The Optimal Dosage of Bone Marrow Mesenchymal Stem Cells Transplantation for Treatment of Hepatic Ischemia-Reperfusion Injury in Rats_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

QIAO Pengfei , JIN Guangxin ,  WU Dequan

Department of General Surgery， The Second Affiliated Hospital of Harbin Medical University， Harbin 150086， Heilongjiang Province， China;

Corresponding author：

WU Dequan, Email: dqwu56@163.com

Keywords：

Bone marrow mesenchymal stem cells; Hepatic ischemia-reperfusion injury; Cell transplantation; Rat

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Abstract Full text Figures/Tables Video References Cited by

Objective 　To investigate the optimal dosage of bone marrow mesenchymal stem cells （BMSCs） transplantations for treatment of hepatic ischemia-reperfusion injury in rats， and to provide prophase experimental basis for it.
Methods 　BMSCs of Wistar rats were isolated and cultivated by bone marrow adherent culture method. BMSCs of the fourth generation were prepared for cell transplantation. Thrity hepatic ischemia-reperfusion injury models of male
Wistar rats were successfully established， and then were randomly divided into blank control group， 5×105 group， 1×106
group， 2×106 group， and 3×106 group， each group enrolled 6 rats. The 200 μL cell suspension of BMSCs were transfused
into the portal vein with number of 5×105， 1×106， 2×106， and 3×106 separately in rats of later 4 groups， and rats of blank control group were injected with phosphate buffered saline of equal volume. At 24 hours after cell transplantation， blood samples were collected to test aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， liver tissues
were obtained to test malonaldehyde （MDA）， superoxide dismutase （SOD）， and nuclear factor-κB （NF-κB） p65 protein.Liver tissues were also used to perform HE staining to observe the pathological changes.
Results 　Compared with blank control group， 5×105 group， and 3×106 group， the levels of AST， ALT， and MDA were lower （P＜0.05） while activity levels of SOD were higher （P＜0.05） in 1×106 group and 2×106 group， and expression levels of NF-κB p65 protein were lower with the pathological injury of liver tissue improved， but there were no significant differences on levels of AST， ALT， MDA， and SOD （P＞0.05）， and both of the 2 groups had the similar pathological change.
Conclusion　 The optimal dosage of the BMSCs transplantations after hepatic ischemia-reperfusion injury is 1×106.

Citation： QIAO Pengfei,JIN Guangxin,WU Dequan. The Optimal Dosage of Bone Marrow Mesenchymal Stem Cells Transplantation for Treatment of Hepatic Ischemia-Reperfusion Injury in Rats. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2013, 20(9): 1018-1022. doi: Copy

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11.	Zwacka RM， Zhang Y， Halldorson J， et al. CD4＋ T-lympho-cytes mediate ischemia/reperfusion-induced inflammatory responsesin mouse liver[J]. J Clin Invest， 1997， 100(2)：279-289.
12.	Jaeschke H， Bautista AP， Spolarics Z， et al. Superoxide generation by Kupffer cells and priming of neutrophils during reperfusion after hepatic ischemia[J]. Free Radic Res Commun， 1991， 15(5)：.
13.	Hernandez LA， Grisham MB， Twohig B， et al. Role of neutrophils in ischemia-reperfusion-induced microvascular injury[J]. Am J Physiol， 1987， 253 (3 Pt 2)：H699-H703.
14.	Orlic D， Kajstura J， Chimenti S， et al. Bone marrow cells regene-rate infarcted myocardium[J]. Nature， 2001， 410(6829)：701-705.
15.	Georgiev P， Dahm F， Graf R， et al. Blocking the path to death：anti-apoptotic molecules in ischemia/reperfusion injury of the liver[J]. Curr Pharm Des， 2006， 12(23)：2911-2921.
16.	Sen R， Baltimore D. Multiple nuclear factors interact with theimmunoglobulin enhancer sequences[J]. Cell， 1986， 46(5)：705-716.
17.	León Fernández OS， Ajamieh HH， Berlanga J， et al. Ozone oxid-ative preconditioning is mediated by A1 adenosine receptors in a rat model of liver ischemia/reperfusion[J]. Transpl Int， 2008， 21(1)：39-48.
18.	-284.
19.	， 8(4)：315-317.

