前B细胞克隆增强因子（PBEF）在急性胰腺炎肺损伤大鼠肺组织中的表达与功能及清胰汤干预的实验研究
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摘要
目的
本实验将通过观察前B细胞克隆增强因子(Pre-B-cell colony-enhancing factor, PBEF)在重症急性胰腺炎(severe acute pancreatitis, SAP)肺损伤(acute lung injury, ALI)模型大鼠肺组织中的表达及功能,并观察清胰汤对SAP大鼠肺脏PBEF表达的影响。从新的角度揭示重症急性胰腺炎相关肺损伤的发病机制,同时探讨清胰汤的治疗作用机理,为临床上中西医结合治疗重症急性胰腺炎提供进一步理论基础。
方法
健康Spraghe-Dawley(SD)大鼠144只,体重200-220g,雄性,随机分为6组(SAP模型组,假手术组,地塞米松组,铃兰欣组,善宁组,清胰汤治疗组)每组24只,每组随机分为造模6h,12h,24h三个时相典亚组,各时相点亚组8只。SAP组和药物干预组开腹后用无损伤动脉夹夹闭胰胆管入肝门处,逆行胰胆管缓慢注入1.5%的去氧胆酸钠溶液(1ml/kg),取出动脉夹,关腹,建立SAP时ALI模型。地塞米松组于造模后立即静脉注射一次及造模后12h再次静脉注射一次,剂量：1 ml/kg。善宁组于造模后立即静脉注射一次及造模后12h再次静脉注射一次,剂量20μg/kg。铃兰欣组于造模后立即静脉注射一次及造模后1 2h再次静脉注射一次,剂量2mg/kg。清胰汤组造模后胃管注入中药煎液及造模后12h再次灌胃一次,剂量：10ml/kg。假手术组常规剖腹后翻动胰腺,关腹。分别于造模后6h、12h、24h处死相应时相点大鼠,取肺组织及动静脉血做病理检查和各项指标检测。
采用双抗体夹心(enzyme linked immunosorbent assay, ELISA)法测定大鼠血清中PBEF含量,免疫组化检测其在肺组织中的表达及组织定位；RT-PCR法检测肺组织中PBEFmRNA表达；Western Blotting法检测肺组织中PBEF蛋白含量；鲎试剂偶氮基质显色法定量检测血清内毒素含量；放免法检测TNF-a, IL-1 p,IL-8在肺组织中的表达；真空干燥法测定肺湿干重(W／D)比值；全自动生化分析仪检测动脉血气；酶法测血清淀粉酶(AMY),大体及镜下观察肺脏病理学改变。
结果
1.模型组血清PBEF含量升高、PBEFmRNA及其蛋白出现高表达,TNF-α、IL-1β和IL-8均明显高于假手术组。同时,模型组病理学表现为明显的肺损伤,肺W／D比值、血中内毒素、血中AMY等指标均较假手术组显著升高(p<0.01)。清胰汤、地塞米松、善宁可以下调血清PBEF、PBEFmRNA及其蛋白表达,减轻肺损伤程度,而铃兰欣作用不明显。
2.清胰汤组血中内毒素、TNF-α、IL-1 p、IL-8等指标下降显著(p<0.05)。善宁组抑制血中AMY作用最强((p<0.05)。地塞米松组对改善肺W／D比值、PaCO2、PaO2作用明显。
结论
1.应用1.5%的去氧胆酸钠逆行胰胆管内注射,大鼠的胰腺充血水肿明显,血清AMY、肺W/D、PaCO2的指标明显升高,随着时间的推移逐渐增高,较相同时相点假手术组显著增高Pa02呈现相反变化。还可见肺组织水肿、充血、炎细胞浸润,表明重症急性胰腺炎合并肺损伤的模型复制成功。
2.重症急性胰腺炎相关肺损伤时,内毒素水平升高,PBEF表达过度上调,进而启动TNF-α、IL-1 p和IL-8等炎性细胞因子的大量表达,在SAP相关ALl发病中起着十分重要的作用。
3.中药清胰汤通过下调PBEF的表达,减缓肺泡巨噬细胞的过度激活,减少TNF-α、IL-1 p和IL-8炎性细胞因子的大量表达,减少中性粒细胞(polymorphonuclear neutrophil, PMN)聚集,降低血中高水平的内毒素和AMY浓度、降低肺泡通透性等多个方面机制对肺组织起到保护作用。
4.善宁、地塞米松可以不同程度下调PBEF的表达,通过不同途径对不同指标产生影响,从而实现对ALl某种程度的保护作用。
5.中、西药物在SAP相关ALl发病的不同环节中,起着十分重要的作用,各有所长,中西医结合理论指导下的中西药联合应用,将会为临床SAP相关ALl的防治提供新的有效途径。
Objective:To observe the expression and function of pre-B cell cloning factor (PBEF) of the rat's blood and lung tissue in severe acute pancreatitis (SAP) associated acute lung injury (ALI) model, and the effects of Qingyitang on PBEF expression. To reveal a new perspective pathogenesis of severe acute pancreatitis associated with lung injury.Chinese medicine Qingyitang is used and the results may give us some new ideas for severe acute pancreatitis treatment.
