ObjectiveTo explore the mechanism of liver capillary permeability in rats with severe acute pancreatitis (SAP). MethodsTotally 40 healthy Sprague-Dawley (SD) rats were randomly divided into two groups: sham operation (SO) group and SAP group, SAP group were divided into four subgroups according to sampling time (3 h， 6 h, 12 h， and 24 h). The model was established by injecting 5% sodium taurocholate retrogradely into pancreaticobiliary ducts. The changes of tumor necrosis factor-α (TNF-α), pathohistology, and tissue moisture content were compared among different groups. Liver occludin protein expression was analyzed by immunohistochemistry method, and occludin mRNA was measured by RT-PCR. ResultsThere was no significant pathological changes of liver tissue in the SO group. Typical pathological changes of SAP， such as interstitial edema, vasodilatation, infiltration of inflammatory cells, were found in the SAP group. TNF-α level and tissue moisture content of each phase increased gradually, and the highest level appeared at 24 h within the observing period. The two above indicators at different time point in subgroups were significantly different from each other and higher than those in the SO group (Plt;0.05)． In the SAP group, the expression of occludin and it’s mRNA began to decrease at 3 h to the bottom at 6 h and rebounced significantly at 12 h, 24 h compared with those at 6 h (Plt;0.05), but still lower than those in the SO group (Plt;0.05). ConclusionUpregulation in TNF-α and subsequent downregulation in occludin protein and mRNA maybe bly related to the severe liver capillary permeability in rats with SAP.
ObjectiveTo study the expression of zonula occluden-1 (ZO-1) in ileum tissues and the possible mechanism of intestinal mucosal barrier injury in rats with severe acute pancreatitis (SAP). MethodsFifty SD male rats were randomly divided into sham operation group and SAP group, then SAP group was divided into four subgroups with 10 rats in each subgroup according to the sampling time of 3, 6, 12, and 24 h. The SAP model was made by injecting 5% bovine sodium deoxycholate into biliarypancreatic duct with Aho’s method. The rats were killed at 3, 6, 12, and 24 h after making model. The rats in the sham operation group were killed directly. Tumor necrosis factor-α (TNF-α), diamine oxidase (DAO), and histological changes in pancreatic and intestinal pathologies were observed. At the same time, the ZO-1 protein and mRNA expressions of ileum tissues were detected by immunohistochemistry and RT-PCR, respectively. ResultsCompared with the sham operation group 〔TNF-α: (10.83±0.96) ng/L; DAO: (354.79±3.67) U/L; ZO-1 protein: (10.40±0.45) score; ZO-1 mRNA: 0.878±0.014 8〕, the levels of TNF-α at different time 〔3 h: (125.30±0.94) ng/L; 6 h: (181.89±4.93) ng/L; 12 h: (230.58±1.28) ng/L; 24 h: (198.89±4.83) ng/L〕 were significantly higher (Plt;0.05), the activities of DAO 〔3 h: (235.77±0.67) U/L; 6 h: (117.22±5.58) U/L; 12 h: (106.69±1.39) U/L; 24 h: (91.18±1.09) U/L〕 were significantly lower (Plt;0.05), ZO-1 protein 〔3 h: (8.70±0.22) score; 6 h: (3.73±0.19) score; 12 h: (3.92±0.22) score; 24 h: (4.29±0.30) score〕 and mRNA (3 h: 0.806±0.020 7; 6 h: 0.370±0.015 8; 12 h: 0.502±0.019 2; 24 h: 0.562±0.030 3) expressions of the ileum tissues were significantly lower (Plt;0.05) in the SAP group; Meanwhile, the necrosis of ileum mucous membrane chorioepithelium, angiorrhexis and hemorrhage, and inflammatory cell infiltration in the pancreatic and ileum tissues were also observed. ConclusionThe decrease of expression of ZO-1 in ileum tissues is one of the vital causes for mucosal barrier injury in SAP, probably through acts the excessive release of inflammatory cytokines TNF-α and the decrease of DAO activity.
The contents of lipid peroxides(LPO)and vitamin E(V.E)and some functional index and histologic changes in the lungs from the the rabbit models of acute cholangitis of severe type(ACST)were measured dynamically.The results revealed that the V.E content decreased strikingly from 6 hours and the LPO level increased progressivelg from 12 hours in the lungs.Simultanuosly,the congestion and neutrophil infiltreation in the lung mesenchyme,and the endothelial cell damage and thrombosis in the lung blood capillaries had been observed.These suggest that acute lung injury induced by ACST is referable to the lipid peroxidation damage to the lung blood capillaries which is due to increased LPO and decreased antioxidants including V.E.
