Journal: Theranostics

Article Title: Reversal of bleomycin-induced rat pulmonary fibrosis by a xenograft of human umbilical mesenchymal stem cells from Wharton's jelly

doi: 10.7150/thno.33741

Figure Lengend Snippet: Transplantation of HUMSCs reduced the activation of fibroblasts and promoted the expression of TLR-4 in the left lungs of rats with PF. (A) Left lung sections were labeled with anti-α-SMA antibody to indicate activated fibroblasts in the left lungs of each group. (B) Western blot images represent the contents of α-SMA in the rats' left lungs. Quantitative results of α-SMA from western blotting are shown. The results indicated that activated fibroblasts significantly increased from Day 7 to 49. The activated fibroblasts significantly decreased after the transplantation of high doses of HUMSCs. (C) Rat Acta 2 mRNA were extracted from left lung suspensions and analyzed by quantitative RT-PCR, demonstrating that a significant decrease in acta2 expression in the BLM+HUMSCs (HD) group on Day 49. (D) Cell number in bronchoalveolar lavage fluid (BALF) of the BLM+HUMSCs (HD) group reduced significantly compared with that in the BLM group on Day 49. (E) The immunohistostaining of anti-TLR-4 showed that some TLR-4 positive cells were localized in connective tissue (arrow heads), however, some in the alveoli (arrows). (F) To further explore whether TLR-4 expression was localized in M2 macrophage, the left lung tissue sections from the BLM+HUMSCs (HD) group on Day 49 were subjected to double-staining with anti-TLR-4 (green) and anti-CD163 (red) antibodies. (G) The contents of TLR-4 in the rats' left lungs were detected through western blotting. The quantitative results indicated that TLR-4 significantly increased following the transplantation of HUMSCs. (H) Quantitative analysis of real-time RT-PCR of rat TLR-4 mRNA in the left lung tissue lysate demonstrated that rat TLR-4 mRNA was increased in the BLM+HUMSCs (HD) group (n = 3/group). (I) The result of RT-PCR of human TLR-4 revealed that human TLR-4 was only found in cell lysate of A549 cell line, but not in the lungs of all groups. (J) Left lung samples from the Normal, BLM, and BLM+HUMSCs (HD) groups on Day 49 were subjected to Human Cytokine Antibody Array analysis. The results suggested that the transplantation of HUMSCs increased human FGF-6 and IGF-1 concentrations in rats with pulmonary fibrosis. (K) Analyses were conducted using Rat Cytokine Antibody Array. The results indicated that the transplantation of HUMSCs stimulated rats with pulmonary fibrosis to produce higher amounts of β-NGF, fractalkine, and GM-CSF in their left lungs. ✱ vs the Normal group, p < 0.05. # vs the BLM group, p < 0.05. ♠ vs the BLM+HUMSCs (LD) group, p < 0.05. ⧫ vs the BLM+HUMSCs (HD) group, p < 0.05.

Article Snippet: To elucidate which human cytokines released from HUMSCs were involved in the treatment of rat PF, a human protein cytokine kit (AAH-CYT-2000, RayBio Human Cytokine Antibody Array C Series 2000, RayBiotech) was used to screen the expression of 174 human cytokines (n = 3/group).

Techniques: Transplantation Assay, Activation Assay, Expressing, Labeling, Western Blot, Quantitative RT-PCR, Double Staining, Reverse Transcription Polymerase Chain Reaction, Ab Array

Journal: Molecules

Article Title: Polyphenol Effects on Splenic Cytokine Response in Post-Weaning Contactin 1-Overexpressing Transgenic Mice

doi: 10.3390/molecules24122205

Figure Lengend Snippet: Levels of IL-12 in splenic supernatants from cell cultures of postnatal day 8 pups. Abbreviations: wt = wild type; +/+ = homo, +/− = hetero; PMA: phorbol myristate acetate, POP: suckling pups that received milk from dams administered with polyphenols. Evaluation of cytokines was performed using a cytometric bead array (CBA) mouse soluble protein Flex Set kit, as described in the Materials and Methods section. Statistical analysis was performed using Bonferroni’s test for comparison between groups. GraphPad Prism statistical software release 5.0 for Windows Vista was used. Statistical significance was set at p < 0.05.

Article Snippet: Cell supernatants obtained from single mice were incubated with the cytometric bead array (CBA) mouse soluble protein Flex Set kit (Becton Dickinson, Milan, Italy).

Techniques: Comparison, Software