Depressive Effect Of Shenfu Injection On Apoptosis Of Intestinal Mucosal Epithelial Cells And Its Potential Mechanism During Ischemia-Reperfusion In Rats

Objective: To investigate the effect of ShenFu injection on apoptosis of intestinal mucosal epithelial cells and its protective mechanism during ischemia-reperfusion in rats.Methods: Twenty-four SD rats weighing 250 25g were divided randomly into control group (group C), ischemia-reperfusion group (group I/R) and ShenFu injection group (group SF). The animals were fasted for 12 hours before commencing the experiments but had free access to water. Urethane (1mg/kg) was administered into the intraperitoneal cavity to induce anaesthesia. Through a midline abdominal incision, the superior mesenteric artery (SMA) was occluded for 60 minutes with an atraumatic vascular clamp and followed by reperfusion for 120 minutes in I/R group and SF group. The control animals underwent the same procedure except for the clamping of the SMA. ShenFu injection was injected 30 minetes before occlusion in SF group. 0.9% Natrii chloride was injected in I/R group and C group at the same time. Oxygen was inbreathed during the operation and Ischemia-reperfusion; At chosed time 3cm of jejunum of rate were harvested to sections for HE, Tunnel staining and caspase-3, Bcl-2 , Fas protein immunohistochemical taining. The sample was also take for the measurement of TNF-a with Enzyme-linked immunoadsordent assay (ELISA).Results: (1) The numble of apoptosis cells increased notably in group I/R, the numble of apoptosis cells was significantly lowed in group SF than in group I/R, while it was higher than that in group C (P<0.05). (2) The expression of Caspase-3 in group I/R increased notably and was higher than that in group C and group SF (P<0.01). There wasn't distinctly difference between group C and group SF in the expression of Caspase-3; There was positive correlation between the expression of Caspase-3 and apoptosis index in group I/R and in group SF (r=0.915, P<0.01; r=0.853, P<0.01). (3) The expression of Fas in group I/R increased notably and was higher than that in group C and group SF (P<0.01). There wasn't distinctly difference between group C and group SF in the expression of Caspase-3; The expression of Bcl-2 in group SF was markedly higher than that in group I/R (P<0.05), the expression of Bcl-2 in I/R group was less than that in group C (P<0.01). There was positive correlation between the expression of Fas and apoptosis index in group I/R and in group SF (r=0.669, P<0.05; r=0.724, P<0.05). There was negative correlation between the expression of Caspase-3 and Bcl-2 in group I/R and in group SF (r=-0.938, P<0.01; r=-0.916, P<0.01). (4)Thelever of TNF-ct in I/R group was markedly higher than that in group C and group SF (P<0.05), but there wasn't significantly difference between group C and group SF (P<0.01). There was positive correlation between the lever of TNF-a and apoptosis index in group I/R and in group SF (r=0.924, P<0.01; r=0.873, P<0.01). (5)The histopathology of intestinal mucosal was significantly better in group SF than in group I/R.Conclusions: Shenfu injection has depressive effect on the apoptosis of intestinal mucosal epithelial cells during ischemia-reperfusion by restraining the generation and releasing of TNF-a, down-regulating the expression of Caspase-3 , Fas and up-regulating the expression of Bcl-2, thus to alleviate the injury of intestinal mucosal epithelial cells during ischemia-reperfusion.