Combined mesenchymal stem cell transplantation and interleukin-1 receptor antagonism after partial hepatectomy.
OBJECTIVE
To study the therapeutic effects of mesenchymal stem cells (MSCs) and an interleukin-1 receptor antagonist (IL-1Ra) in acute liver failure.


METHODS
Chinese experimental miniature swine (15 ± 3 kg, 5-8 mo) were obtained from the Laboratory Animal Centre of the Affiliated Drum Tower Hospital of Nanjing University Medical School. Acute liver failure was induced via 85% hepatectomy, and animals were treated by MSC transplantation combined with IL-1Ra injection. Blood samples were collected for hepatic function analysis, and the living conditions and survival time were recorded. Liver injury was histologically analyzed. Hepatic cell regeneration and apoptosis were studied by Ki67 immunohistochemistry and terminal deoxynucleotidyl transferase dUTP nick end labeling, respectively. The levels of protein kinase B and nuclear factor-κB expression were analyzed by Western blotting.


RESULTS
MSCs were infected with a lentivirus for expression of green fluorescent protein (GFP) for subsequent identification; 97.3% of the MSCs were positive for GFP as assessed by flow cytometry. Additional flow cytometric analysis of cell surface marker expression demonstrated that > 90% of GFP-expressing MSCs were also positive for CD29, CD44, and CD90, indicating that most of these cells expressed typical markers of MSCs, and the population of MSCs was almost pure. Transplantation of MSCs in combination with 2 mg/kg IL-1Ra therapy significantly improved survival time compared to the acute liver failure model group (35.3 ± 6.7 d vs 17.3 ± 5.5 d, P < 0.05). Combined therapy also promoted improvement in serum inflammatory cytokines and biochemical conditions. The observed hepatic histopathologic score was significantly lower in the group with combined therapy than in the model group (3.50 ± 0.87 vs 8.17 ± 1.26, P < 0.01). In addition, liver cell apoptosis in the combined therapy group was significantly inhibited (18.1 ± 2.1% vs 70.8 ± 3.7%, P < 0.01), and hepatic cell regeneration increased. A significant increase in protein kinase B expression and decrease in nuclear factor-κB expression were observed (P < 0.01), which supports their important roles in liver regeneration.


CONCLUSIONS
MSCs and IL-1Ra had a synergistic effect in liver regeneration via regulation of inflammation and apoptotic signaling.
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Jian-Feng
JF
Sang
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Xiao-Lei
XL
Shi
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Bing
B
Han
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Xu
X
Huang
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Tao
T
Huang
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Hao-Zhen
HZ
Ren
Jian-Feng Sang, Xiao-Lei Shi, Bing Han, Xu Huang, Tao Huang, Hao-Zhen Ren, Yi-Tao Ding, Department of General Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China.
Yi-Tao
YT
Ding
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000097:blood; D004195:Disease Models, Animal; D006498:Hepatectomy; D018836:Inflammation Mediators / Q000097:blood; D053590:Interleukin 1 Receptor Antagonist Protein / Q000494:pharmacology*; D019394:Ki-67 Antigen / Q000378:metabolism; D008099:Liver / Q000000981:diagnostic imaging / Q000187:drug effects* / Q000378:metabolism / Q000601:surgery; D017114:Liver Failure, Acute / Q000097:blood / Q000175:diagnosis / Q000473:pathology / Q000628:therapy*; D008111:Liver Function Tests; D008115:Liver Regeneration / Q000187:drug effects*; D045164:Mesenchymal Stem Cell Transplantation; D016328:NF-kappa B / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D013556:Swine, Miniature; D013997:Time Factors; D014057:Tomography, X-Ray Computed; D014463:Ultrasonography
D016207:Cytokines; D018836:Inflammation Mediators; D053590:Interleukin 1 Receptor Antagonist Protein; D019394:Ki-67 Antigen; D016328:NF-kappa B; D051057:Proto-Oncogene Proteins c-akt
Acute liver failure; Hepatoectomy; Interleukin-1 receptor antagonist; Mesenchymal stem cells; Stem cell transplantation