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Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts: involvement of C-Jun N-terminal kinase, cyclin D1, and NF-κB.

Wang W, Du Z, Yan J, Ma D, Shi M, Zhang M, Peng C, Li H - PLoS ONE (2014)

Bottom Line: MSCs were successfully induced into hepatocyte-like cells, osteoblasts and adipocytes in vitro.Increased activities of AP-1 and NF-κB, as well as elevated expressions of p-c-Jun, cyclin D1, and proliferating cell nuclear antigen (PCNA), were also found in MSCs therapy group.These data suggest that MSCs therapy promotes hepatocyte proliferation and prolongs survival in SFSLT by reducing ischemia reperfusion injury and acute inflammation, and sustaining early increased expressions of c-Jun N-terminal Kinase, Cyclin D1, and NF-κB.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China; Department of Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou University School of Medicine, Zhengzhou, Henan Province, China.

ABSTRACT

Background: The therapeutic potential of mesenchymal stem cells (MSCs) has been highlighted recently for treatment of acute or chronic liver injury, by possibly differentiating into hepatocyte-like cells, reducing inflammation, and enhancing tissue repair. Despite recent progress, exact mechanisms of action are not clearly elucidated. In this study, we attempted to explore whether and how MSCs protected hepatocytes and stimulated allograft regeneration in small-for-size liver transplantation (SFSLT).

Methods: SFSLT model was established with a 30% partial liver transplantation (30PLT) in rats. The differentiation potential and characteristics of bone marrow derived MSCs were explored in vitro. MSCs were infused transvenously immediately after graft implantation in therapy group. Expressions of apoptosis-, inflammatory-, anti-inflammatory-, and growth factor-related genes were measured by RT-PCR, activities of transcription factors AP-1 and NF-κB were analyzed by EMSA, and proliferative responses of the hepatic graft were evaluated by immunohistochemistry and western blot.

Results: MSCs were successfully induced into hepatocyte-like cells, osteoblasts and adipocytes in vitro. MSCs therapy could not only alleviate ischemia reperfusion injury and acute inflammation to promote liver regeneration, but also profoundly improve one week survival rate. It markedly up-regulated the mRNA expressions of HGF, Bcl-2, Bcl-XL, IL-6, IL-10, IP-10, and CXCR2, however, down-regulated TNF-α. Increased activities of AP-1 and NF-κB, as well as elevated expressions of p-c-Jun, cyclin D1, and proliferating cell nuclear antigen (PCNA), were also found in MSCs therapy group.

Conclusion: These data suggest that MSCs therapy promotes hepatocyte proliferation and prolongs survival in SFSLT by reducing ischemia reperfusion injury and acute inflammation, and sustaining early increased expressions of c-Jun N-terminal Kinase, Cyclin D1, and NF-κB.

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MSCs therapy kept the activity of transcription factors AP-1 and NF-κB, and increased the expressions of phosphorylated c-Jun (p-c-Jun) and Cyclin D1 (CyD1).A. Activation of NF-κB and AP-1 was evaluated by electrophoretic mobility shift assay at 1, 6 and 24 h after reperfusion. Compared with the 30PLT+PBS group, the activity of NF-κB and AP-1 was considerably enhanced at 24 h after reperfusion in the 30PLT+MSCs group; B. Expressions of p-c-Jun and CyD1 were detected by western blotting at 6 h, 24 h, 72 h, and 7 days after reperfusion. Expressions of p-c-Jun and CyD1 were remarkably increased at 72 h after reperfusion. (Mean ± SD; #p<0.05; *p<0.01; NS- nonsignificant.)
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pone-0112532-g007: MSCs therapy kept the activity of transcription factors AP-1 and NF-κB, and increased the expressions of phosphorylated c-Jun (p-c-Jun) and Cyclin D1 (CyD1).A. Activation of NF-κB and AP-1 was evaluated by electrophoretic mobility shift assay at 1, 6 and 24 h after reperfusion. Compared with the 30PLT+PBS group, the activity of NF-κB and AP-1 was considerably enhanced at 24 h after reperfusion in the 30PLT+MSCs group; B. Expressions of p-c-Jun and CyD1 were detected by western blotting at 6 h, 24 h, 72 h, and 7 days after reperfusion. Expressions of p-c-Jun and CyD1 were remarkably increased at 72 h after reperfusion. (Mean ± SD; #p<0.05; *p<0.01; NS- nonsignificant.)

Mentions: The promoter of CyD1 gene contains an AP-1 site. Therefore EMSA assay was used to assess the formation of AP-1/DNA complexes. As is shown, although the AP-1/DNA complexes in both groups were nearly similar at 1 and 6 h after reperfusion, it showed a significant increase after 24 h in the MSCs therapy group (Fig 7A). On the other hand, activation of NF-κB was identified at 1 h after reperfusion in both groups. However its activity in the 30PLT+MSCs group was 1.4-fold of that in the 30PLT+PBS group. Furthermore, it maintained high activity after 24 h, when activity of NF-κB in the 30PLT+MSCs group was 2.1-fold of that in the 30PLT+PBS group (Fig. 7A).


Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts: involvement of C-Jun N-terminal kinase, cyclin D1, and NF-κB.

Wang W, Du Z, Yan J, Ma D, Shi M, Zhang M, Peng C, Li H - PLoS ONE (2014)

MSCs therapy kept the activity of transcription factors AP-1 and NF-κB, and increased the expressions of phosphorylated c-Jun (p-c-Jun) and Cyclin D1 (CyD1).A. Activation of NF-κB and AP-1 was evaluated by electrophoretic mobility shift assay at 1, 6 and 24 h after reperfusion. Compared with the 30PLT+PBS group, the activity of NF-κB and AP-1 was considerably enhanced at 24 h after reperfusion in the 30PLT+MSCs group; B. Expressions of p-c-Jun and CyD1 were detected by western blotting at 6 h, 24 h, 72 h, and 7 days after reperfusion. Expressions of p-c-Jun and CyD1 were remarkably increased at 72 h after reperfusion. (Mean ± SD; #p<0.05; *p<0.01; NS- nonsignificant.)
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4257551&req=5

pone-0112532-g007: MSCs therapy kept the activity of transcription factors AP-1 and NF-κB, and increased the expressions of phosphorylated c-Jun (p-c-Jun) and Cyclin D1 (CyD1).A. Activation of NF-κB and AP-1 was evaluated by electrophoretic mobility shift assay at 1, 6 and 24 h after reperfusion. Compared with the 30PLT+PBS group, the activity of NF-κB and AP-1 was considerably enhanced at 24 h after reperfusion in the 30PLT+MSCs group; B. Expressions of p-c-Jun and CyD1 were detected by western blotting at 6 h, 24 h, 72 h, and 7 days after reperfusion. Expressions of p-c-Jun and CyD1 were remarkably increased at 72 h after reperfusion. (Mean ± SD; #p<0.05; *p<0.01; NS- nonsignificant.)
Mentions: The promoter of CyD1 gene contains an AP-1 site. Therefore EMSA assay was used to assess the formation of AP-1/DNA complexes. As is shown, although the AP-1/DNA complexes in both groups were nearly similar at 1 and 6 h after reperfusion, it showed a significant increase after 24 h in the MSCs therapy group (Fig 7A). On the other hand, activation of NF-κB was identified at 1 h after reperfusion in both groups. However its activity in the 30PLT+MSCs group was 1.4-fold of that in the 30PLT+PBS group. Furthermore, it maintained high activity after 24 h, when activity of NF-κB in the 30PLT+MSCs group was 2.1-fold of that in the 30PLT+PBS group (Fig. 7A).

Bottom Line: MSCs were successfully induced into hepatocyte-like cells, osteoblasts and adipocytes in vitro.Increased activities of AP-1 and NF-κB, as well as elevated expressions of p-c-Jun, cyclin D1, and proliferating cell nuclear antigen (PCNA), were also found in MSCs therapy group.These data suggest that MSCs therapy promotes hepatocyte proliferation and prolongs survival in SFSLT by reducing ischemia reperfusion injury and acute inflammation, and sustaining early increased expressions of c-Jun N-terminal Kinase, Cyclin D1, and NF-κB.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China; Department of Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou University School of Medicine, Zhengzhou, Henan Province, China.

ABSTRACT

Background: The therapeutic potential of mesenchymal stem cells (MSCs) has been highlighted recently for treatment of acute or chronic liver injury, by possibly differentiating into hepatocyte-like cells, reducing inflammation, and enhancing tissue repair. Despite recent progress, exact mechanisms of action are not clearly elucidated. In this study, we attempted to explore whether and how MSCs protected hepatocytes and stimulated allograft regeneration in small-for-size liver transplantation (SFSLT).

Methods: SFSLT model was established with a 30% partial liver transplantation (30PLT) in rats. The differentiation potential and characteristics of bone marrow derived MSCs were explored in vitro. MSCs were infused transvenously immediately after graft implantation in therapy group. Expressions of apoptosis-, inflammatory-, anti-inflammatory-, and growth factor-related genes were measured by RT-PCR, activities of transcription factors AP-1 and NF-κB were analyzed by EMSA, and proliferative responses of the hepatic graft were evaluated by immunohistochemistry and western blot.

Results: MSCs were successfully induced into hepatocyte-like cells, osteoblasts and adipocytes in vitro. MSCs therapy could not only alleviate ischemia reperfusion injury and acute inflammation to promote liver regeneration, but also profoundly improve one week survival rate. It markedly up-regulated the mRNA expressions of HGF, Bcl-2, Bcl-XL, IL-6, IL-10, IP-10, and CXCR2, however, down-regulated TNF-α. Increased activities of AP-1 and NF-κB, as well as elevated expressions of p-c-Jun, cyclin D1, and proliferating cell nuclear antigen (PCNA), were also found in MSCs therapy group.

Conclusion: These data suggest that MSCs therapy promotes hepatocyte proliferation and prolongs survival in SFSLT by reducing ischemia reperfusion injury and acute inflammation, and sustaining early increased expressions of c-Jun N-terminal Kinase, Cyclin D1, and NF-κB.

Show MeSH
Related in: MedlinePlus