Figure 1.
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.
Figure 2.
HGF gene-modified ADSCs migrate to damaged tissue sites.
(A) TUNEL staining of liver sections from the control group and liver sections from the RILD group 2 days post-irradiation. Arrows indicate hepatocytes that are positive for TUNEL staining. (B) Quantification of TUNEL-positive cells. Five fields were randomly selected from each specimen, and the number of TUNEL-positive cells was determined (×400). (C) HE stain of liver specimens obtained from the control group and from the RILD group 60 days after irradiation. Arrow indicates vacuolar degeneration in liver. (D) ADSCs expressing GFP were detected in rat liver 7, 14 and 30 days after transplantation (100×). Arrows indicate cells positive for GFP staining. (E) HGF was detected by ELISA in rat liver 2 weeks after transplantation. *p<0.05.
Figure 3.
Evaluation of hepatocyte apoptosis 2 days after irradiation.
(A) Images of hepatocytes from an electron microscope (×8200). (B) TUNEL staining of liver specimens from all five groups (×400). (C) Quantification of cells that were positive for TUNEL staining: five random fields (200×) were selected from within each liver section, and TUNEL-positive cells were counted. Data are expressed as the mean ± SD. *p<0.05.
Figure 4.
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).
Figure 5.
Assessment of hepatocyte proliferation.
(A) Histological analysis of liver sections stained with a Ki-67 antibody (400×). Arrow indicates hepatocytes that are positive for Ki-67 staining. (B) Quantification of Ki-67-positive cells. Five random fields (400×) were selected from within each liver section, and the number of Ki-67-positive cells was counted. Data are expressed as the mean ± SD. *p<0.05.
Figure 6.
Analysis of liver function index.
Serum levels of ALT (A) and AST (B) 2 days after irradiation are shown. *p<0.05.