Transcript L_ARG

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L-Arginine Modulate Intestinal Inflammation in Rats
Submitted to Mesenteric Ischemia-Reperfusion Injury
Ana Laura Aidar1, Leandro Ryuchi Iuamoto1, Eduardo Hiroshi Tikazawa1, Florense de Lucca Melo1, Karis de
Campos1, Thais de Melo Alexandre e Silva1, Guilherme dos Santos Perez1, Fernando Lopes Carrara1, Jordan
Bistafa Liu1, Jeferson Oliveira1, Hugo Pequeno Monteiro1, Djalma José Fagundes1,1, Murched Omar Taha1,1.
1Surgery Department, Sao Paulo Federal University, Sao Paulo, Brazil - email: [email protected]
INTRODUCTION
The ischemia (I) and reperfusion (R) promote intestinal lesions that alter the
enteric motor function and compromise the prognosis of intestinal
transplantation. Treatment with cytoprotective agents have been proposed to
increase the success of intestinal transplants. L-arginine is an amino acid
important precursor in the synthesis of nitric oxide and which is directly
involved in oxidative stress resulting from ischemia and reperfusion injury.
Thus interest is directed for the evaluation of the expression of genes that
encode proteins involved in the oxidative stress and antioxidant defense.
PURPOSE
To study whether treatment with the L-arginine attenuates intestinal
dysfunction caused by ischemia and reperfusion.
MATERIALS AND METHODS
Eighteen rats were used (n=6/group): Sham only submitted to surgery
proceeding; IR group: submitted to IR proceeding and treated with saline
solution (SS) and H+IR group: treated with Larg (100 mg/kg) injected into the
femoral vein 5 minute before I, 5 minutes before R, and 55 minutes after R. In
the end of experiment the animals were euthanized and the jejunal segments
(2 cm) were removed and washed, and the Polymerase Chain Reaction in
Real Time (real-time PCR) was performed. In summary the cDNA obtained in
the previous stage was used as template in the PCR reaction in real time using
the specific kits (Enteric Endothelial Cell Biology Biosciences - Qiagen, Co) for
each sample was analyzed by real time PCR to 84 genes.
RESULTS
Compared with SG group, pro-apoptotic gene Bax and anti-apoptotic gene
(Bcl2I2) were up-regulated.
AVG ΔCt
(Ct(GOI) - Ave Ct (HKG))
Fold Change
Test Sample
Control
Sample
Bax
-0.32
+1.94
Test Sample / p-value
Control
Sample
+4.78
0.000104
Blc2l2
-0.70
+2.81
+11.85
Symbol
T-TEST
0.000026
Table 1: Expression of two genes related to enteric endothelial call biology from rats of Sham ischemia and reperfusion (SS+IR) and
ischemic and reperfusion plus L-Arginina (Larg+IR) groups. Significant values of fold up (+) or down(-) regulation was marked in
bold [2^(-Delta Ct)]
CONCLUSIONS
The pretreatment with L-arginine in mesenteric ischemia-reperfusion injury was
demonstrated in rodents that could be related with ability of this drug to
increase the expression of pro-apoptotic gene Bax and anti-apoptotic gene
(Bcl2I2).