Studies On Prophylactic-therapeutic Effects Of Shen-Fu Injection On Intestinal Mucosa Injury After Ischemia/reperfusion In Rats

Objective: Shen-Fu injection has been effectively used in the treatment of different typies of ischemia-reperfusion injury.The purpose of this study was to explore the mechanism of its protective effect on intestinal ischemia reperfusion injury in rats.Methods: 36 Adult male Sprague-Dawley rats weighing 220-270g were anesthetized with ethyl urethane (1g/kg) and subjected to intestinal ischemia (1 hour of superior artery occlusion alone) followed by 2 hours of reperfusion. Animals were divided into three groups: (1)ischemia and reperfusion alone (n=12). (2)Shen-Fu injection pretreatment (10ml/kg 30min before ischemia, n=12).(3)sham operated0 controls (laparotomy, n=12). Pathologic methods were used to evaluate the degree of small intestine injury. Tumor necrosis factor-a (TNF-a) levels of plasma and small intestinal tissue in each group were measured by enzyme-linked immunoadsordent assay (ELISA), in addition, the expression levels and distribution of nuclear factor-KB (NF-KB), inducible nitric oxide synthase (iNOS) and intercellular adhesion molecule-1 (ICAM-1) in small intestinal tissues were determined by immunohistochemical technique respectively. The average optical density (OD) value of NF-KB, ICAM-1 and iNOS protein in each group were assessed through HIPAS-2000 image analysis system.Results: 1. The intestinal mucosa necrotic microstructure was very obvious in IR group. But in SF group, the intestinal mucosa microstructure was generally normal. The pathologic lesion of small intestine was significantly reduced in SF group compared with that in IR group (0.97 0.69 vs. 3.63 0.74; P<0.01). 2. The average OD value of NF-KB protein was significantly higher in IR group than that in SF group (0.1370 0.00823 vs. 0.1192 0.01541; P<0.01) and in C group (0.1370 0.00823 vs. 0.1010 0.02025; P<0.01). Compared with C group, IR increased the average OD value of ICAM-1 protein (0.2070 0.03945 vs. 0.0960 0.02716; P<0.01) and iNOS protein remarkably (0.1783 0.02563 vs. 0.1054 0.01014; P<0.01). In contrast to IR group, Shen-Fu injection decreased the average OD value of ICAM-1 protein (0.1150 0.02121 vs. 0.2070 0.03945; P<0.01) and iNOS protein significantly (0.1298 0.01865 vs. 0.1783 0.02563; P<0.01). The NF-KB protein was mostly located at cellular nuclei and sparsely at cellular plasm; the ICAM-1 and iNOS protein were located at cellular plasm. 3. The levels of TNF-a in the plasma and small intestinal tissue were obviously higher in IR group than those in SF group ((189.7 56.3)pg/ml and (1731 162)pg/g WT vs.(18.6 8.4)pg/ml and (38.6 10.4)pg/g WT; P<0.01).Conclusions: These result indicate that pretreatment with Shen-Fu injection can prevent gut ischemia reperfusion injury in rats, and such protective effect may be related to the inhibition of the activity of NF-KB and the expression of ICAM-1, iNOS and TNF-.