| Objective:Study acute liver injury rat serum cytokine TNF alpha, IL-6 and HSP70 expression within the liver tissue changes, so as to clinical research early diagnosis in the treatment of acute liver injury to provide a new thinking and theoretical basis. Methods: 40 healthy adult male Spague-Dawlay (SD) rats were divided into four groups randomly after adaptive feeding one week, sham operation group (A) 10,50% liver excision group(B) 10,70% liver excision group(C) 10,80% liver excision group(D)10. Fasting 12 h, free access to water, to ketamine,100 mg/kg by intraperitoneal injection anesthesia, under sterile condition upper abdominal transverse incision, According to Kubota T method improvement, modified with no blood intraoperative model cut liver. After laparotomy dissociating liver surrounding tissues,exposure hepatic artery and portal vein and biliary. With nearly portal ligation method excision lobe of liver. After checking no bleeding closed the abdominal cavity. Refer to ShiJiHua literature method building model. According to pre-test results, after 24 hours, both control group and model group were killed,Blood was collected through Automatic biochemical analyzer to determinated rat's blood lipid and serum aminopherase in each group inculded TB,TBA,ALT,AST. Enzyme-linked immunosorbent adsorption method testing serum IL-6, TNF-a. Model is successful, through Sodium pentobarbital anesthesia, Then the liver was taken out rapidly to take it's wet weight, the liver index was calculated as follows:liver wet weight/body mass×100%. And in the middle of the left lobe of liver was cut into 1cm×1cm×1cm as samples, placed in 10% formalin fixed and paraffin sections. The liver tissue samples were taken to preparate paraffin sections. The liver tissue pathological changes were observed through the hematoxylin- eosin (HE) staining under light microscope and the degree of liver fibrosis was detected. The expression of protein of HSP70 in liver tissue were examined by immunohistochemistry,by image analysis system, integrated optical density was analyzed. The gray value was analyzed by Biod-rad softwares. Optical density value denotes protein expression amount of HSP70. Measuring data was demonstrated with mean±standard deviation (x±S).With t test, the groups were compared with variance analysis. If there was homogeneity of variance, one-way ANOVA analysis was used,and if there was heterogeneity of variance H test was used. The difference between the model group and the control group in the serum, pathology and extent of apoptosis was compared. Count data was used chi-square test; the difference of P<0.05 has statistical significance. Results:All 40 rats were entered into the final analysis.(1) General condition each rat postoperative day 1 survival 100%. The control group animals is in high spirit,flexible and active, eating water normal and fur glossiness is good.The liver function index of postoperative rats increased significantly. Model rats, appear different degree of spiritual sluggish and activity decreased.90% liver resection model group postoperatie signs of liver failure, performance for postoperative prolonged, sleepiness and awaken to stimuli as weak, eyes appear hemorrhagic secretions and rat hair loss, abdominal incision ooze blood, skin is yellow, niaoshao deep yellow color, etc. (2) HE stain reveals, A group of liver tissue without obvious anomalous change; Liver excision group rat liver tissue shows extensive liver cell vacuoles, cells swell, sample degeneration of the bubble plasma cells occur within the nucleus, vacuoles of varying sizes. Liver sinus apparent expansion, scattered liver cell pointlike necrosis, collect abbacy boundaries clear, a few inflammatory cells infiltrating, lobular morphology complete. Group D visible liver cell cytoplasm and small vacuoles within merged into big vacuoles central, suspension, the nucleus, the cells form blank cytoplasm significantly enlargement. (3) The immunity histochemistry staining method showed that the expressions of Pgp3 in liver were dyed yellowish brown under the high power objective. With integral optical density (IOD) for analysis, the difference between the four groups was statistically significant. (4) ELISA test rat serum cytokine IL-6, TNF alpha display made each model group after mould 24h rat serum IL-6, TNF alpha level is significantly higher control group (p<0.01), model group is compared between each group have significant difference (p< 0.01, p<0.05). Conclusion:(1) Successfully established a rat acute liver injury model, this model is mainly for the liver function index increased significantly, appear different degrees of liver damage, the liver empty bubble sample degeneration obvious, even happen plaque-forming necrosis. (2) Acute liver injury rats' serum TNF alpha, IL-6 levels significantly higher and TNF alpha, IL-6 can reflect the liver tissue damage degree. (3) Acute liver injury in rats, liver tissue HSP70 express obvious enhancement; And HSP70 expression liver damage the heavier the more obvious.
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