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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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Construction of HGF gene-modified ADSCs.(A) Micrographs of cultured ADSCs at passage three. (B) Surface antigens of ADSCs were analyzed by flow cytometry. Freshly cultured ADSCs were positive for CD90 (99%), but not for CD31 (0.02%), CD34 (0.02%), or CD45 (0.03%). (C) Infection efficiency 4 days post-infection. GFP-expressing ADSCs were detected and counted using fluorescence microscopy. The infection efficiency was 100%. (D) Expression of HGF mRNA as detected by RT-RCR 4days post-transfection. (E) Expression of HGF protein as detected by ELISA 4days post-transfection. *p<0.05.
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pone-0114670-g001: Construction of HGF gene-modified ADSCs.(A) Micrographs of cultured ADSCs at passage three. (B) Surface antigens of ADSCs were analyzed by flow cytometry. Freshly cultured ADSCs were positive for CD90 (99%), but not for CD31 (0.02%), CD34 (0.02%), or CD45 (0.03%). (C) Infection efficiency 4 days post-infection. GFP-expressing ADSCs were detected and counted using fluorescence microscopy. The infection efficiency was 100%. (D) Expression of HGF mRNA as detected by RT-RCR 4days post-transfection. (E) Expression of HGF protein as detected by ELISA 4days post-transfection. *p<0.05.

Mentions: After 4–5 hours in culture, a few dispersed cells were observed to be adherent on the bottom of the dish (Fig. 1A). Cells began to grow quickly after 3days and developed long spindly processes, occupied a small volume, and did not grow in any particular direction. The observations of cell morphology under a light microscope fit the characteristics of ADSCs. After the third passage, FACS analysis was used to verify the phenotype. The results showed that our cultured cells were positive for CD90 (99%) but were negative for expression of CD34 (0.02%), CD45 (0.03%) and CD31 (0.02%) (Fig. 1B).


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)

Construction of HGF gene-modified ADSCs.(A) Micrographs of cultured ADSCs at passage three. (B) Surface antigens of ADSCs were analyzed by flow cytometry. Freshly cultured ADSCs were positive for CD90 (99%), but not for CD31 (0.02%), CD34 (0.02%), or CD45 (0.03%). (C) Infection efficiency 4 days post-infection. GFP-expressing ADSCs were detected and counted using fluorescence microscopy. The infection efficiency was 100%. (D) Expression of HGF mRNA as detected by RT-RCR 4days post-transfection. (E) Expression of HGF protein as detected by ELISA 4days post-transfection. *p<0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0114670-g001: Construction of HGF gene-modified ADSCs.(A) Micrographs of cultured ADSCs at passage three. (B) Surface antigens of ADSCs were analyzed by flow cytometry. Freshly cultured ADSCs were positive for CD90 (99%), but not for CD31 (0.02%), CD34 (0.02%), or CD45 (0.03%). (C) Infection efficiency 4 days post-infection. GFP-expressing ADSCs were detected and counted using fluorescence microscopy. The infection efficiency was 100%. (D) Expression of HGF mRNA as detected by RT-RCR 4days post-transfection. (E) Expression of HGF protein as detected by ELISA 4days post-transfection. *p<0.05.
Mentions: After 4–5 hours in culture, a few dispersed cells were observed to be adherent on the bottom of the dish (Fig. 1A). Cells began to grow quickly after 3days and developed long spindly processes, occupied a small volume, and did not grow in any particular direction. The observations of cell morphology under a light microscope fit the characteristics of ADSCs. After the third passage, FACS analysis was used to verify the phenotype. The results showed that our cultured cells were positive for CD90 (99%) but were negative for expression of CD34 (0.02%), CD45 (0.03%) and CD31 (0.02%) (Fig. 1B).

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