Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Postoperative sleep-restriction (SR) increased myeloid-derived suppressor cells (MDSCs) expansion and decreased splenic CD8 + cells activity via inhibiting splenic trefoil factor 2 (TFF2). ( A ) The expression of TFF2 in the spleen of SR mice with or without dexmedetomidine (Dex) treatment was analysed by real-time PCR (RT-PCR and western blotting. ( B ) Spleen weight in each group after 7 days of SR. ( C ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in SR mice with or without Dex treatment. ( D ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from SR mice with or without Dex treatment. ( E ) The mRNA and protein expression of TFF2 in the spleen were analysed by RT-PCR and western blotting in SR mice with or without vagus nerve stimulation (VNS). ( F ) Spleen weight in SR mice with the treatment of recombinant human TFF2 protein (rTFF2) or PBS. ( G ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in SR mice with the treatment of rTFF2 or PBS. ( H ) ELISPOT assay of the levels of IFN-γ and GrB in splenic CD8 + T cells from SR mice with the treatment of rTFF2 or PBS. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Derivative Assay, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Enzyme-linked Immunospot, Recombinant

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Subdiaphragmatic vagus nerve mediated the attenuated effects of dexmedetomidine (Dex) on sleep-restriction (SR)-induced decrease in splenic trefoil factor 2 (TFF2) expression, increase in the expansion of myeloid-derived suppressor cells (MDSCs) and decrease in CD8 + T cells activity. ( A ) The expression of TFF2 in the spleen of dexmedetomidine (Dex)-treated SR mice with or without bilateral sub-diaphragmatic vagotomy (SDV) was analysed by western blotting. ( B ) spleen weight in Dex-treated SR mice with or without SDV. ( C ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in Dex-treated SR mice with or without SDV. ( D ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from Dex-treated SR mice with or without SDV. All data represent mean ± SEM, n = 5; # P < 0.05, N.S., not significant.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Expressing, Derivative Assay, Activity Assay, Western Blot, Flow Cytometry, Enzyme-linked Immunospot

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Dexmedetomidine (Dex) attenuated postoperative sleep-restriction (SR)-induced decrease in splenic trefoil factor 2 (TFF2) expression, increase in myeloid-derived suppressor cells (MDSCs) expansion and decrease in splenic CD8 + cells activity via improving gut microbiota disturbance. ( A ) Western blotting analysis of splenic TFF2 expression in sham-operated mice and pseudo-germ-free mouse received fecal microbiota transplantation (FMT). ( B ) Spleen weight in sham-operated mice and pseudo-germ-free mouse received FMT. ( C , D ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in sham-operated mice and pseudo-germ-free mouse received FMT. ( E ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from sham-operated mice and pseudo-germ-free mouse received FMT. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Expressing, Derivative Assay, Activity Assay, Western Blot, Transplantation Assay, Flow Cytometry, Enzyme-linked Immunospot

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Subdiaphragmatic vagus nerve served as a bridge between gut microbiota and spleen after postoperative sleep-restriction (SR). ( A ) Western blotting analysis of splenic trefoil factor 2 (TFF2) expression in pseudo-germ-free mouse received fecal microbiota transplantation (FMT) with feces of SR mice, dexmedetomidine (Dex)-treated SR mice or Dex-treated SR mice with sub-diaphragmatic vagotomy (SDV). ( B ) Spleen weight in pseudo-germ-free mouse received FMT with feces of SR mice, Dex-treated SR mice or Dex-treated SR mice with SDV. ( C ) Flow cytometry analysis of spleen for CD11b + Gr-1 + myeloid-derived suppressor cells (MDSCs) in pseudo-germ-free mouse received FMT with feces of SR mice, Dex-treated SR mice or Dex-treated SR mice with SDV. ( D ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from pseudo-germ-free mouse received FMT with feces of SR mice, Dex-treated SR mice or Dex-treated SR mice with SDV. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Western Blot, Expressing, Transplantation Assay, Flow Cytometry, Derivative Assay, Enzyme-linked Immunospot

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Splenic trefoil factor 2 (TFF2) was essential to attenuate SR-induced reduced protective ability against Escherichia coli ( E. coli ) pneumonia, increased expression of IL-4 and IL-13 in the lung and M2 polarization of alveolar macrophages (AMs), and decreased phagocytic activity of AMs. ( A ) Enumeration of colony-forming units (CFU) per milliliter of bronchoalveolar lavage analyzed one day after E. coli pneumonia in postoperative SR mice treated with recombinant human TFF2 protein (rTFF2) or PBS. ( B ) ELISA determination of the concentrations of IL-4 and IL-13 in the lungs of postoperative SR mice treated with rTFF2 or PBS one day after E. coli pneumonia. ( C ) The phagocytic activity of alveolar macrophages from postoperative SR mice treated with rTFF2 or PBS. ( D ) Real-time PCR (RT-PCR) analysis of the mRNA expression of Arg1, YM and iNOS in alveolar macrophages from postoperative SR mice treated with rTFF2 or PBS. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Expressing, Activity Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Schematics illustrating the signaling mechanisms of sleep-restriction in postoperative immunosuppression and its treatment by dexmedetomidine. Sleep-restriction exaggerates intestinal flora disorder, furtherly decreases splenic trefoil factor 2 (TFF2) expression, which subsequently leads to the increase in myeloid-derived suppressor cells (MDSCs) numbers and decrease in splenic CD8 + T cells activity. Subdiaphragmatic vagus nerve (SVN) served as an important conduit of gut microbiota-spleen communication. SR-induced exaggeration of postoperative immunosuppression was characterized by increased expression of IL-4 and IL-13 in the lung, increased M2 polarization of alveolar macrophages (AMs), decreased phagocytic activity of AMs and thus decreased antimicrobial activity in E. coli pneumonia. Dexmedetomidine treatment during SR alleviated SR-induced decrease in postoperative immunosuppression through gut microbiota and SVN.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Expressing, Derivative Assay, Activity Assay

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: The sequences of primers for real-time PCR.

Article Snippet: Proteins (50 μg) were separated by electrophoresis on 10% SDS-PAGE gels (Beyotime Institute of Biotechnology, Shanghai, China), transblotted into PVDF membranes (Millipore; Merck KGaA), and incubated overnight at 4 °C with primary antibodies against TFF2 (1:500; Wuhan Boster Biological Technology, Ltd.) and β-actin (1:2,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: