| The First PartThe effects of hypertonic saline/hydroxyethyl starch solution combined with pentoxifylline on expresstion of inflammatory factor and function of intestinal mucosa in early resuscitation of severe hemorrhagic shock in ratsObjective To explore the influence of hypertonic saline/hydroxyethyl starch solution combined with pentoxifylline on expresstion of inflammatory factor and function of intestinal mucosa in the early resuscitation of severe hemorrhagic shock in rats.Methods 72 healthy male SD rats were randomly divided into six groups:control group (C), sham group (NR), Ringer's lactate group (RL), Ringer's lactate combined with pentoxifylline (RL/PTX), hypertonic saline/hydroxyethyl starch solution group (HSH) and hypertonic saline/hydroxyethyl starch solution combined with pentoxifylline group (HSH/PTX),12 rats in each group. The C group rats were only anesthetized and the right femoral artery and femoral vein catheter inserted, but no shock, no resuscitation. The other groups'rats were bled to remain a mean arterial pressure of (35±5) mmHg for 1h. According to Wigger's method, the rats'models of severe hemorrhagic shock were reproduced. The NR group rats were not resuscitated. The NR group rats were sacrificed at 1h after shock, and specimens were taken immediately after death to be tested. The RL group rats were given 3 times the amount of blood loss of RL which was constantly infused within 20 minutes. The RL/PTX group rats were given the same treatment as the RL group, but PTX (25mg/kg) was integrated into RL. The HSH group rats received HSH solution (6ml/kg) which was constantly infused within 5 minutes. The HSH/PTX group rats were given the same treatment as the HSH group, but PTX (25mg/kg) was integrated into HSH. Normal saline (6ml/kg·h) was constantly infused for 4 hours after fluid resuscitation in four fluid resuscitation groups. The mean arterial pressure (MAP) of rats was monitored before shock, before resuscitation and 0.5h, 1h,2h,3h,4h after resuscitation, respectivly. At the end of the experiment, the rats were sacrificed to measure the intestinal concentrations of IL-6, IL-10 and the plasma concentrations of TNF-a and i-FABP by ELISA. The expression of ICAM-1 in the intestinal tissue was detected by Immunohistochemical method. MPO activity in the intestinal tissue was detected by Spectrophotometric method. The damage degree of intestinal tissue was observed with Optical microscope. Epithelium, organelles connections, the structure and apoptosis in intestinal specimens was observed with transmission electronic microscope.Results 1.MAP changes after resuscitation:MAP of group RL, RL/PTX, HSH, HSH/PTX was increased obviously after resusucitation than that of before resuscitation (P<0.01). Compared with group RL and group RL/PTX, MAP was more stable in group HSH and group HSH/PTX (P<0.01).2.Inflammation factors change:Compared with group NR, group RL, group RL/PTX and group HSH, the pathological damage was lessened significantly, the levels of TNF-a, i-FABP in plasma and IL-6, ICAM-1, MPO in intestinal tissue were decreased, the concentration of IL-10 in intestinal tissue was increased significantly in group HSH/PTX (P<0.01).3. Optical microscope examination of intestinal tissue by hematoxylin-eosin (HE) staining, Chiu's damage score, electron ultrastructure pathology examination, i-FABP content in plasma:Compared with group NR, group RL, group RL/PTX and group HSH, Chiu's damage score and i-FABP content in plasma were reduced obviously in group HSH/PTX (P<0.01), the intestinal injury observed by optical microscope examination and electron ultrastructure pathology examination was milder in group HSH/PTX.Conclusion 1.HSH/PTX could effectively increase the MAP and keep hemodynamic stability in early fluid resuscitation of severe hemorrhagic shock in rats.2.HSH/PTX might relieve the injury of the intestinal mucous by decreasing the levels of TNF-a, IL-6, ICAM-1, i-FABP, increasing the concentration of IL-10 and reducing MPO activity and the expression of inflammatory regulatory factors ICAM-1. The second partThe effects of hypertonic saline/hydroxyethyl starch solution combined with pentoxifylline on oxidative stress damage in intestine tissue in early resuscitation of severe hemorrhagic shock in ratsObjective To explore the influence of hypertonic saline/hydroxyethyl starch solution combined with pentoxifylline on oxidatie stress damage in intestinal tissue in early resuscitation of severe hemorrhagic shock in rats.Methods 72 healthy male SD rats were randomly divided into six groups:control group (C), sham group (NR), Ringer's lactate group (RL), Ringer's lactate combined with pentoxifylline (RL/PTX), hypertonic saline/hydroxyethyl starch solution group (HSH) and hypertonic saline/hydroxyethyl starch solution combined with pentoxifylline group (HSH/PTX),12 rats in each group. The C group rats were only anesthetized, and the right femoral artery and femoral vein catheter inserted, but no shock, no resuscitation. The other groups'rats were bled to remain a mean arterial pressure of (35±5) mmHg for 1h. According to Wigger's method, the rats'models of severe hemorrhagic shock were reproduced. The NR group rats were not resuscitated. The NR group rats were sacrificed at 1h after shock, and specimens were taken immediately after death to be tested. The RL group rats were given 3 times the amount of blood loss of RL which was constantly infused within 20 minutes. The RL/PTX group rats were given the same treatment as the RL group, but PTX (25mg/kg) was integrated into RL. The HSH group rats received HSH solution (6ml/kg) which was constantly infused within 5 minutes. The HSH/PTX group rats were given the same treatment as the HSH group, but PTX (25mg/kg) was integrated into HSH. Normal saline (6ml/kg·h) was constantly infused for 4 hours after fluid resuscitation in four fluid resuscitation groups. The mean arterial pressure (MAP) of rats was monitored before shock, before resuscitation and 0.5h, 1h,2h,3h,4h after resuscitation, respectivly. At the end of the experiment, the rats were sacrificed to measure the concentration of i-FABP in plasma by ELISA. The levels of MPO, MDA and SOD in the intestinal tissue were detected by spectrophotometric method. The damage degree of intestinal tissue was observed with optical microscope examination. Epithelium, organelles connections, the structure and apoptosis in intestinal specimens was observed with transmission electronic microscopeResults 1.Oxidatie stress reaction:compared with group NR, LR, LRPTX, HSH, the pathological damage was lessened significantly, the levels of MPO, MDA in intestinal tissue and i-FABP in plasma were decreased, activity of SOD in intestinal tissue was increased significantly in group HSH/PTX (P<0.01).2. Optical microscope examination of intestinal tissue by hematoxylin-eosin (HE) staining, Chiu's damage score, electron ultrastructure pathology examination, i-FABP content in plasma:Compared with group NR, group RL, group RL/PTX and group HSH, Chiu's damage score and i-FABP content in plasma were reduced obviously in group HSH/PTX (P<0.01). The intestinal injury observed by optical microscope examination and electron ultrastructure pathology examination was milder in group HSH/PTX.Conclusion HSH/PTX might protect the intestinal mucous by increasing the antioxidant cell factor activity (SOD) and decreasing the levels of oxidant cell factors (MPO, MDA). |
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