1. Fan C， Zwacka RM， Engelhardt JF. Therapeutic approaches for ischemia/reperfusion injury in the liver[J]. J Mol Med (Berl)， 1999， 77(8)：577-592.
2. Kanazawa H， Fujimoto Y， Teratani T， et al. Bone marrow-derivedmesenchymal stem cells ameliorate hepatic ischemia reperfusion injury in a rat model[J]. PLoS One， 2011， 6(4)：e19195.
3. 金光鑫，乔鹏飞，胡彦华，等. 同种异体骨髓间充质干细胞保护大鼠肝脏热缺血再灌注损伤的研究[J]. 中国普外基础与临床杂志， 2013， 20(1)：60-65.
4. 石臣磊，秦华东，丁超. 依达拉奉对肝缺血再灌注损伤逆转作用的研究[J]. 中国普外基础与临床杂志， 2011， 18(8)：887-889.
5. Pittenger MF， Mackay AM， Beck SC， et al. Multilineage potential of adult human mesenchymal stem cells[J]. Science， 1999， 284(5411)：143-147.
6. Dominici M， Le Blanc K， Mueller I， et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement[J]. Cytotherapy，.
7. Casiraghi F， Azzollini N， Cassis P， et al. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells[J]. J Immunol， 2008， 181(6)：3933-3946.
8. Chabannes D， Hill M， Merieau E， et al. A role for heme oxyge-nase-1 in the immunosuppressive effect of adult rat and human mesenchymal stem cells[J]. Blood， 2007， 110(10)：3691-3694.
9. Popp FC， Eggenhofer E， Renner P， et al. Mesenchymal stem cells can induce long-term acceptance of solid organ allografts in synergy with low-dose mycophenolate[J]. Transpl Immunol， 2008， 20(1-2)：55-60.
10. Sbano P， Cuccia A， Mazzanti B， et al. Use of donor bone marrowmesenchymal stem cells for treatment of skin allograft rejection in a preclinical rat model[J]. Arch Dermatol Res， 2008， 300(3)：115-124.
11. Zwacka RM， Zhang Y， Halldorson J， et al. CD4＋ T-lympho-cytes mediate ischemia/reperfusion-induced inflammatory responsesin mouse liver[J]. J Clin Invest， 1997， 100(2)：279-289.
12. Jaeschke H， Bautista AP， Spolarics Z， et al. Superoxide generation by Kupffer cells and priming of neutrophils during reperfusion after hepatic ischemia[J]. Free Radic Res Commun， 1991， 15(5)：.
13. Hernandez LA， Grisham MB， Twohig B， et al. Role of neutrophils in ischemia-reperfusion-induced microvascular injury[J]. Am J Physiol， 1987， 253 (3 Pt 2)：H699-H703.
14. Orlic D， Kajstura J， Chimenti S， et al. Bone marrow cells regene-rate infarcted myocardium[J]. Nature， 2001， 410(6829)：701-705.
15. Georgiev P， Dahm F， Graf R， et al. Blocking the path to death：anti-apoptotic molecules in ischemia/reperfusion injury of the liver[J]. Curr Pharm Des， 2006， 12(23)：2911-2921.
16. Sen R， Baltimore D. Multiple nuclear factors interact with theimmunoglobulin enhancer sequences[J]. Cell， 1986， 46(5)：705-716.
17. León Fernández OS， Ajamieh HH， Berlanga J， et al. Ozone oxid-ative preconditioning is mediated by A1 adenosine receptors in a rat model of liver ischemia/reperfusion[J]. Transpl Int， 2008， 21(1)：39-48.
18. -284.
19. ， 8(4)：315-317.

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The Optimal Dosage of Bone Marrow Mesenchymal Stem Cells Transplantation for Treatment of Hepatic Ischemia-Reperfusion Injury in Rats

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