Methods:One hundred and fortyfour health Spraghe-Dawley (SD) rats weight (200-220) g, male, were randomly divided into 6 groups (SAP model group, Sham operation group, and Dexamethasone group, Linglanxin group, Sandostatin group, Qing Yitang group). These 24 animals in each group were randomly divided into Oh,6h,12h,24h subgroup,8 rats were at each subgroup. All modles were induced by retrograde infusion of 1.5% sodium deoxycholate (1 ml/kg) into biliopancreatic duct of Spraghe-Dawley rats except sham operation group. Dexamethasone group, Sandostatin group, Linglanxin group were done with intravenous injection immediately after modeling the first and 12h once again, respectively intravenous dose: 1 ml/kg,20μg/kg,2mg/kg. Qingyitang was given in Qingyitang group with gastric canal injection immediately after modeling the first and 12h in dose:10ml/kg. In Sham group, the pancreas was only flipped after laparotomy. All the rats were sacrificed at 6 h,12 h,24 h respectively after operation, the lung tissue and arterial and venous blood were taken off for pathological examination and some indexes measurement.
The level of PBEF in lung was measured by enzyme linked immunosorbent assay(ELISA),the expression and location of PBEF were measured by immunohistochemistry. RT-PCR technique was used to detect the expression of mRNA levels of PBEF and Western Blotting was used to detect the expression of protein levels of PBEF, respectively. Quantitative chromogenic tachypleus amebocyte was used to detect the endotoxin. TNF-α, IL-1βand IL-8 were measured by radioimmunoassay respectively. Wet/dry weight ratio of lung were calculated. Blood gas analysis and serum amylase (AMY) was determined by biochemical method. Histopathological changes in lung were observed under light microscope.
Results:The expression of PBEF mRNA and protein in lung tissue, the levels of serum PBEF, TNF-α, IL-1β, IL-8, endotoxin,amyIase, lung W/D ratio, PaCO2 in arterial blood of model group were significantly higher in the same point than those of sham operated control group, meanwhile PaO2 in arterial blood decreased significantly (p<0.01). Some other indexes of ALI showed obvious pathogenic changes in model group and so did the pathogenic examination of lung tissue. Qingyitang Dexamethasone, Santonstadine could decrease the levels of serum PBEF, the expression of PBEF mRNA and its protein in lung tissue, eliminate the extent of lung injury, but Linglanxin could not do that well.
The decline of endotoxin, TNF-α, IL-1β, IL-8 in Qingyitang group was more significant(p<0.05). Santonstadine could reduce amylase content obviously. And Dexamethasone showed superiority in ameliorating lung W/D ratio、PaCO2 and PaO2.
Conclusions:
Infusion of 1.5% sodium deoxycholate into biliopancreatic duct of Spraghe-Dawley rats, the pathologic changes of lung were edema and congestive with large of neutrial cells infiltration and focal atelectasis, the levels of serum AMY, PaCO2 and W/D increased obviously, meanwhile PaO2 decreased significantly. All indexes showed that the APALI model is welldone.
During acute lung injury induced by severe acute pancreatitis, many factors have very important effects on pathogenisis of ALI, such as elevated level of endotoxin,overexpression of PBEF which activated overexpression of TNF-a, IL-1β, IL-8.
Qingyitang could exert protecting effects through decreasing PBEF expression,reducing the excessive activation of alveolar macrophages, expression of TNF-a, IL-1βand IL-8 in a large number of inflammatory factors, aggregation of PMN, the level of serum endotoxin and amylase and decreasing alveolar permeability and other aspects of mechanism.
Dexamethasone and Santonstadine could play their available role in ALI by eliminating PBEF of different levels. Both of them could impact different indicators through different ways.
The effects of Chinese and Western medicines on SAP were related to different aspects of the pathogenesis of ALI, and play an important role by their own strengths. Combining Chinese and Western medicine under the guidance of the integration of traditional and western medicine theory, will provide a new effective way to prevent and cure SAP and SAP associated ALI in clinic.
