal macrophages were isolated and treated with APS(0,160 μg/mL).Flow cytometry(FCM)was used to detect mannose receptor(MR),CD11b,F4/80,CD163,and ARG-1 expression levels,apoptosis,and phagocytic ability of M2 macrophages and peritoneal macrophages.Mice were randomly divided into APS gavage group and control group,APS was intragastrically administered to mice,and macrophage MR expression level in blood and spleen were detected by FCM.Fluorescence microscopy was used to observe the morphology of BMDM-differentiated M2 macrophages.RT-qPCR was employed to detect the mRNA expression levels of MR and ARG-1 in M2 macrophages.Immunofluorescence assay was performed to detect the expression of the proteins related to molecular mechanism of differentiation and function of M2 macrophages.Results Compared with the 0 μg/mL APS group,the MR expression level in the M2 macrophages was decreased with the increase of APS concentration within a certain concentration range(80～320 μg/mL),and the MR expression level in peritoneal macrophages was also decreased in the 160 μg/mL APS treatment group(P＜0.01).The expression level of macrophage MR was also significantly decreased in peripheral blood and spleen in the APS gavage mice than the control group(P＜0.05).Compared with the 0 μg/mL APS group,the expression levels of CD11b,F4/80,and CD163 in the macrophages were increased in the 80～320 μg/mL APS treatment groups(P＜0.01).The morphology of macrophage had changed,from mostly spindle-shaped and pseudopodia to mostly round or irregular,and even a few cells with pseudopodia.APS induced apoptosis in M2 macrophages(P＜0.05).Compared with the 0 μg/mL APS group,M2 macrophages treated with 160 μg/mL APS had an increased ability to phagocytose fluorescent microspheres(P＜0.01),but the expression level of ARG-1 was decreased(P＜0.01).The mRNA expression of MR and ARG-1 in M2 macrophages was decreased(P＜0.05).The mean fluorescence intensity of phosphate acidified-signal transducers and activators of transcription 6(p-STAT6)-positive signals in M2 macrophages was significantly reduced in the 160 μg/mL APS-treated group(P＜0.05).Conclusion APS has bidirectional regulation on the differentiation and function of M2 macrophages,which may be associated with its downregulation of signal transducers and activators of transcription 6(STAT6)signaling pathway.

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.