1.Biocompatibility of macrophages with quantum dots
Chong LI ; Cheng YAN ; Huimin JIN ; Shuiyun WU ; Yetao QIANG ; Nannan YAN ; Tengfei XIAO ; Sheng XIA
Chinese Journal of Tissue Engineering Research 2017;21(26):4217-4221
BACKGROUND: Compared with the traditional organic fluorescent dyes, quantum dots present good biomarker characteristics. Especially, quantum dots for cell labeling and targeted bioimaging present unique optical properties.OBJECTIVE: To investigate the biocompatibility of CdSe/ZnS quantum dots with mononuclear macrophages.METHODS: The macrophages RAW264.7 were inoculated into 96-well plates containing 0, 50, 100 mg/L CdSe/ZnS quantum dots for 1 or 2 hours. Then, the fluorescent signal was detected by flow cytometry. After 0-24 hours of culture,the fluorescence signal intensity of the macrophages cultured with 50 mg/L CdSe/ZnS quantum dots was detected by flow cytometry. After 18 hours of culture, quantitative PCR was used to detect the levels of tumor necrosis factor α and interleukin 1β in macrophages, and macrophage proliferation cell apoptosis were detected by MTT and flow cytometry,respectively.RESULTS AND CONCLUSION: The fluorescence signal intensity was positively correlated with the mass concentration of CdSe/ZnS quantum dots, and the intensity of the fluorescent signal was increased with the labeling time. After labeling using 50 mg/L CdSe/ZnS quantum dots, the fluorescence signal of macrophages increased continuously with time, and reached the peak at 18 hours. Compared with 0 mg/L quantum dot group, 50, 100mg/L quantum dot groups could significantly promote the expression of tumor necrosis factor α and interleukin 1β in macrophages (P < 0.01 or P < 0.05).The level of tumor necrosis factor α in the 100 mg/L quantum dot group was higher than that in the 50 mg/L quantum dot group (P < 0.01). The expression of interleukin-1β showed no difference between 50 and 100 mg/L quantum dot groups.The cell proliferation in the 50 and 100 mg/L CdSe/ZnS quantum dot groups was significantly higher than that in the 0 mg/L quantum dot group (P < 0.05), but there was no difference between the former two groups. In addition, 50 and 100 mg/L CdSe/ZnS quantum dots had no significant effect on apoptosis of macrophages. To conclude, CdSe/ZnS quantum dots could activate macrophages and promote their proliferation and secretion of inflammatory factors, but did not affect their apoptosis.
2.Immunomodulatory effects of butyrate on bone marrow derived dendritic cells
Yetao QIANG ; Shuiyun WU ; Mutian HAN ; Lu CHENG ; Huimin JIN ; Cheng YAN ; Chong LI ; Xia LIU ; Miaomiao ZHANG ; Qixiang SHAO ; Sheng XIA
Chinese Journal of Immunology 2015;(10):1315-1319
Objective:To investigate the effect of butyrate produced by bacterial metabolism on immune features of murine bone marrow-derived dendritic cells(BMDCs) and its potential mechanisms.Methods: BMDCs were prepared from bone marrow cells of C57BL/6 mice by being cultured with GM-CSF and IL-4.The expression of CD80,CD86,B7-DC and MHCⅡ on BMDCs and its pri-ming ability on the proliferation of OVA257-264 antigen specific CD8+T cell were analyzed by using Flow cytometry.The mRNA levels of IL-6 and IL-12 in BMDCs were detected by real-time fluorescence quantitative PCR(q-PCR).Simultaneously,Griess reaction and Western blot was used for analyzing the levels of NO2-in BMDCs culture supernatant and the ERK phosphortylation in BMDCs respectivly.Results:Butyrate could decrease the levels of CD80,CD86,MHCⅡand B7-DC,and downregulate the capability of BMDCs in priming the proliferation of CD8+T cells.Furthermore,the secretions of IL-6,IL-12,NO2-and the phosphorylation of ERK were sup-pressed.Conclusion:Butyrate down-regulats the immune functions of BMDCs via inhibition of ERK phosphorylation in TLR 4 signaling pathway.
3.Iron overload regulates atherosclerotic activity of foam cells induced by oxLDL
Xiaoyan WANG ; Xiaoyi ZOU ; Xiang ZHU ; Ting WANG ; Yetao QIANG ; Siyuan ZHOU ; Peng ZHANG ; Ping ZHANG
The Journal of Practical Medicine 2024;40(3):295-301
Objective To explore the roles of iron overload in pro-atherogenic activation of foam cells.Methods RAW264.7 and MOVAS cells were stimulated by oxLDL,ferrimine citrate and deferoxamine respectively.Prussian Blue and Oil Red O staining were used to detect iron deposition and foam cell.CCK-8 test,DHE probe,ELISA,RT-qPCR were performed to detect the cell death rate,reactive oxygen species(ROS)generation,lipid peroxidation molecules[glutathione peroxidase(GSH),glutathione peroxidase 4(GPX4),malondialdehyde(MDA)content]and the mRNA level of ATP binding cassette transporter A1(ABCA1),ATP binding cassette transporter G1(ABCG1),inductible nitris oxide synthase(iNOS),arginase-1(Arg-1),α-smooth muscle actin(α-SMA),smooth muscle 22 alpha(SM22a),osteopontin(OPN),Interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α).Results Iron overload could reduced reverse cholesterol transporters(ABCA1 and ABCG1),promote foam cells generation,increased cell death rate,induced the expression of lipid peroxidation molecules(GSH,GPX4,MDA),and promoted pro-inflammatory M1 marker of macrophage and synthetic marker expression of vascular smooth muscle cell(VSMC)and inflammatory cytokines(IL-1β,TNF-α).Conclusion Iron overload promotes the generation of foam cells derived from macrophages and smooth muscle cells and transform them into pro-atherosclerotic phenotype,aggravates cell lipid peroxidation and inflammatory reaction,which contributes to the progress of atherosclerosis.