Effect Of Expression Of CD50 In Macrophage On SIRS Induced By SAP With Combined Treatment Of Traditional Chinese And Western Medicine

Background and objective: Acute pancreatitis (AP) is a common surgical disease, whose clinical pathological change is complex, with various responses to treatment. Light edematous pancreatitis is a self-limiting disease with fine responses to treatment and prognosis, while severe acute pancreatitis (SAP) is disease with acute onset and rapid progress, which can lead systemic inflammatory response syndrome (SIRS)at early stage. Although there is rapid development in the treatment of SAP, the mortality rate is as high as 10 ~ 30%. SIRS is considered the main reason leading to death. A large number of studies have confirmed that the phagocytosis ability of macrophage in clearing apoptotic neutrophil decrease in SIRS, resulting in the release of a great number of inflammatory mediators increased systemic inflammatory response. CD50 molecule on the cell membrane of macrophages plays an important regulatory role in the clearance of apoptosis neutrophil, laying the foundation for macrophage identifying, removing apoptosis neutrophils. Therefore, the study of macrophage expression of CD50 in SIRS induced by SAP and its expression in different treatment is significant for treating SAP-induced SIRS and reducing the mortality rate. This experiment is to study the effect of traditional Chinese medicine Yinchen Chengqi Decoction, Dexamethasone and combination of the two drugs on expression CD50 protein in macrophage in SIRS induced by SAP rat model.Methods: SD rat SAP / SIRS model [3] was produced by injecting 1.5% sodium deoxycholate retrogradly into pancreatic duct through duodenal papilla. 75 SD rats were randomly divided into sham operation group (SO), severe acute pancreatitis SIRS model group(SIRS/SAP),Yinchen Chengqitang Decoction treatment group(YCCQT), Dexamethasone treatment group(DEX), traditional Chinese associates with western medicine treatment group (ZXY), dose of the Yinchen Chengqi Decoction was 1ml/100g (ig),while dose of Dexamethasone was 1mg/100g (intraperitoneal injection), the treatment group in two hours and after producing the model with a re-delivery in 12 hours. Model samples were collected after 24h for detection.Pathological changes and pathology score of pancreat were observed with microscope in each group; the positive rate of CD50 in macrophages was detected by flow cytometry; Use western blotting to detect differences of macrophage CD50 protein expression level between each group; the differences of levels of CD50 protein were detected by immunofluorescence.Results:1.Pathological changes of pancreas in each group: (1)compared with SO group, pathological changes in SI Group were significantly obvious (edema: 3.50±0.21 vs 0.290±0.05, inflammation: 2.90±0.15 vs 0, hemorrhage: 2.10±0.18 vs 0, necrosis: 2.80±0.24 vs 0, P <0.01); (2)compared with the SI group,various improvement in treatment groups can be seen with statistical significance (P <0.05), particularly in ZX group (edema: 3.00±0.14 vs 3.50±0.21, P <0.05, inflammation: 1.40±0.23 vs 2.90±0.15, hemorrhage: 0.40±0.15 vs 2.10±0.18, necrosis: 1.40±0.12 vs 2.80±0.24, P <0.01).2.Expression of macrophage CD50 positive rate in Flow cytometry:(1)Compared with SO group, CD50-positive macrophages in SI group was significantly decreased(12.32±8.82% vs 36.52±12.61%, P <0.01);(2) Compared with SI group, macrophages CD50-positive rate slightly increased in YC Group (12.47±4.36% vs 12.32±8.82%, P <0.05), while DE Group(26.98±3.72% vs 12.32±8.82%, P <0.05) and ZX group(35.35±5.74% vs 12.32±8.82%, P <0.01) were significantly increased;(3)Compared within treatment groups: the CD50-positive rate was significantly increased in ZX group than inYC group and DE group (35.35±5.74% vs 12.47±4.36% vs 26.98±3.72%, P <0.05).3.Western Blotting Detection of macrophage CD50 protein expression:(1)Compared with SO group, SI group macrophage expression of CD50 were significantly different (0.47±0.09 vs 0.76±0.15, P <0.01);(2)Compared with the SI group: YC group macrophage CD50 expression levels were significantly different (0.47±0.09 vs 0.55±0.08, P <0.05), DE group macrophage CD50 expression levels were significantly different (0.47±0.09 vs 0.56±0.11, P <0.05), while the ZX group macrophage expression of CD50 were significantly different (0.47±0.09 vs 0.71±0.13, P <0.01);(3)To compare within treatment groups: macrophage CD50 expression levels in ZX group were significantly different (0.71±0.13 vs 0.56±0.11,0.71±0.13 vs 0.55±0.08, P <0.05 )comparing with DE groups and YC group; DE group compared with the YC group, macrophage expression of CD50 significantly different (0.56±0.11 vs 0.55±0.08, P<0.05).4.AU value of macrophage CD50 protein expression detected by Immuno- fluorescence:(1)Compared with SO group, expression of macrophage CD50 in SI group were significantly different (235.44±21.35 vs 354.72±28.46, P <0.01);(2)Compared with SI group: macrophage CD50 expression in DE group were significantly different (235.44±21.35 vs 275.43±30.27, P <0.05), macrophage CD50 expression in YC group are different (235.44±21.35 vs 267.39±25.36, P <0.05), while the macrophage expression of CD50 in ZX group were significantly different (235.44±21.35 vs 305.52±31.14, P <0.01);(3)Compared within treatment groups: macrophage CD50 expression in ZX group were significantly higher than YC group and DE group (305.52±31.14 vs 267.39±25.36,305.52±31.14 vs 275.43±30.27, P <0.05 ); compared with the YC group, macrophage CD50 expression in DE group were higher (275.43±30.27 vs 267.39±25.36, P <0.05).Conclusions:Compared with SO group, SI group has more serious pathological changes in the pancreas.The treatment group have varying degrees of improvement, with the most obvious effect in ZX group, implying that Yinchen Chengqi Decoction alone or Dexamethasone alone has less significant effect on pathological changes of rat pancreas than joint Application of Western and Chinese Medicine. Expression of macrophage CD50 in SI Group significantly decreased, the treatment groups increased in varying degrees, in which Chinese and Western Medicine has better effect on expression of macrophage CD50 protein than the single treatment group, suggesting that the SAP / SIRS model of the membrane of macrophages may be changed so that the CD50 in the surface of macrophage decreased significantly, affecting the apoptosis of macrophages and neutrophils adhesion and cell-to-cell signal transduction, resulting in decreasing phagocytic function of macrophages removing apoptosis neutrophil , and further rise to a large number of neutrophils accumulated in the body to release excessive inflammatory response to increase inflammatory mediators, so that a deterioration in conditions. Therefore, traditional Chinese and western medicine may be effective way to increase the capacity of macrophage phagocytosis and become treatment of SAP caused by SIRS.