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Hepatocyte growth factor gene-modified adipose-derived mesenchymal stem cells ameliorate radiation induced liver damage in a rat model.

Zhang J, Zhou S, Zhou Y, Feng F, Wang Q, Zhu X, Ai H, Huang X, Zhang X - PLoS ONE (2014)

Bottom Line: Two days after irradiation, a significant reduction in apoptosis was observed in the HGF-overexpressing ADSC group compared with the RILD group, as assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining.The effect of mitigating RILD was compromised in the ADSC + shHGF group compared with the ADSC group.Altogether, these results suggest that HGF-overexpressing ADSCs can significantly improve RILD in a rat model, which may serve as a valuable therapeutic alternative.

View Article: PubMed Central - PubMed

Affiliation: Peking University People's Hospital, Peking University Institute of Hematology, Xicheng District, Beijing, China.

ABSTRACT
Liver damage caused by radiotherapy is associated with a high mortality rate, but no established treatment exists. Adipose-derived mesenchymal stem cells (ADSCs) are capable of migration to injured tissue sites, where they aid in the repair of the damage. Hepatocyte growth factor (HGF) is critical for damage repair due to its anti-apoptotic, anti-fibrotic and cell regeneration-promoting effects. This study was performed to investigate the therapeutic effects of HGF-overexpressing ADSCs on radiation-induced liver damage (RILD). ADSCs were infected with a lentivirus encoding HGF and HGF-shRNA. Sprague-Dawley (SD) rats received 60Gy of irradiation to induce liver injury and were immediately given either saline, ADSCs, ADSCs + HGF or ADSCs + shHGF. Two days after irradiation, a significant reduction in apoptosis was observed in the HGF-overexpressing ADSC group compared with the RILD group, as assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Scanning electron microscopy showed chromatin condensation after irradiation, which was ameliorated in the group that received ADSCs and was reversed in the group that received HGF-overexpressing ADSCs. HGF-overexpressing ADSCs ameliorated radiation- induced liver fibrosis through down regulation of α-SMA and fibronectin. Hepatocyte regeneration was significantly improved in rats treated with ADSCs compared with rats from the RILD group), as assessed by Ki-67 immunohistochemistry. Rats that received HGF-overexpressing ADSCs showed an even greater level of hepatocyte regeneration. HGF-overexpressing ADSCs completely blocked the radiation-induced increase in the enzymes ALT and AST. The effect of mitigating RILD was compromised in the ADSC + shHGF group compared with the ADSC group. Altogether, these results suggest that HGF-overexpressing ADSCs can significantly improve RILD in a rat model, which may serve as a valuable therapeutic alternative.

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Histological examination of the liver 60 days after irradiation.(A) Histopathology of liver sections stained with hematoxylin-eosin (HE) (400×). (B) Masson's trichome stain of liver sections. (C) Expression of collagen-I in rat livers detected by western blot. (D) Quantification of collagen-I protein expression detected by western blot, *p<0.05. (E) Histopathology of liver sections immunostained with antibodies to fibronectin (FN) and α-SMA (400×). Arrows indicated positive stain of fibronectin (in the images of FN) and α-SMA (in the images of α-SMA).
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pone-0114670-g004: Histological examination of the liver 60 days after irradiation.(A) Histopathology of liver sections stained with hematoxylin-eosin (HE) (400×). (B) Masson's trichome stain of liver sections. (C) Expression of collagen-I in rat livers detected by western blot. (D) Quantification of collagen-I protein expression detected by western blot, *p<0.05. (E) Histopathology of liver sections immunostained with antibodies to fibronectin (FN) and α-SMA (400×). Arrows indicated positive stain of fibronectin (in the images of FN) and α-SMA (in the images of α-SMA).

Mentions: Because radiation induces fibrosis in the liver, we performed fibrosis detection assays to determine whether ADSCs that overexpress HGF can mitigate fibrotic changes induced by the radiation. Histological examination of liver sections from RILD animals showed liver congestion and hepatocellular ballooning. All three groups that were treated with ADSCs showed amelioration in hepatocellular ballooning. HGF gene-modified ADSCs even abrogated the vacuolar degeneration caused by the irradiation (Fig. 4A). To demonstrate the extracellular collagen deposits in the liver tissue, we performed Masson's trichome staining. Our results demonstrate that radiation can induce collagen deposition in the portal and perisinusoidal regions. However, liver cirrhosis, which is characterized by the formation of a pseudolobule, was not observed 60 days after irradiation. Compared to that of the RILD group, the collagen deposit in the ADSC group was reduced. Almost no collagen accumulation was observed in the lenti-HGF + ADSCs groups (Fig. 4B). Western blot was used to quantify the protein expression (Fig. 4C). A significant up-regulation of collagen-I protein expression was observed in rats from the RILD group after irradiation (0.33±0.09 vs. 0.06±0.07, p<0.05), but this protein was significantly reduced in rats from the lenti-HGF + ADSC group (0.12±0.07 vs. 0.33±0.09, p<0.05) (Fig. 4D). The activation of hepatic stellate cells is a major event that is associated with the process of liver fibrosis, and α-SMA is up-regulated during the activation of these cells. As revealed in Fig. 4E, there were few a-SMA positive regions in the control group. In contrast, considerable a-SMA-positive regions can be seen around the perisinusoidal areas in the RILD group, whereas the treatment of HGF over-expressing ADSCs attenuated the expression of a-SMA and fibronectin compared to that of the RILD group. Immunohistochemical staining for fibronectin revealed that radiation-induced liver fibrosis was mitigated by the administration of ADSCs and lenti-HGF + ADSCs (Fig. 4E).