Objective To investgate the expression of p38 mitogen-activated protein kinase （p38MAPK） in lung tissue of rats with severe acute pancreatitis （SAP）， and to explore the relationship between p38MAPK and pulmonary capillary barrier injury. Methods Forty male and healthy Sprague-Dawley （SD） rats were randomly （random number method） divided into sham operation （SO） group and SAP group， then rats of SAP group were sub-divided into 3， 6， 12， and 24 h group， each group enrolled 8 rats， respectively. SAP model rats were established by injecting 5% sodium taurocholate solution retrograde into the biliopancreatic duct. ELISA method was used to test the serum tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）， and pathological changes in lung and pancreas tissues were observed by HE staining. Immunohischemistry method was used to detect phosphorylated p38 （p-p38） protein and aquaporin 1 （AQP1） protein of lung tissues. The expression level of AQP1 mRNA was measured by quantitative real-time PCR. Results Hyperemia， edema， and inflammatory cell infiltration were observed in lung tissues， abundance of necrosis， part gland structure fuzzy or even disappear were observed in pancreas tissues of all 4 time point groups. Compared with SO group， levels of serum TNF-α and IL-1β were significantly higher in 4 time point groups （P＜0.05）. Lower expression level of p-p38 protein was detected in lung tissues of SO group， while in the early stage of SAP （SAP 3 h group）， the expression level of p-p38 protein significantly increased， which peaked in 6 h group and was still higher than SO group in 24 h group （P＜0.05）. Compared with SO group， the expression levels of AQP1 mRNA and protein were significantly lower in all 4 time point groups （P＜0.05）， which had negative correlation with the levels of serum TNF-α，IL-1β， and the expression level of p-p38 protein （r=-0.87， P＜0.05；r=-0.88， P＜0.05；r=-0.78， P＜0.05）. Conclusion The decrease of AQP1 protein in lung tissue is one of the vital causes for pulmonary capillary barrier injury in SAP， which probably works by the activation of p38MAPK and the excessive release of inflammatory cytokines.
Objective To establish a stable and reliable lung injury model caused by severe acute pancreatitis（SAP）in rats， which is helpful to study the acute lung injury （ALI）and acute respiratory distress syndrome （ARDS） induced by SAP.Methods Sixty Sprague-Dawley rats were randomized into ligature group （n=20）， traditional group （n=20），and sham operation group （n=20）. SAP model was established through retrograde injection of 5% taurocholic acid. After injection， the pancreatic duct of rats was ligated in ligature group， but not in traditional group. The lung damage and edema at 24 h after operaton and natural course of rats were observed.Results The ALI model of rats induced by SAP was established successfully in ligature group. The rats died of acute respiratory failure within 48 h in ligature group， the mortality was significantly higher than that in traditional group （100% vs.20%），P＜0.05. Pleural effusion occurred in four rats in ligature group， while no pleural effusion was found in rats in other two groups. The volume of ascites of rats in ligature group was （21.15±5.33） ml， which was more than that in traditional group 〔（7.75±2.66） ml〕，P＜0.05， while no ascites was found in rats in sham operation group. The level of serum amylase of rats in ligature group was （2 470.70±399.73） U/L，which was significantly higher than that in traditional group 〔（1 528.40±289.54） U/L〕 and sham operation group 〔（831.10±93.26） U/L〕，P＜0.001. The level of serum albumin of rats in ligature group was （6.90±1.66）g/L， which was significantly lower than that in traditional group 〔（13.10±0.99） g/L〕 and sham operation group 〔（16.20±0.92） g/L〕，P＜0.001.The lung wet-to-dry weight ratio （W/D） of rats in ligature group was 6.50±0.23， which was greater than that in traditional group （4.92±0.18） and sham operation group （4.61±0.16）， P＜0.001. The score of lung histopathologic of rats in ligature group was 29.25±1.07， which was significantly higher than that in traditional group （12.65±1.98） and sham operation group （0），P＜0.001. The score of pancreas histopathologic of rats in ligature group was 15.95±0.15，which was significantly higher than that in traditional group （13.75±0.66） and sham operation group （0.13±0.29），P＜0.001. Under transmission electron microscope， basement membrane of pulmonary capillary of rats in ligation group was destructive， the nuclei was dissolved， endothelial pinocytotic vesicles was functional active， and tight junctions between capillary endothelial cells were blurred and even ruptured. Moreover， tight junctions between alveolar epithelial cells were destructive. Pathological changes of lung ultrastructure of rats in ligation group were more severe than that in traditional group， while no pathological change of lung ultrastructure was observed in rats in sham operation group. Conclusions Injury process and pathogenesis of ALI or ARDS clinically caused by acute gallstone pancreatitis can be reproduced in this animal model， which is suitable to explore the related mechanisms of ALI caused by SAP and provides good animal model for the study of ALI caused by SAP.
The authors observed the progressive changes of the platelet activity in 25 cases of acute cholangitis severe type(ASCT)within 2 weeks of pre-and-post-operation.The results revealed that there are signficant changes of the platelet activity in ASCT.The levels of the platelet activity were proportional to the severity of disease and degree of biliary duct obstruction.Persistency of abnormality of platelet activity may predict the worse of disease and bad prognosis.The results are useful to understand the complex changes of pathophysiology in ACST,to the mechanism of multiple organic failure.
ObjectiveTo investigate the possible mechanism of cucurmosin on apoptosis in human pancreatic cancer cell line SW1990 in vitro. MethodsThe inhibition of cucurmosin on SW1990 cell was detected by MTT assay, the apoptosis was observed by transmission electron microscope, the apoptosis rate was analyzed by flow cytometry, and the protein level of caspase3 was determined by Western blot. ResultsAfter exposure to cucurmosin at 1.25, 2.50, 5.00, 10.00, 20.00, 40.00, and 80.00 μg/ml for 24, 48, and 72 h, the proliferation of SW1990 cell was inhibited in a time-and dose-dependent manner (Plt;0.05). At 72 h after 40.00 μg/ml cucurmosin treatment, the typical apoptosis changes and apoptotic bodies were observed by transmission electron microscope. After exposure to cucurmosin at 0, 2.50, 10.00, and 40.00 μg/ml for 72 h, the apoptosis rate increased gradually as (0.30±0.11)%, (18.93±1.06)%, (28.00±2.07)%, and (49.93±3.25)%, respectively (Plt;0.05). The expression of caspase-3 protein was elevated gradually (Plt;0.05). ConclusionCucurmosin may induce the apoptosis of pancreatic cancer cell through up-regulating the expression of caspase-3.