L-Carnitine in Intestinal Ischemia/Reperfusion Injury J Clin Med Res • 2011;3(2):78-84 Table 2. The Levels of BT From MLN, Liver, Lung, Spleen and Portal Vein Blood (CFU/g) (Mean ± SD) Groups S IR IR+L n 10 10 10 MLN 1.2 ± 1.03Δ 25.1 ± 3.41* 11.3 ± 2.36 Δ P < 0.05 compared to IR and IR+L groups * P < 0.05 compared to IR+L group Liver 0.0 ± 0.00Δ 20.5 ± 3.03* 10.0 ± 2.58 croscope (Fig. 1). The bacteria were cultured for the study to the density of 7.5 × 106/ml. Intestinal IR injury model and operative procedures The thirty healthy SD rats were randomly divided into three groups (n = 10 for each group): sham operated group (S), IR group (IR) and IR+L-carnitine group (IR+L). All of the ani- mals were given the EmGFP labeled E. coli 2 ml by gavage two hours before surgical procedures. The rats were anes- thetized through injecting Urethane 200 mg/kg into the peri- toneal cavity. In the S group, the rats received a 3-centime- ter medium-length-wise laparotomy, and the small intestine was exposed. Subsequently, the superior mesenteric artery (SMA) was identiﬁed and dissected, and then the peritoneal cavity was closed. In the IR group, the rats underwent intes- tinal ischemia for 45 minutes through occlusion of the SMA with a micro vascular clamp. The occluding clamp was re- moved after ischemia for a reperfusion period for 2 hours. In the IR+L group, the rats were given intervention of intestinal ischemia, which was the same as that in the IR group, and they were injected L-caritine 80 mg/kg suspended in saline solution (total volume = 2 ml) through tail vein before re- perfusion injury. Meanwhile, the rats were given 2 ml saline solution instead of L-caritine in both the S and IR groups. Lung 0.0 ± 0.00Δ 25.5 ± 3.92* 10.8 ± 2.25 Spleen 0.0 ± 0.00Δ 15.2 ± 3.01* 10.3 ± 3.43 Portal vein blood 2.0 ± 2.58Δ 19.9 ± 3.70* 9.3 ± 1.64 At the end of the procedures, one milliliter venous sys- temic serum was collected in order to measure the levels of TNF-α, IL-1β, IL-6 and IL-10 in venous blood using ELISA. The rats were sacriﬁced and a one-centimeter segment of il- eum was dissected and processed using standard histologi- cal techniques including formalin ﬁxation, dehydration and parafﬁn embedding, and was subsequently cut into 4 μm sections and stained with Haematoxylin and Eosin. All of the sections were analyzed with an optic microscope by the pathologist who did not know which group each rat belonged to, and they were classiﬁed according to the degree of tissue injury in accordance to Chiu’s score classiﬁcation [14] (Ta- ble 1). The homogenate of the mesenteric lymph nodes, liv- er, spleen, lung and half milliliter blood of portal vein were detected for the BT. Among the homogenate, 0.1 gram was cultured in agar culture media with ampicillin (100 mg/L) at 37 °C for 24 hours in order to count the CFU/g of bacteria. Statistical analysis Data was presented as mean ± standard derivation (SD). One-way analysis of variance (ANOVA) and Tukey Post Hoc (parametric) test were used to compare serum cyto- kines and translocation of bacteria between the three groups. Histopathologic grades of each group were compared using Figure 2. Normal intestinal mucosa in the S group rats (Grade 0) (H&E, × 100). Figure 3. Intestinal villus after IR in group II rats, with lifted epithelium, white blood cells, and monocytes in the Lamina propria (Grade 4) (H&E, × 100). 80 Articles © The authors | Journal compilation © J Clin Med Res and Elmer Press™ | www.jocmr.org