Anti-inflammative effect of glycyrrhizin on rat thermal injury via inhibition of high-mobility group box 1 protein.

AIM: Glycyrrhizin (Gly) has been reported as an inhibitor of extracellular HMGB1 (high-mobility group box 1 protein) cytokine's activity, and protects spinal cord, liver, heart and brain against ischemia-reperfusion-induced injury in rats. The purpose of this study was to investigate the protective effect of Gly in rat skin thermal injury model and to elucidate the underlying mechanisms. METHODS: Twenty-four male Sprague-Dawley rats (200-250g) were randomly divided into control group, vehicle-treated and Gly-treated burn groups, each group contained eight animals. In the latter two groups, rats were subjected to 30% TBSA (Total Body Surface Area) full-thickness scald injury. In Gly-treated burn group, glycyrrhizin (60mg/kg) was administered intraperitoneally immediately after and at 24th hour burn; in vehicle-treated burn group, Ringer's solution (4ml/kg, as a vehicle) was administered intraperitoneally immediately after and at 24th hour burn. The animals were sacrificed at 48h after injury. Aortic blood samples were obtained to detect tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) with ELISA (Enzyme-Linked Immuno Sorbent Assay) kits. Lung, liver and kidney tissue samples were collected to determine the expression of HMGB1 mRNA and protein. HMGB1 mRNA level was semiquantitatively measured by Real-Time PCR using ß-actin as an internal standard, and protein expression of HMGBI was determined by Western blot. RESULTS: Severe skin scald injury caused a significant increase in plasma TNF-α and IL-1ß versus the control group (P<0.001) in 48h after burns. Intraperitoneal administration of Gly (60mg/kg) significantly reduced the levels of serum TNF-α and IL-1ß (P<0.01). Gly treatment reduced these biochemical indices accompanied by lower level of HMGB1 protein (P<0.05) and mRNA expression (P<0.01). CONCLUSION: These results demonstrate that Gly possesses an anti-inflammation effect to protect the remote organs from burn-induced injury.

Inhibition of oxygen-glucose deprivation-induced apoptosis of human adipose-derived stem cells by genetic modification with antiapoptotic protein bcl-2.

BACKGROUND: Adipose-derived stem cells (ADSCs) have become a promising tool for a wide range of cell-based therapies. However, transplanted ADSCs do not survive well under ischemic conditions. In this study we aimed to inhibit oxygen-glucose deprivation (OGD)-induced apoptosis of human ADSCs by genetic modification with antiapoptotic protein Bcl-2. METHODS: After isolation and culture, the phenotypes of human ADSCs at passage 3 were analyzed by flow cytometry. Then, genetic modification of ADSCs with Bcl-2 was carried out. Bcl-2 gene transfection was verified by Western blot analysis and multipotent differentiation properties were evaluated in Bcl-2-modified ADSCs (Bcl-2-ADSCs). Apoptosis was evaluated by a TUNEL assay under ischemic conditions induced by OGD. Apoptotic nuclei were also assessed and quantified by Hoechst staining. RESULTS: The cultured ADSCs expressed stem cell-associated markers CD29, CD34, CD44, and CD90, but not fibroblast marker HLA-DR or hematopoietic stem cell marker CD133. The Bcl-2 gene was transferred into ADSCs efficiently, and Bcl-2-ADSCs differentiated into adipocytes, chondrocytes, and osteoblasts. In addition, Bcl-2 overexpression reduced the percentage of apoptotic Bcl-2-ADSCs by 38 % under OGD. CONCLUSION: Our results indicate that Bcl-2 overexpression through gene transfection inhibits apoptosis of ADSCs under ischemic conditions. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Acceleration of wound healing in acute full-thickness skin wounds using a collagen-binding peptide with an affinity for MSCs.

Mesenchymal stem cells (MSCs) have been accepted as a promising cell source in tissue repair and regeneration. However, the inability to enrich MSCs in target areas limits their wide application. As a result, it has been a major goal to induce MSCs to be abundantly and specifically recruited to the injury site. In this study, a peptide with a specific affinity for MSCs (E7 peptide) was immobilized to a collagen scaffold via a collagen-binding domain (CBD) to construct a functional collagen scaffold. In addition, the hypothesis that this method could recruit MSCs specifically was evaluated in a porcine model. In vivo investigations indicated that due to the immunore-action, the CBD-MSC-peptide collagen scaffold enhanced MSC adhesion and infiltration and promoted wound healing. At day 7 after surgery, we found more infiltrating cells and capillaries in the Collagen/CBD-E7 peptide group compared to the Scaffold group. At day 14, 21 and 28, a faster healing process was observed in the Collagen/CBD-E7 peptide group, with significant differences compared with the other groups (P < 0.05, P < 0.01). The results demonstrate the potential use of targeted therapy to rapidly heal skin wounds.