| Objective: The study on intraperitoneal injection propofol in rats, which observes the influence of the survival area of island skin flap and histological varieties after ischemia-reperfusion(I/R), is to infirm the effects on preventing and curing the putrescence of flap, and to explore a new effective way for the enhance of skin flap transplantation survival rate. Trough the quantificational measurement of the concentration of nuclear factor-kappa B(NF-κB) and Intercellular Adhesion Molecule 1(ICAM-1) in skin flap, preliminarily discuss the mechanism of the propofol to protecting flap from ischemia-reperfusion injury, and provide reliable experimental basis for more extensivly applying to clinical application.Method:Select 80 healthy Wistar rats, male or female.The rats were divided into 4 groups randomly with each group 20 rats. P0 was control group, P50 was propofol 50mg/kg group, P100 was propofol 100mg/kg group, P150 was propofol 150mg/kg group. Before making animal model, inject distilled water to group P0 rat's abdominal cavity, inject propofol 50mg/kg to group P50 rat's abdominal cavity, inject propofol 100mg/kg to group P100 rat's abdominal cavity, inject propofol 150mg/kg to group P150 rat's abdominal cavity. Refer to Manson's method, make a island skin flap on rat's right lower quadrant with a size of 3.0cm×7.0cm, which blood supply was by abdominal wall superfic blood vessel. Obstruct the femoral artery before abdominal wall superfic blood vessel femoral artery. Take out of bulldog clamp and make the femoral artery recanalizate. Get specimen at I/R2h, I/R8h, I/R24h, detect skin flap tissue MDA and MPO by colorimetric method , detect skin flap tissue ICAM-1 and NF-κB by immunohistochemistry.Observe the survival situation of skin flap and calculate survival rate on the 7th day after operation. All data was collated and processed by SAS 8.0 software. One-way ANOVA was used to compare the difference between different group mean, least significant difference was used to compare the two groups if there was significant difference by one-way ANOVA analysis. Linear correlation was used to two analysis variable dependablity. P<0.05 was with a significant difference.Results1,The general observation:The 7th day after operation, for each groups, the color of the near section of flap was light red and with good elasticity, and a little abdominal hair grew up. For the middle and far section of the control group, the color was black and covered with scabs, and Less flexible. Little blood can be seen when lifting up the flap at the original position, much inflammatory exudates under the flap. Group P50: middle section: the color of the center flap gradually become darken. There was a little scab formation on surface, no hair grew, elasticity decrease. Far section: The color was black, and covered with scabs, bad elasticity. There can be seen a little bleeding when lift up the flap at the original position, a little inflammatory exudates under the flap. The survival situation of skin flap of Group P100 and P150 was proximate. For the middle section, the color of the center flap gradually becomes darken, there is no scab formation on surface, a little hair grew. For the far section, the color is black in peripheral area and covered with a little scabs,the elasticity was diminished and blood activity can be seen when lifting up the flap at the original position, and the quantity is large. There is no hematocele under the flap, little effusion, and vessels were abundance. The flap color and elasticity of combination group is better than the group that use the drug .According to the results of the observation, the survival situation of group P50,P100,P150 is better than group P0, and group P100 and P150 is obviously better than group P50.2,The ratio of surviving skin flap:Group P150(68.9±7.2)%﹥group P100(68.0±6.7)%﹥group P50(52.9±6.7)%﹥group P0(42.5±5.1)%. There was a significant difference between flap survival area of treatment group and the control group (F=78.52, P﹤0.01). Post hoc multiple comparisons, Group P150 had no significant difference compared with group P100(P﹥0.05), and among other groups, there were all significant differences(P﹤0.01).3,laboratory examination results3.1,The observation under light microscope:The 7th day after operation, in each group: Cuticle atrophy was not obvious or partly damaged, the skin layers of structure, no significant changes in neutrophil infiltration, freshmen capillary can be seen in cutis, subcutaneous fibrous tissue hyperplasia, fiber array regually. The control group: Cuticle and cutis were completely necrosis, the structure was disordered, a mount of neutrophils infiltration, muscles fiber obviously necrotized.3.2,MDA test results:The MDA value of the flap in different group was about 2.23 nmol / mgp before I/R and increased gradually with the time after I/R .The MDA value reached the peak value at I/R 24h .Compariing the MDA value of the four groups ,there were significant difference(F=137.65 ,P < 0. 01); Comparing MDA value with the control group respectively, the low/middle/high-dose group descended obviously and had significant difference(P < 0. 01); middle/high-dose group was obviously lower than low -dose group(P<0.05). There were no significant difference between high-dose group and middle-dose group(P >0. 05).3.3,MPO testing results:The MPO value of the flap in different group was about 0.41U/g before I/R and increased gradually with the time after I/R .The MPO value reached the peak value at I/R 24h .Comparing the MPO value of the four groups ,there were significant difference(F=137.65 ,P < 0. 01); Comparing MDA value with the control group respectively, the low/middle/high-dose group descended obviously and had significant difference ( P < 0. 01 ) ; middle/high-dose group was obviously lower than low -dose group(P<0.05). There were no significant difference between high-dose group and middle-dose group(P >0. 05) 3.4,The results of immunohistochemical staining of ICAM-1:The expression of positive ICAM-1 protein located the cytoplasm of the neutrophilic granulocytel and vascular endothelial cell. The expression of ICAM-1 was seldom at I/R 2h. The expression of ICAM-1 increased obviously at I/R 8h,.The expression of ICAM-1 increased obviously at I/R 24h. The peak value of the four groups had significant differences ( F= F=53.71,P<0.01); Comparing the expression of ICAM-1 with control group, the low/middle/high-dose group decreased obviously and had significant difference(P<0. 05); middle/high-dose group descented obviously than low-dose group(P<0.05), there were no significant difference between high-dose group and middle-dose group(P >0. 05).3.5,The results of immunohistochemical staining of NF-κB:The expression of positive NF-κB protein located the cytoplasm of the neutrophilic granulocytel and vascular endothelial cell and strong expression sometimes at nucleus. The peak value of the control group NF-κB protein positive expression appeared at I/R 2h, however the peak value of the treatment group appeared at I/R 8h. The peak value of the four groups had obvious significant differences ( F= F=38.26,P<0.01 ) ; Comparing the expression of NF-κB with control group, the low/middle/high-dose group decreased obviously and have significant difference ( P<0. 05 ) ; middle/high-dose group descented obviously than low-dose group(P<0.05), there were no significant difference between high-dose group and middle-dose group(P >0. 05); At I/R 8h, the expression decreased in the control group, but increased in the treatment group. At 24h I/R, the expression of NF-κB decreased obviously in the different groups and have no significant difference(P>0.05).3.6,Dependencies between NF-κB and ICAM-1:The expression of NF-κB and ICAM-1 was positive correlation, group P0 ( r=0.6066,P<0.01 ) , group P50 ( r=0.4876,P<0.05 ) , group P100(r=0.5632,P<0.05), group P150(r=0.5955,P<0.01. Conclusions:1,Activating of ICAM-1 and NF-κB may play a part in the pathological process of ischemical reperfusion injury of skin flap.2,Propofol may inhibit the espression and activating of NF-κB by cleaning up free radicle,cutting down inflammatory factor and inhibiting lipid peroxidation reaction of ischemic of skin flap,which can low the espression and activating of NF-κB and reduce the sticking gathering of heterophil granulocyte. So, Propofol may protect ischemical reperfusion injury of skin flap.3,The propofol optimal dosage is 100-150mg/Kg on rats intraperitoneal injection... |
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