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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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Effect of MSC/HGF transplantation on Ito cell activation. (a) α-SMA and desmin were detected by immunohistochemistry in liver sections. Scale bar, 500 μm. (b) Liver extracts from each group were subjected to western blot analysis using antibodies against α-SMA and desmin. (c) The relative amount of α-SMA and desmin proteins was calculated by densitometry. Normal rats, N; control fibrotic animals, S; and fibrotic animals transplanted with MSCs, M, or MSCs/HGF, H. Data represent the mean±s.d. for each group. α-SMA; α-smooth muscle actin (*P<0.05, **P<0.01, ***P<0.005).
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fig6: Effect of MSC/HGF transplantation on Ito cell activation. (a) α-SMA and desmin were detected by immunohistochemistry in liver sections. Scale bar, 500 μm. (b) Liver extracts from each group were subjected to western blot analysis using antibodies against α-SMA and desmin. (c) The relative amount of α-SMA and desmin proteins was calculated by densitometry. Normal rats, N; control fibrotic animals, S; and fibrotic animals transplanted with MSCs, M, or MSCs/HGF, H. Data represent the mean±s.d. for each group. α-SMA; α-smooth muscle actin (*P<0.05, **P<0.01, ***P<0.005).

Mentions: HGF has been reported to directly affect the survival and activation of rat Ito cells in vivo.20 Therefore, we assessed the effect of MSCs/HGF and MSCs on Ito cells by measuring α-SMA, which is a marker of activated Ito cells, and desmin, which is a marker of Ito cells irrespective of their activation state.27 Immunohistochemistry revealed that α-SMA and desmin expression decreased more in the MSC/HGF group than in the MSC group (Figure 6a). Western blotting of pooled liver lysates from each group further confirmed this observation (Figure 6b). In addition, the calculated expression of α-SMA and desmin protein for each experimental animal was similar to that in pooled samples (Figure 6c). Based on these results, we conclude that MSCs/HGF decreased both the number and the activation of Ito cells to a greater extent than MSCs, most probably as a result of the known inhibitory activity of HGF on the survival and activation of Ito cells.


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)

Effect of MSC/HGF transplantation on Ito cell activation. (a) α-SMA and desmin were detected by immunohistochemistry in liver sections. Scale bar, 500 μm. (b) Liver extracts from each group were subjected to western blot analysis using antibodies against α-SMA and desmin. (c) The relative amount of α-SMA and desmin proteins was calculated by densitometry. Normal rats, N; control fibrotic animals, S; and fibrotic animals transplanted with MSCs, M, or MSCs/HGF, H. Data represent the mean±s.d. for each group. α-SMA; α-smooth muscle actin (*P<0.05, **P<0.01, ***P<0.005).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig6: Effect of MSC/HGF transplantation on Ito cell activation. (a) α-SMA and desmin were detected by immunohistochemistry in liver sections. Scale bar, 500 μm. (b) Liver extracts from each group were subjected to western blot analysis using antibodies against α-SMA and desmin. (c) The relative amount of α-SMA and desmin proteins was calculated by densitometry. Normal rats, N; control fibrotic animals, S; and fibrotic animals transplanted with MSCs, M, or MSCs/HGF, H. Data represent the mean±s.d. for each group. α-SMA; α-smooth muscle actin (*P<0.05, **P<0.01, ***P<0.005).
Mentions: HGF has been reported to directly affect the survival and activation of rat Ito cells in vivo.20 Therefore, we assessed the effect of MSCs/HGF and MSCs on Ito cells by measuring α-SMA, which is a marker of activated Ito cells, and desmin, which is a marker of Ito cells irrespective of their activation state.27 Immunohistochemistry revealed that α-SMA and desmin expression decreased more in the MSC/HGF group than in the MSC group (Figure 6a). Western blotting of pooled liver lysates from each group further confirmed this observation (Figure 6b). In addition, the calculated expression of α-SMA and desmin protein for each experimental animal was similar to that in pooled samples (Figure 6c). Based on these results, we conclude that MSCs/HGF decreased both the number and the activation of Ito cells to a greater extent than MSCs, most probably as a result of the known inhibitory activity of HGF on the survival and activation of Ito cells.

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