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目的本实验旨在利用SAP大鼠动物模型检测胰腺组织中基质金属蛋白酶-2,9(MMP-2,9)的表达情况,以及血清中α2-巨球蛋白含量的变化。比较分析SAP中的α2-巨球蛋白和MMP-2,9的改变特征,进一步地分析各指标对SAP的影响及其作用,为SAP的防治提供理论依据。方法 (1)建立SAP大鼠动物模型,观察大鼠胰腺湿重、胰腺组织病理改变。(2)成模后采用免疫组化方法检测MMP-2,9的表达;采用ELISA法检测血清中α2-巨球蛋白含量的变化。结果 (1)胰腺光镜观查:SAP组肉眼可见胰腺坏死出血灶,光镜见间质肿胀,大量炎细胞浸润,红细胞渗出,腺泡细胞和脂肪坏死。对照组无明显异常变化。(2)ELISA法显示,实验组同对照组大鼠比较,α2-巨球蛋白含量升高。(3)免疫组化结果显示,SAP组MMP-2,9呈高表达,结果可见胰腺毛细血管内皮细胞胞浆及细胞外基质细小的棕黄染色颗粒。SAP组MMP-2,9表达阳性率为100%,其中强阳性有12例。而对照组表达阳性率为10%,其中弱阳性2例。2组间比较差异有显著性(P<0.01)。结论 (1)MMP-2、9在SAP中因炎性细胞受炎症因子的激活而大量释放,与SAP病情发生发展有密切关系;(2)α2-巨球蛋白在SAP中含量升高,对SAP有诊断和判定预后作用,从而成为SAP诊治的新靶点。
Objective This experiment aimed to detect the expression of MMP-2, 9 in pancreatic tissue and the changes of α2-macroglobulin in serum by using animal model of SAP. The changes of α2-macroglobulin and MMP-2, 9 in SAP were compared and analyzed. The influence of each index on SAP and the role of SAP were further analyzed to provide a theoretical basis for the prevention and treatment of SAP. Methods (1) SAP rat model was established to observe the pancreas weight and pathological changes in the pancreas. (2) The expression of MMP-2, 9 was detected by immunohistochemistry after the model was established. The changes of the content of α2-macroglobulin in serum were detected by ELISA. Results (1) Pancreatic light microscopy: Pancreatic necrosis hemorrhagic lesions were observed in macroscopic eyes of SAP group. Interstitial swelling was observed by light microscopy. A large number of inflammatory cell infiltration, erythrocyte effusion, acinar cells and fat necrosis were observed. The control group showed no abnormal changes. (2) ELISA showed that the experimental group compared with the control group rats, α2-macroglobulin increased. (3) The result of immunohistochemistry showed that the expression of MMP-2,9 in SAP group was high. The results showed that the cytoplasm of pancreatic capillary endothelial cells and small extracellular matrix of brown-yellow staining particles. The positive expression rate of MMP-2,9 in SAP group was 100%, of which 12 were strongly positive. The positive rate of the control group was 10%, of which 2 were weakly positive. The difference between the two groups was significant (P <0.01). Conclusions (1) The release of MMP-2 and MMP-9 in SAP due to the inflammatory cytokines released by inflammatory cytokines is closely related to the development of SAP. (2) The content of α2-macroglobulin in SAP is increased, SAP diagnosis and determine the prognosis, which has become a new target for SAP diagnosis and treatment.