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Therapeutic effect of hepatocyte growth factor-secreting mesenchymal stem cells in a rat model of liver fibrosis.

Kim MD, Kim SS, Cha HY, Jang SH, Chang DY, Kim W, Suh-Kim H, Lee JH - Exp. Mol. Med. (2014)

Bottom Line: Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy.Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced.In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry, Ajou University School of Medicine, Suwon, Republic of Korea [2] Chronic Inflammatory Disease Research Center, Ajou University School of Medicine, Suwon, Republic of Korea.

ABSTRACT
Bone marrow-derived mesenchymal stromal cells (MSCs) have been reported to be beneficial for the treatment of liver fibrosis. Here, we investigated the use of genetically engineered MSCs that overexpress hepatocyte growth factor (HGF) as a means to improve their therapeutic effect in liver fibrosis. Liver fibrosis was induced by intraperitoneal injection of dimethylnitrosamine. HGF-secreting MSCs (MSCs/HGF) were prepared by transducing MSCs with an adenovirus carrying HGF-encoding cDNA. MSCs or MSCs/HGF were injected directly into the spleen of fibrotic rats. Tissue fibrosis was assessed by histological analysis 12 days after stem cell injection. Although treatment with MSCs reduced fibrosis, treatment with MSCs/HGF produced a more significant reduction and was associated with elevated HGF levels in the portal vein. Collagen levels in the liver extract were decreased after MSC/HGF therapy, suggesting recovery from fibrosis. Furthermore, liver function was improved in animals receiving MSCs/HGF, indicating that MSC/HGF therapy resulted not only in reduction of liver fibrosis but also in improvement of hepatocyte function. Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy. Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced. In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF. Thus, MSC/HGF represents a promising approach toward a cell therapy for liver fibrosis.

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Analysis of human HGF expression in rats transplanted with MSCs/HGF. (a) Human HGF proteins. Values represent the mean±s.d. of triplicate assays of individual rats. (b) Human HGF mRNA. (c) Immunostaining for human mitochondria-specific antigen (hMT) and human HGF (hHGF). Blue: bis-benzamide, green: hMT, red: hHGF, Scale bar, 50 μm. Samples are fibrotic livers from control animals or from animals transplanted with MSCs or MSCs/HGF except in (a), where analysis was performed on serum collected from the portal vein 12 days after transplantation.
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fig4: Analysis of human HGF expression in rats transplanted with MSCs/HGF. (a) Human HGF proteins. Values represent the mean±s.d. of triplicate assays of individual rats. (b) Human HGF mRNA. (c) Immunostaining for human mitochondria-specific antigen (hMT) and human HGF (hHGF). Blue: bis-benzamide, green: hMT, red: hHGF, Scale bar, 50 μm. Samples are fibrotic livers from control animals or from animals transplanted with MSCs or MSCs/HGF except in (a), where analysis was performed on serum collected from the portal vein 12 days after transplantation.

Mentions: Our model system was based on the xenotransplantation of human MSCs into rats in the absence of immunosuppression. Thus, we verified the presence of transplanted human cells and the production of human HGF at the time of killing. Because we injected the cells into the spleen, we assumed that the transplanted cells would act directly in the liver after migration or would act indirectly by secreting HGF into the bloodstream, which would eventually reach the liver via the portal vein. The human HGF level in the portal vein was measured by human HGF-specific ELISA. As shown in Figure 4a, human HGF was detected in eight of nine rats of the MSC/HGF group, and in only one of seven rats in the MSC group. In addition, the average level of HGF in the portal vein was significantly higher in the MSC/HGF group than in the MSC group (MSC, 0.01±0.018 ng ml−1 vs MSC/HGF, 0.19±0.154 ng ml−1), indicating that a considerable amount of human HGF produced by cells transplanted into the spleen was delivered to the liver during the experimental period.


Therapeutic effect of hepatocyte growth factor-secreting mesenchymal stem cells in a rat model of liver fibrosis.

Kim MD, Kim SS, Cha HY, Jang SH, Chang DY, Kim W, Suh-Kim H, Lee JH - Exp. Mol. Med. (2014)

Analysis of human HGF expression in rats transplanted with MSCs/HGF. (a) Human HGF proteins. Values represent the mean±s.d. of triplicate assays of individual rats. (b) Human HGF mRNA. (c) Immunostaining for human mitochondria-specific antigen (hMT) and human HGF (hHGF). Blue: bis-benzamide, green: hMT, red: hHGF, Scale bar, 50 μm. Samples are fibrotic livers from control animals or from animals transplanted with MSCs or MSCs/HGF except in (a), where analysis was performed on serum collected from the portal vein 12 days after transplantation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150933&req=5

fig4: Analysis of human HGF expression in rats transplanted with MSCs/HGF. (a) Human HGF proteins. Values represent the mean±s.d. of triplicate assays of individual rats. (b) Human HGF mRNA. (c) Immunostaining for human mitochondria-specific antigen (hMT) and human HGF (hHGF). Blue: bis-benzamide, green: hMT, red: hHGF, Scale bar, 50 μm. Samples are fibrotic livers from control animals or from animals transplanted with MSCs or MSCs/HGF except in (a), where analysis was performed on serum collected from the portal vein 12 days after transplantation.
Mentions: Our model system was based on the xenotransplantation of human MSCs into rats in the absence of immunosuppression. Thus, we verified the presence of transplanted human cells and the production of human HGF at the time of killing. Because we injected the cells into the spleen, we assumed that the transplanted cells would act directly in the liver after migration or would act indirectly by secreting HGF into the bloodstream, which would eventually reach the liver via the portal vein. The human HGF level in the portal vein was measured by human HGF-specific ELISA. As shown in Figure 4a, human HGF was detected in eight of nine rats of the MSC/HGF group, and in only one of seven rats in the MSC group. In addition, the average level of HGF in the portal vein was significantly higher in the MSC/HGF group than in the MSC group (MSC, 0.01±0.018 ng ml−1 vs MSC/HGF, 0.19±0.154 ng ml−1), indicating that a considerable amount of human HGF produced by cells transplanted into the spleen was delivered to the liver during the experimental period.

Bottom Line: Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy.Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced.In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry, Ajou University School of Medicine, Suwon, Republic of Korea [2] Chronic Inflammatory Disease Research Center, Ajou University School of Medicine, Suwon, Republic of Korea.

ABSTRACT
Bone marrow-derived mesenchymal stromal cells (MSCs) have been reported to be beneficial for the treatment of liver fibrosis. Here, we investigated the use of genetically engineered MSCs that overexpress hepatocyte growth factor (HGF) as a means to improve their therapeutic effect in liver fibrosis. Liver fibrosis was induced by intraperitoneal injection of dimethylnitrosamine. HGF-secreting MSCs (MSCs/HGF) were prepared by transducing MSCs with an adenovirus carrying HGF-encoding cDNA. MSCs or MSCs/HGF were injected directly into the spleen of fibrotic rats. Tissue fibrosis was assessed by histological analysis 12 days after stem cell injection. Although treatment with MSCs reduced fibrosis, treatment with MSCs/HGF produced a more significant reduction and was associated with elevated HGF levels in the portal vein. Collagen levels in the liver extract were decreased after MSC/HGF therapy, suggesting recovery from fibrosis. Furthermore, liver function was improved in animals receiving MSCs/HGF, indicating that MSC/HGF therapy resulted not only in reduction of liver fibrosis but also in improvement of hepatocyte function. Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy. Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced. In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF. Thus, MSC/HGF represents a promising approach toward a cell therapy for liver fibrosis.

Show MeSH
Related in: MedlinePlus