Floor plan - 2013 Annual Meeting - American Association for Hand ...

The Gene Expression Profiling of Ischemia-Reperfusion injury in Rat Kidney, Small Intestine,
and Cremasteric Muscle Model by DNA Microarray
Institution where the work was prepared: Chang Nai-Jen, Taoyuan, Taiwan
Nai-Jen Chang, yes; See-Tong Pang; Fu-Chan Wei; Chang-Gung Memorial Hospital
BACKGROUND:
Ischemia-reperfusion (I/R) injury is inevitable to cause tissue damage and leading to graft failure. Although susceptibility of various tissues
to I/R injury has been demonstrated, the detailed mechanism is not fully elucidated. DNA microarray is a powerful tool to detect
whole genome of the molecular changes during I/R injury. Here we compared the gene expression profile of I/R injury using kidney,
intestine, and cremasteric muscle model in rats.
METHODS:
45 male Lewis rats ranging from 270 to 330 gw were randomly assigned to one of 9 groups with an equal number in each group (n=5)
and prepared for the study. Left kidney, small intestine, and cremasteric muscle were prepared for I/R pedicle. In each model, the group
I is the time controlled group without ischemic insult. The group II had ischemic for 60 minutes. The group III had ischemic for 60 minutes
plus 60 minutes reperfusion. Extracted total RNA was used to probe the whole rat genome oligo array and the novel genes were
confirmed by real-time PCR.
RESULTS:
Gene expression profiling of the kidney, small intestine, and cremasteric muscle during I/R injury were performed. 23465 genes included
in our studies. After 60 minutes ischemia plus 60 minutes reperfusion, 654, 162, 776 genes were 2-folds up-regulated whereas 129,
518, 4837 genes 2-folds down-regulated in kidney, small intestine, and cremasteric muscle individually. There were no common gene
2-folds up-regulated in all three models during the reperfusion stage. Among the genes, we further identified activating protein-1(AP-
1), an important transcription factor. We found the expression of the AP-1 subunits were extremely high in renal and intestinal but downregulated
in muscle model, such as Fra-2 (18.75/17.33/0.24 folds), activating transcription factor 3 (ATF3) (13.26/5.77/0.26 folds).
CONCLUSION:
AP-1 expresses during cell stress and mediated in many biological pathways. The activation of AP-1 bases on the shifting conjugation
from [Jun]-[Jun] homodimer to [Jun]-[Fos] and [Jun]-[ATF3] heterodimers and activate the JNK and p38 ERK pathways (belong to MAPK
kinase pathway) individually. The dynamic balance of the two pathways may be important in determining whether a cell survives or
undergoes apoptosis. Ap-1 presented with up-regulation in kidney and intestine whereas down regulation after I/R insult may due to
the longer ischemic-tolerance of skeletal muscle. The specific role and the related pathway related to the I/R injury between these three
organs will be further analyzed.
Aged Donor Bone Marrow Influcences Mixed Chimerism and Donor-specific Tolerance to
Composite Tissue Allotransplantation with Nonmyeloablative Conditioning
Institution where the work was prepared: Chang Gung Memorial Hospital, Taipei, Taiwan
Jeng-Yee Lin, MD; Wei-Chao Huang; David C.C. Chuang; Fu-Chan Wei; Chang Gung Memorial Hospital
BACKGROUND:
Allogeneic transplantation with aged donor organ or tissue is associated with a higher acute rejection rate or delayed organ dysfunction.
The purpose of this study is to investigate if composite tissue allotransplantation (CTA) through mixed chimerism with aged donor
bone marrow transplantation faces a lower engraftment rate, lower mixed chimerism or higher CTA rejection rate.
MATERIAL/METHODS:
Twelve male (6-to-10-week old) and Lewis (LEW) rats (RTA1l) were equally categorized into two groups as recipients. Male Brown Norway
rats (RTA1c) were the donors. Group I recipients were transplanted with100 x 106 · ‚ -TCR+ and Á &delta-TCR+ T-cell depleted bone
marrow (BM) cells from 8-to-10-week-old Brown Norway (BN) rats. Group II recipients were transplanted with same number of T-cell
depleted BM cells from 14-to-16-month-old BN rats. Recipient rats were irradiated with 400cGy total body irradiation one day before
BMT and treated with one dose of 5 mg antilymphocyte serum (ALS) intraperitoneally (IP) one day before BMT, cyclosporine 16 mg/kg/d
from days 0 to 10 and one dose of 5 mg ALS, IP ten days after BMT. In both groups, the hindlimb osteomyocutaneous flap allotransplantation
(BN„? mixed chimera rat) was done 28 days after bone marrow transplantation (BN„?LEW). Chimerism level and multilineage
were assessed by flowcytometry 15, 30, 60, 120, and 150 days following BMT. The percentage of the facilitating cells (CD8+, · ‚-
TCR-, Á &delta -TCR-), and hemopoietic stem cells (SSClow, ALDFlourbright) in lymphoid gate were checked in donor BM cells.
RESULTS:
Recipients in both groups were 100 % engrafted with donor BM. The chimerism level in each group 28 days after BMT were as follows:
Group I: 50 %. Group II: 7%. The graft tolerance rate in Group I (80%) was significantly higher than in Group II (0%). The percentage of
facilitating cells in lymphoid gate were found to be higher in 6-to-10-week-old donor BM cells than in 12-to-16-month-old BM cells while
the percentage of hemopoietic stem cells were the same in two different age donor BM cells.
CONCLUSION:
Significantly lower mixed chimerism level and allograft tolerance rate were found in CTA through BMT with aged donor BM cells. Lower
mixed chiemrism level is probably associated with a lower percentage of facilitating cells rather than stem cells in the aged donor BM
cells.
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