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Stimulator of interferon genes (STING) is a cytosolic DNA sensor which is regarded as a potential target for antitumor immunotherapy. However, clinical trials of STING agonists display limited anti-tumor effects and dose-dependent side-effects like inflammatory damage and cell toxicity. Here, we showed that tetrahedral DNA nanostructures (TDNs) actively enter macrophages to promote STING activation and M1 polarization in a size-dependent manner, and synergized with Mn2+ to enhance the expressions of IFN-β and iNOS, as well as the co-stimulatory molecules for antigen presentation. Moreover, to reduce the cytotoxicity of Mn2+, we constructed a TDN-MnO2 complex and found that it displayed a much higher efficacy than TDN plus Mn2+ to initiate macrophage activation and anti-tumor response both in vitro and in vivo. Together, our studies explored a novel immune activation effect of TDN in cancer therapy and its synergistic therapeutic outcomes with MnO2. These findings provide new therapeutic opportunities for cancer therapy.
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Aim To investigate the effect of Mahuang-Dahuang(MD)on inhibiting M1 polarization of alveolar macrophages, alleviating inflammatory injury and treating acute lung injury, and the underlying mechanism.Methods Abandoned rats were divided into normal control group(NC), model control group(MC), MD high-dose group(MD-H), medium-dose group(MD-M), low-dose group(MD-L), and DXMS group.Lipopolysaccharide(LPS)was injected intraperitoneally to prepare an acute lung injury(ALI)rat model.After the model was established, different doses of MD were administered by intragastric administration, and the pathological changes of lung tissues were observed; immunohistochemical method was used to detect the expression of alveolar macrophage marker F4/80,F4/80 and CD80, CD80 and IL-6 Co-expression; Flow cytometry was used to detect the content of F4/80 and CD80 in lung tissues; PCR was used to detect the relative expression of IL-6, INOS, TNF-α mRNA in lung tissues; Western blot was used to detect lung tissue CCR2, CCL2, p-p65, P65, p-p38MAPK, p38MAPK protein expression.Results Compared with NC group, rats in MC group had damaged alveolar structure, thickened pulmonary interstitial edema, and infiltrated a large number of inflammatory cells.Among them, the number of macrophages increased significantly, which promoted the increase in the expression of macrophages-related chemokines CCR2 and CCL2.The number and expression of M1 alveolar macrophages in lung tissues increased, the relative expression of M1 alveolar macrophages-related cytokines IL-6, INOS and TNF-α mRNA was up-regulated, and the target protein in the related pathways of alveolar macrophages M1 polarization p-NF-κBp65/NF-κBp65, p-P38MAPK/p38MAPK protein expression was up-regulated.Compared with MC group, the pathological status of lung tissues in MD high, medium, and low dose groups was significantly improved, the number of lung tissue M1 alveolar macrophages decreased, the relative expression of inflammatory factors IL-6, INOS and TNF-α mRNA was down-regulated, and the protein expression of CCR2, CCL2, p-NF-κBp65/ NF-κBp65 and p-P38MAPK/p38MAPK was reduced in lung tissues.Conclusions MD can inhibit the M1 polarization of alveolar macrophages, reduce the activation and release of inflammatory factors, inhibit inflammatory response, and prevent acute lung injury by regulating the expression of related indicators of NF-κB and MAPK signaling pathways.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST36) and "Feishu" (BL13) on M1 polarization of alveolar macrophages (AM) in rats with chronic obstructive pulmonary disease(COPD), so as to explore its anti-inflammatory mechanism underlying improvement of COPD. METHODS: Forty SD rats were randomly divided into normal and normal＋EA, COPD model and COPD＋EA groups (n＝10 in each group). The COPD model was established by simple fumigation. EA (4 Hz/20 Hz, 1 to 2 mA) was applied to bilateral ST36 and BL13 for 30 min, once every other day for 2 weeks. The pulmonary function including the forced vital capacity (FVC), forced expiratory volume in 0.1 and 0.3 s (FEV0.1, FEV0.3, FEV0.1/FVC, and FEV0.3/FVC) was detected by using a small animal respiratory function detector. Histopathological changes of the lung were displayed by H．E. staining. The contents of tumor necrosis factor-α (TNF-α) and induced nitric oxide synthase (iNOS) in the broncho alveolar lavage fluid (BALF) were assayed by ELISA. The expression of M1 polarization markers (CD86，iNOS), myeloid differentiation factor 88(MyD88) and nuclear factor-κB p65(NF-κB p65) in AM were detected by Western blot and quantitative real time-PCR, separately. The distribution and expression of CD86 in the lung were detected by immunohistochemistry. RESULTS: Following modeling, the levels of FVC, FEV0.1, FEV0.3, ratios of FVE0.1/FVC and FEV0.3/FVC were significantly decreased (P<0.01), while the contents of TNF-α and iNOS in the BALF, expression of CD86, iNOS, MyD88 and NF-κB p65 mRNAs and proteins in the AM, and CD86 immunoactivity in the lung were significantly increased in the model group relevant to the normal group (P<0.01). After the intervention, the decrease of the lung function and increase of the above-mentioned genes and proteins were all reversed in the COPD＋EA group (P<0.05, P<0.01). CONCLUSION: EA at ST36 and BL13 can reduce pulmonary inflammation in COPD rats, which may be related to its function in inhibiting M1 polarization of AM via down-regulating MyD88/NF-κB p65 signaling pathway.
