Journal: Stem Cell Research & Therapy

Article Title: Human umbilical cord mesenchymal stem cell-derived microvesicles alleviate pulmonary fibrosis by inhibiting monocyte‒macrophage migration through ERK1/2 signaling-mediated suppression of CCL2 expression

doi: 10.1186/s13287-025-04266-w

Figure Lengend Snippet: Effects of MSC-MVs on BLM-induced PF in mice. ( a ) Diagram of the experimental scheme. ( b ) HE staining of mouse lung tissue from the control, model, and MV groups. ( c ) Ashcroft score of three groups ( n = 5). ( d - e ) Masson staining of mouse lung tissues from the three groups of mice and the statistical analysis of collagen-volume fraction %( n = 5). ( f - k ) Immunohistochemical staining of collagen I, α-SMA and FN and comparative analysis of the average optical density (AOD) were performed to assess positive staining in lung tissue ( n = 5). ( l - o ) The corresponding levels of FN, collagen I and α-SMA in the lung tissues of mice in the control, model, and MV groups were determined by WB, with GAPDH as an internal reference The corresponding uncropped full-length blots are included in Supplementary Fig. d ( n = 4). The data are shown as the means ± SEMs. The Shapiro-Wilk test of the data > 0.05. One-way ANOVA followed by the Sidak multiple comparison test was used to identify differences among the three groups, and ∗ indicates the difference between the control and model groups or the difference between the model group and the MV group. ∗∗ p < 0.01. *** p < 0.001. Scale bar, 100 μm. Abbreviations: SEM: standard error of the mean

Article Snippet: Data with unequal sample sizes were analyzed via one-way ANOVA followed by the Sidak multiple comparison test (GraphPad Prism 8.0.1), whereas for non-normally distributed data, differences between two groups were determined via the Mann‒Whitney U test for unpaired observations.

Techniques: Staining, Control, Immunohistochemical staining, Comparison

Journal: Stem Cell Research & Therapy

Article Title: Human umbilical cord mesenchymal stem cell-derived microvesicles alleviate pulmonary fibrosis by inhibiting monocyte‒macrophage migration through ERK1/2 signaling-mediated suppression of CCL2 expression

doi: 10.1186/s13287-025-04266-w

Figure Lengend Snippet: MSC-MVs inhibited monocytes and macrophages in early BLM-induced PF in mice. ( a ) Flow cytometric analysis of neutrophils, monocytes and macrophages in the peripheral blood of animals in the control, model, and MV groups. ( b - d ) Statistical analysis of the flow cytometric results of neutrophils and macrophages in the peripheral blood of the three groups. ( e - g ) The serum levels of TNF-α, IL-6, and IL10 in the three groups were measured. ( h , i ) Flow cytometric analysis of neutrophils, monocytes, macrophages, M1 macrophages and M2 macrophages in the BALF of the three groups. ( j ) Statistical analysis of the number of total cells in the BALF of the three groups. ( k - o ) Statistical analysis of the flow cytometric results for neutrophils, monocytes, macrophages, M1 macrophages and M2 macrophages in the BALF of the three groups. ( p ) IF assay showing the protein expression of F4/80 and iNOS in the lung tissue of the three groups. Scale bar = 100 μm. ( q - s ) CBD was used to measure the levels of TNF-α, IL-6, and IL-10 in the lung tissue homogenates of the three groups. n = 5. The data are shown as the means ± SEMs. The Shapiro-Wilk test of the data > 0.05. One-way ANOVA followed by the Sidak multiple comparison test was used to identify differences among the three groups, and ∗ indicates the difference between the control and model groups or the difference between the model group and the MV group. ** p < 0.01. *** p < 0.001

Article Snippet: Data with unequal sample sizes were analyzed via one-way ANOVA followed by the Sidak multiple comparison test (GraphPad Prism 8.0.1), whereas for non-normally distributed data, differences between two groups were determined via the Mann‒Whitney U test for unpaired observations.

