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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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Assessment of liver fibrosis in MSC/HGF-transplanted rats and controls. (a) Diagram of the treatment protocol. (b) Extracellular deposition of collagen fibers stained with Sirius Red. Scale bar=0.5 mm. (c) Quantification of collagen by image analysis. (d, e) Collagen content and body weight quantified before (white bar) and 12 days after transplantation (black bar). (f) Hydroxyproline content. Normal rats (N), control fibrotic animals (S), fibrotic animals transplanted with MSCs (M) or MSCs/HGF (H). Data represent mean±s.d. for each group (*P<0.05, **P<0.01, ***P<0.005).
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fig2: Assessment of liver fibrosis in MSC/HGF-transplanted rats and controls. (a) Diagram of the treatment protocol. (b) Extracellular deposition of collagen fibers stained with Sirius Red. Scale bar=0.5 mm. (c) Quantification of collagen by image analysis. (d, e) Collagen content and body weight quantified before (white bar) and 12 days after transplantation (black bar). (f) Hydroxyproline content. Normal rats (N), control fibrotic animals (S), fibrotic animals transplanted with MSCs (M) or MSCs/HGF (H). Data represent mean±s.d. for each group (*P<0.05, **P<0.01, ***P<0.005).

Mentions: We adopted the DMN-induced rat liver fibrosis model in this study rather than the CCl4-induced model because CCl4-induced liver fibrosis resolves spontaneously.25 To address the therapeutic effect rather than the preventive effect, we injected saline, MSCs, or MSCs/HGF into the spleens of the rats once on the ninth day after the final DMN injection (4 days after biopsy; Figure 2a). The average collagen content (Figure 2c) and average body weight (Figure 2e) of all DMN-treated animals were similar at the time of cell transplantation, indicating that the extent of fibrosis was not different before transplantation. The rats were killed at 12 days after cell transplantation, and fibrosis was evaluated. Liver fibrosis was evident based on the formation of fibrotic septa joining the central area (Figure 2b, right upper panel). Quantitative measurement of collagen in liver tissue by Sirius red assay also revealed increased collagen content (Figure 2d, normal, 0.506±0.019 vs saline/DMN, 0.748±0.018, P<0.001), which was consistent with the histological changes. The increased collagen content of the rat livers was sustained during the entire experimental period after completion of DMN conditioning, indicating that the fibrosis in this experimental system was stable (Figure 2d, liver biopsy, 0.748±0.018 vs killing, 0.751±0.035 in the saline/DMN group).


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)

Assessment of liver fibrosis in MSC/HGF-transplanted rats and controls. (a) Diagram of the treatment protocol. (b) Extracellular deposition of collagen fibers stained with Sirius Red. Scale bar=0.5 mm. (c) Quantification of collagen by image analysis. (d, e) Collagen content and body weight quantified before (white bar) and 12 days after transplantation (black bar). (f) Hydroxyproline content. Normal rats (N), control fibrotic animals (S), fibrotic animals transplanted with MSCs (M) or MSCs/HGF (H). Data represent mean±s.d. for each group (*P<0.05, **P<0.01, ***P<0.005).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: Assessment of liver fibrosis in MSC/HGF-transplanted rats and controls. (a) Diagram of the treatment protocol. (b) Extracellular deposition of collagen fibers stained with Sirius Red. Scale bar=0.5 mm. (c) Quantification of collagen by image analysis. (d, e) Collagen content and body weight quantified before (white bar) and 12 days after transplantation (black bar). (f) Hydroxyproline content. Normal rats (N), control fibrotic animals (S), fibrotic animals transplanted with MSCs (M) or MSCs/HGF (H). Data represent mean±s.d. for each group (*P<0.05, **P<0.01, ***P<0.005).
Mentions: We adopted the DMN-induced rat liver fibrosis model in this study rather than the CCl4-induced model because CCl4-induced liver fibrosis resolves spontaneously.25 To address the therapeutic effect rather than the preventive effect, we injected saline, MSCs, or MSCs/HGF into the spleens of the rats once on the ninth day after the final DMN injection (4 days after biopsy; Figure 2a). The average collagen content (Figure 2c) and average body weight (Figure 2e) of all DMN-treated animals were similar at the time of cell transplantation, indicating that the extent of fibrosis was not different before transplantation. The rats were killed at 12 days after cell transplantation, and fibrosis was evaluated. Liver fibrosis was evident based on the formation of fibrotic septa joining the central area (Figure 2b, right upper panel). Quantitative measurement of collagen in liver tissue by Sirius red assay also revealed increased collagen content (Figure 2d, normal, 0.506±0.019 vs saline/DMN, 0.748±0.018, P<0.001), which was consistent with the histological changes. The increased collagen content of the rat livers was sustained during the entire experimental period after completion of DMN conditioning, indicating that the fibrosis in this experimental system was stable (Figure 2d, liver biopsy, 0.748±0.018 vs killing, 0.751±0.035 in the saline/DMN group).

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