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Objective To study if IRF1 could regulate the polarization by IRF 1 and M1 status and affect M1 media-ted antitumor function .Methods U937 derived M1 macrophage ( U937-M1 ) model was established .The cells were devided into 4 groups:the PMA pretreated unpolarized macrophage (M0), the PMA, IFN-γand LPS induced M1 macrophage (M1), the siRNA of IRF1 knocked down M1 macrophage (siIRF1) and the negative control siR-NA treated M1 macrophage (siC).Furthermore, the expression of CD86 and CD206 was detected by flow cytome-try, the M1/M2 associated markers (IL-12p35,IL-12p40,IL-23p19,IL-6,TNF-α/IL-10) and IFNB1 were ana-lyzed by qPCR,the expression of IL-12p70 and IL-10 was examined by ELISA, the expression of IRF1 and IRF5 was detected by Western blot , the proliferration and apoptosis of HCC were analyzed by CCK 8 and flow cytometry , respectively.Results Compared with the U937-M1, the IRF1 knocked down group showed impaired CD 86 expres-sion, but enhanced CD206 expreesion ( P<0.05 ); the expression of M1 related cytokines including IL-12p35, IL-12p40,IL-23p19,IL-6,TNF-αand IFNB1 was decreased, but M2 related cytokine IL-10 level was increased (P<0.01);the expression of IFN-β, IL-12p70 and IRF5 was impaired, but IL-10 was enhanced (P<0.05).In IRF1 knocked down U937-M1, the CCK8 analysis indicated that the M1 mediated anti-proliferation effects on hepatoma carcinoma cell were turned to pro-proliferation ( P<0.05);the flow cytometry showed that the M 1 mediated pro-ap-optosis effects were reversed to anti-apoptosis ( P<0.01 ) .Interestingly , IRF5 and IFN-βwere decreased at both mRNA and protein levels in IRF1 knocked down U937-M1 compared with the U937-M1 (P<0.01).Conclusions IRF1 may partly modulate IRF5 and IFN-β, and further regulate M1 polarization and its antitumor effects .
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Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with high fatality and morbidity and micro-glia/macrophage ( M/M) plays a vital role in SAH brain injury with complicated pathophysiological mechanism .This study was to ob-serve the effect of Nrf2 deficiency on M/M activation and M1 polarization after subarachnoid hemorrhage in mice . Meth ods We col-lected 70 wild-type ( WT) ICR mice and 35 Nrf2-knockout ( KO) mice to establish the SAH model by injecting fresh autologous blood into pre-chiasmatic cistern.WT mice were arranged into four groups: sham operation group, post operative day 1 (POD1) group, POD3 group and POD5 group.Then WT mice and Nrf2 Nrf2-knockout mice were divided into sham operation WT group , sham opera-tion KO group, SAH WT group and SAH KO group.Western blotting (WB) and immunohistochemistry (IHC) were applied to observe the activation and proliferation of M/M after SAH on WT mice .Difference in activation and M 1 polarization were observed by detecting Iba1 expression in WB and CD 16/32 +Iba1 +cells in immunofluorescence between WT and KO mice . Results Gray scale values of Iba1 expression by WB in WT mice are 0.491 ±0.039, 0.657 ± 0.069, 0.930 ±0.046 and 0.926 ±0.046;average optical intensity values of Iba1 expression by IHC in WT mice are 0.412 ±0.122, 0.625 ±0.135, 0.963 ±0.213 and 0.978 ±0.224.The data indica-ted that Iba1 expression increased in SAH KO group in comparison to SAH WT group on 1, 3, 5 day after SAH (P<0.05).Moreover, Nrf2 deficiency promoted the activation and polarization of M /M by increased Iba1 protein expression and CD16/32 +Iba1 +cells after SAH ( P<0.05). Conclusion SAH induces M/M activation and proliferation in mice, and Nrf2 deficiency promotes the activa-tion, proliferation and M1 polarization after SAH .