Techniques: Control, Expressing, Comparison

Journal: Stem Cell Research & Therapy

Article Title: Human umbilical cord mesenchymal stem cell-derived microvesicles alleviate pulmonary fibrosis by inhibiting monocyte‒macrophage migration through ERK1/2 signaling-mediated suppression of CCL2 expression

doi: 10.1186/s13287-025-04266-w

Figure Lengend Snippet: MSC-MVs inhibited chemotaxis through the CCL2/CCR2 axis to regulate the chemotaxis of monocytes and macrophages. ( a ) Bubble chart of the GO enrichment of 116 genes related to monocytes and macrophages. The color of the bubbles indicates the p value, and the size of the bubbles indicates the number of genes associated with the GO term. ( b ) Bubble chart of the GO enrichment of 28 genes related to the migration of monocytes and macrophages. The color of the bubbles indicates the p value, and the size of the bubbles indicates the number of genes associated with the GO term. ( c ) Flow cytometric analysis showing CCR2 expression in monocytes and macrophages in the peripheral blood of the control, model, and MV groups. ( d , e ) Statistical analysis of the flow cytometric results showing CCR2 expression in monocytes and macrophages in the peripheral blood of the three groups ( n = 5). ( f , h ) Representative images of immunohistochemical staining for CCR2 and CCL2. ( g , i ) Comparative analysis of the AOD values showing positive staining for CCR2 and CCL2 in lung tissues ( n = 5). ( j ) Serum CCL2 protein levels in the three groups( n = 5). ( k ) CCL2 levels in the lung homogenates of the three groups( n = 5). ( l ) Confocal microscopy revealed that MHS cells take up MSCs-MVs. MHS cells were stained with CFSE (green), and MSC-MVs were stained with PKH26 (red). ( m ) CCL2 mRNA levels in MHS cells in the control, LPS and LPS + MV groups ( n = 3). ( n ) The protein level of CCL2 in the MHS cell supernatants of the control, LPS and LPS + MV groups( n = 3). ( o ) Experimental program for assessing the migration of RAW264.7 cells. ( p ) Representative images of the wound-healing assay of RAW264.7 cells in the three groups. ( q ) Comparative analysis of relative migration rates (%) in the three groups( n = 3). The data are shown as the means ± SEMs. The Shapiro-Wilk test of the data > 0.05. One-way ANOVA followed by the Sidak multiple comparison test was used to identify differences among the three groups, and * indicates a difference between the control and model groups or between the model group and the MV group. ** p < 0.01. *** p < 0.001

Article Snippet: Data with unequal sample sizes were analyzed via one-way ANOVA followed by the Sidak multiple comparison test (GraphPad Prism 8.0.1), whereas for non-normally distributed data, differences between two groups were determined via the Mann‒Whitney U test for unpaired observations.

Techniques: Chemotaxis Assay, Migration, Expressing, Control, Immunohistochemical staining, Staining, Confocal Microscopy, Wound Healing Assay, Comparison

Journal: Stem Cell Research & Therapy

Article Title: Human umbilical cord mesenchymal stem cell-derived microvesicles alleviate pulmonary fibrosis by inhibiting monocyte‒macrophage migration through ERK1/2 signaling-mediated suppression of CCL2 expression

doi: 10.1186/s13287-025-04266-w

Figure Lengend Snippet: MSC-MVs induced ERK1/2 protein phosphorylation to inhibit CCL2 expression. ( a ) Bubble chart of the GO enrichment of 62 genes related to CCL2. The color of the bubbles indicates the p value, and the size of the bubbles indicates the number of genes associated with the GO term. ( b , d ) The corresponding levels of phosphorylated ERK1/2 (p-ERK1/2) and total ERK1/2 (t-ERK1/2) in the lung tissues of mice in the control, model, and MV groups were determined by WB, with GAPDH as an internal reference. The corresponding uncropped full-length blots are included in Supplementary Fig. b ( n = 4–5). ( c , e ) The corresponding levels of p-ERK1/2 and t-ERK1/2 in the MHS cells in the control, LPS and LPS + MV groups were determined by WB with GAPDH as an internal reference. The corresponding uncropped full-length blots are included in Supplementary Fig. c ( n = 4). ( f , g ) The mRNA and protein levels of CCL2 in MHS cells in the control, LPS, LPS + MV and LPS + LY3214996 groups ( n = 3). ( H ) Representative images of the wound-healing assay of RAW264.7 cells in the four groups. ( i ) Comparative analysis of relative migration rates (%) in the four groups ( n = 3). The data are shown as the means ± SEMs. The Shapiro-Wilk test of the data > 0.05. One-way ANOVA followed by the Sidak multiple comparison test was used to identify differences among the three groups, and * indicates the difference between the control and model groups or the difference between the model group and the MV group. ** p < 0.01. *** p < 0.001

Article Snippet: Data with unequal sample sizes were analyzed via one-way ANOVA followed by the Sidak multiple comparison test (GraphPad Prism 8.0.1), whereas for non-normally distributed data, differences between two groups were determined via the Mann‒Whitney U test for unpaired observations.

Techniques: Phospho-proteomics, Expressing, Control, Wound Healing Assay, Migration, Comparison