The Gastric Mucosa Reperfusion Injury Induced After CO₂ Pneumoperitoneum And The Protective Effect Of Propofol In Rats

ObjectiveObserving the gastric mucosal ischemia-reperfusion injury induced after CO₂ pneumoperitoneum and the protective effect of propofol in rats.Material and Methods1.experiment materialsAdult male Wistar rats(by the China Medical University Hospital Shengjing Department of Laboratory Animals)about 280～320 grams,fasting 24 hours before the experiment,free drinking water.Experimental one week prior to the adaptability of feeding.2.groupingThey were randomly divided into four groups,each with six.GroupA:control group,the tail vein injection pump saline(10 ml/kg/h)1h.GroupB:ischemia group,in Group A on the basis of 20 mmHg for 1h pressure pneumoperitoneum.GroupC:ischemia-reperfusion group,with the disposal of Group B,1h after pneumoperitoneum,and maintain a 0.5 h-abdominal pressure.GroupD:Protection Unit,20 mmHg pneumoperitoneum 1h,without pneumoperitoneum 0.5h,the entire process of the tail vein injection pump 1g/1 of propofol 10ml/kg/h(propofol with saline liquid to 10%).3.methods 10%of chloral hydrate aldehyde(3 ml/kg body weight)intraperitoneal injection of anesthesia.Then fixed on the operating table animals,the tail vein puncture,for infusion of propofol or saline.Abdominal alcohol disinfection,the umbilical will pneumoperitoneum needle inserted into the abdominal cavity.A group not to charge into the abdominal CO2,maintaining 0-abdominal pressure 1h;B,C,D groups filling the CO2,gas and abdominal pressure at 20 mmHg.A,B,C group saline injection pump 10 ml/kg/h;D Group infusion of propofol 10 mg/kg/h,propofol solution(10 mg/ ml)with saline to 10%.C,D Group pneumoperitoneum 1h after the cessation of pneumoperitoneum,0-abdominal pressure to maintain 0.5h,each type of liquid injection pump and time,gas and abdominal pressure and reperfusion time table 1.4.indicators of observation and biochemical index(1)Light microscopy of gastric mucosa:At the end of the experiment,open,free full stomach rapidly along the greater curvature of the ripped,cold saline rinse contents, in the central part of the proventriculus lesions clear position from 2×3mm gastric 10 24-48 h%formalin-fixed,the light microscope specimens to be done,conventional embedded in paraffin,slicing line 4 um,HE staining.(2)Electric microscopy of gastric mucosa:Gastric mucosa specimens obtained the same way as(1),1×0.5mm from gastric mucosa at 2.5%glutaraldehyde in double distilled water fixative,4℃refrigerator preservation,electron microscopy specimens to be done.(3)Superoxide dismutase SOD::Gastric mucosa specimens were placed in the rapidly frozen in liquid nitrogen reserve.After reunification weighing 4℃normal saline,10%of homogenate,4℃,100000 r/min for 15 min,the supernatant from -80℃preservation.and determined by the way of xanthine oxidase enzymatic assay. Through xanthine and xanthine oxidase reaction system to produce Superoxide radical (O.^-),the latter form of hydroxylamine nitrite in the reagent showed the effects of purple,with color of its absorbance.(4)Malondialdehyde(MDA):Preparation of specimens Ditto.By thiobarbituric acid colorimetric assay.Use of lipid peroxidation main degradation product malondialdehyde in the acidic conditions of the two thiobarbituric condensation to form a red substance in 532 nm colorimetric assay.(5)Nitric oxide(NO):Preparation of specimens Ditto.Use nitrate reductase method.NO chemical properties and lively,and in vivo metabolism quickly to NO₂^-and NO₃^-,and NO₂^- further into NO₃^-,using nitrate reductase NO₃^- specificity will be reduced to NO₂^- by the color of their deep level of concentration.(6)Glutathione peroxidase(GSH-PX):GSH-PX could promote hydrogen peroxide and glutathione reaction of water and oxidized glutathione,GSH-PX,the vitality of its available enzymatic reaction to the speed that this enzymatic reaction of glutathione depletion can be obtained enzyme activity.(According to various indicators of detection reagents and instructions for use).5,statistical analysisSPSS13.0 software system used to conduct statistical processing,measurement data to mean±standard deviation said.Group and the group using t-test analysis.It has statistical difference when p＜0.05.Result1.light microscopy findingsGroupA:normal gastric mucosa.GroupB:submucosal small amount of inflammatory cell infiltration,some mild gastric mucosal surface erosion.GroupC:gastric mucosa depression,a shallow ulcers,submucosal infiltration of inflammatory cells.GroupD:reduce the infiltration of inflammatory cells,relatively smooth surface of gastric mucosa close to normal gastric mucosa.2.electron microscopy findingsGroupA:parietal cells:There is a great deal of micro-bubble pipe system,and normal mitochondria in parietal cells.The nuclear mumbrance is smooth. adelomorphous cell:There are large number of rough endoplasmic reticulum and some normal structure of mitochondria.GroupB:parietal cells:a small number of mitochondria are floc change in cytoplasm, irregular nuclei,micro-bubble pipe system to be reduced. adelomorphous cell:The irregular-shaped nucleus,rough endoplasmic reticulum expansion and mitochondrial swelling,decreased density matrix, fuzzy appearance,with flocculent of change.GroupC:irregular-shaped nuclear,heterochromatin-set,the micro-bubble system decline seen myelin-like structure in the cytoplasm,mitochondrial membrane damage,membrane swelling. adelomorphous cell:nuclear membrane swelling,heterochromatin-set, rough endoplasmic reticulum excessive expansion.GroupD:parietal cells:micro-bubble system started increasing. adelomorphous cell:rough endoplasmic reticulum relative ease swelling.3,biochemical indicatorsIschemia-reperfusion group increased MDA,SOD,NO,GSH-PX lower,compared with the other groups were statistically significant.Propofol protection group and ischemia-reperfusion group compared to MDA decreased SOD,NO,GSH-PX rise between the two groups,the difference was significant.Conclusion1.Carbon dioxide insuffiation on rat gastric mucosa can cause ischemia -reperfusion injury.2.Propofol on rat gastric mucosa in ischemia-reperfusion injury protection.