Hepatocyte growth factor gene-modified adipose-derived mesenchymal stem cells ameliorate radiation induced liver damage in a rat model.

Zhang J, Zhou S, Zhou Y, Feng F, Wang Q, Zhu X, Ai H, Huang X, Zhang X - PLoS ONE (2014)

Histological examination of the liver 60 days after irradiation.(A) Histopathology of liver sections stained with hematoxylin-eosin (HE) (400×). (B) Masson's trichome stain of liver sections. (C) Expression of collagen-I in rat livers detected by western blot. (D) Quantification of collagen-I protein expression detected by western blot, *p<0.05. (E) Histopathology of liver sections immunostained with antibodies to fibronectin (FN) and α-SMA (400×). Arrows indicated positive stain of fibronectin (in the images of FN) and α-SMA (in the images of α-SMA).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4264768&req=5

pone-0114670-g004: Histological examination of the liver 60 days after irradiation.(A) Histopathology of liver sections stained with hematoxylin-eosin (HE) (400×). (B) Masson's trichome stain of liver sections. (C) Expression of collagen-I in rat livers detected by western blot. (D) Quantification of collagen-I protein expression detected by western blot, *p<0.05. (E) Histopathology of liver sections immunostained with antibodies to fibronectin (FN) and α-SMA (400×). Arrows indicated positive stain of fibronectin (in the images of FN) and α-SMA (in the images of α-SMA).
Mentions: Because radiation induces fibrosis in the liver, we performed fibrosis detection assays to determine whether ADSCs that overexpress HGF can mitigate fibrotic changes induced by the radiation. Histological examination of liver sections from RILD animals showed liver congestion and hepatocellular ballooning. All three groups that were treated with ADSCs showed amelioration in hepatocellular ballooning. HGF gene-modified ADSCs even abrogated the vacuolar degeneration caused by the irradiation (Fig. 4A). To demonstrate the extracellular collagen deposits in the liver tissue, we performed Masson's trichome staining. Our results demonstrate that radiation can induce collagen deposition in the portal and perisinusoidal regions. However, liver cirrhosis, which is characterized by the formation of a pseudolobule, was not observed 60 days after irradiation. Compared to that of the RILD group, the collagen deposit in the ADSC group was reduced. Almost no collagen accumulation was observed in the lenti-HGF + ADSCs groups (Fig. 4B). Western blot was used to quantify the protein expression (Fig. 4C). A significant up-regulation of collagen-I protein expression was observed in rats from the RILD group after irradiation (0.33±0.09 vs. 0.06±0.07, p<0.05), but this protein was significantly reduced in rats from the lenti-HGF + ADSC group (0.12±0.07 vs. 0.33±0.09, p<0.05) (Fig. 4D). The activation of hepatic stellate cells is a major event that is associated with the process of liver fibrosis, and α-SMA is up-regulated during the activation of these cells. As revealed in Fig. 4E, there were few a-SMA positive regions in the control group. In contrast, considerable a-SMA-positive regions can be seen around the perisinusoidal areas in the RILD group, whereas the treatment of HGF over-expressing ADSCs attenuated the expression of a-SMA and fibronectin compared to that of the RILD group. Immunohistochemical staining for fibronectin revealed that radiation-induced liver fibrosis was mitigated by the administration of ADSCs and lenti-HGF + ADSCs (Fig. 4E).

Bottom Line: Two days after irradiation, a significant reduction in apoptosis was observed in the HGF-overexpressing ADSC group compared with the RILD group, as assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining.The effect of mitigating RILD was compromised in the ADSC + shHGF group compared with the ADSC group.Altogether, these results suggest that HGF-overexpressing ADSCs can significantly improve RILD in a rat model, which may serve as a valuable therapeutic alternative.

View Article: PubMed Central - PubMed

Affiliation: Peking University People's Hospital, Peking University Institute of Hematology, Xicheng District, Beijing, China.

ABSTRACT
Liver damage caused by radiotherapy is associated with a high mortality rate, but no established treatment exists. Adipose-derived mesenchymal stem cells (ADSCs) are capable of migration to injured tissue sites, where they aid in the repair of the damage. Hepatocyte growth factor (HGF) is critical for damage repair due to its anti-apoptotic, anti-fibrotic and cell regeneration-promoting effects. This study was performed to investigate the therapeutic effects of HGF-overexpressing ADSCs on radiation-induced liver damage (RILD). ADSCs were infected with a lentivirus encoding HGF and HGF-shRNA. Sprague-Dawley (SD) rats received 60Gy of irradiation to induce liver injury and were immediately given either saline, ADSCs, ADSCs + HGF or ADSCs + shHGF. Two days after irradiation, a significant reduction in apoptosis was observed in the HGF-overexpressing ADSC group compared with the RILD group, as assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Scanning electron microscopy showed chromatin condensation after irradiation, which was ameliorated in the group that received ADSCs and was reversed in the group that received HGF-overexpressing ADSCs. HGF-overexpressing ADSCs ameliorated radiation- induced liver fibrosis through down regulation of α-SMA and fibronectin. Hepatocyte regeneration was significantly improved in rats treated with ADSCs compared with rats from the RILD group), as assessed by Ki-67 immunohistochemistry. Rats that received HGF-overexpressing ADSCs showed an even greater level of hepatocyte regeneration. HGF-overexpressing ADSCs completely blocked the radiation-induced increase in the enzymes ALT and AST. The effect of mitigating RILD was compromised in the ADSC + shHGF group compared with the ADSC group. Altogether, these results suggest that HGF-overexpressing ADSCs can significantly improve RILD in a rat model, which may serve as a valuable therapeutic alternative.

Show MeSH
Related in: MedlinePlus