本实验将通过观察前B细胞克隆增强因子(Pre-B-cell colony-enhancing factor, PBEF)在重症急性胰腺炎(severe acute pancreatitis, SAP)肺损伤(acute lung injury, ALI)模型大鼠肺组织中的表达及功能,并观察清胰汤对SAP大鼠肺脏PBEF表达的影响。从新的角度揭示重症急性胰腺炎相关肺损伤的发病机制,同时探讨清胰汤的治疗作用机理,为临床上中西医结合治疗重症急性胰腺炎提供进一步理论基础。
方法
健康Spraghe-Dawley(SD)大鼠144只,体重200-220g,雄性,随机分为6组(SAP模型组,假手术组,地塞米松组,铃兰欣组,善宁组,清胰汤治疗组)每组24只,每组随机分为造模6h,12h,24h三个时相典亚组,各时相点亚组8只。SAP组和药物干预组开腹后用无损伤动脉夹夹闭胰胆管入肝门处,逆行胰胆管缓慢注入1.5%的去氧胆酸钠溶液(1ml/kg),取出动脉夹,关腹,建立SAP时ALI模型。地塞米松组于造模后立即静脉注射一次及造模后12h再次静脉注射一次,剂量：1 ml/kg。善宁组于造模后立即静脉注射一次及造模后12h再次静脉注射一次,剂量20μg/kg。铃兰欣组于造模后立即静脉注射一次及造模后1 2h再次静脉注射一次,剂量2mg/kg。清胰汤组造模后胃管注入中药煎液及造模后12h再次灌胃一次,剂量：10ml/kg。假手术组常规剖腹后翻动胰腺,关腹。分别于造模后6h、12h、24h处死相应时相点大鼠,取肺组织及动静脉血做病理检查和各项指标检测。
采用双抗体夹心(enzyme linked immunosorbent assay, ELISA)法测定大鼠血清中PBEF含量,免疫组化检测其在肺组织中的表达及组织定位；RT-PCR法检测肺组织中PBEFmRNA表达；Western Blotting法检测肺组织中PBEF蛋白含量；鲎试剂偶氮基质显色法定量检测血清内毒素含量；放免法检测TNF-a, IL-1 p,IL-8在肺组织中的表达；真空干燥法测定肺湿干重(W／D)比值；全自动生化分析仪检测动脉血气；酶法测血清淀粉酶(AMY),大体及镜下观察肺脏病理学改变。
结果
1.模型组血清PBEF含量升高、PBEFmRNA及其蛋白出现高表达,TNF-α、IL-1β和IL-8均明显高于假手术组。同时,模型组病理学表现为明显的肺损伤,肺W／D比值、血中内毒素、血中AMY等指标均较假手术组显著升高(p<0.01)。清胰汤、地塞米松、善宁可以下调血清PBEF、PBEFmRNA及其蛋白表达,减轻肺损伤程度,而铃兰欣作用不明显。
2.清胰汤组血中内毒素、TNF-α、IL-1 p、IL-8等指标下降显著(p<0.05)。善宁组抑制血中AMY作用最强((p<0.05)。地塞米松组对改善肺W／D比值、PaCO2、PaO2作用明显。
结论
1.应用1.5%的去氧胆酸钠逆行胰胆管内注射,大鼠的胰腺充血水肿明显,血清AMY、肺W/D、PaCO2的指标明显升高,随着时间的推移逐渐增高,较相同时相点假手术组显著增高Pa02呈现相反变化。还可见肺组织水肿、充血、炎细胞浸润,表明重症急性胰腺炎合并肺损伤的模型复制成功。
2.重症急性胰腺炎相关肺损伤时,内毒素水平升高,PBEF表达过度上调,进而启动TNF-α、IL-1 p和IL-8等炎性细胞因子的大量表达,在SAP相关ALl发病中起着十分重要的作用。
3.中药清胰汤通过下调PBEF的表达,减缓肺泡巨噬细胞的过度激活,减少TNF-α、IL-1 p和IL-8炎性细胞因子的大量表达,减少中性粒细胞(polymorphonuclear neutrophil, PMN)聚集,降低血中高水平的内毒素和AMY浓度、降低肺泡通透性等多个方面机制对肺组织起到保护作用。
4.善宁、地塞米松可以不同程度下调PBEF的表达,通过不同途径对不同指标产生影响,从而实现对ALl某种程度的保护作用。
5.中、西药物在SAP相关ALl发病的不同环节中,起着十分重要的作用,各有所长,中西医结合理论指导下的中西药联合应用,将会为临床SAP相关ALl的防治提供新的有效途径。
Objective:To observe the expression and function of pre-B cell cloning factor (PBEF) of the rat's blood and lung tissue in severe acute pancreatitis (SAP) associated acute lung injury (ALI) model, and the effects of Qingyitang on PBEF expression. To reveal a new perspective pathogenesis of severe acute pancreatitis associated with lung injury.Chinese medicine Qingyitang is used and the results may give us some new ideas for severe acute pancreatitis treatment.
Methods:One hundred and fortyfour health Spraghe-Dawley (SD) rats weight (200-220) g, male, were randomly divided into 6 groups (SAP model group, Sham operation group, and Dexamethasone group, Linglanxin group, Sandostatin group, Qing Yitang group). These 24 animals in each group were randomly divided into Oh,6h,12h,24h subgroup,8 rats were at each subgroup. All modles were induced by retrograde infusion of 1.5% sodium deoxycholate (1 ml/kg) into biliopancreatic duct of Spraghe-Dawley rats except sham operation group. Dexamethasone group, Sandostatin group, Linglanxin group were done with intravenous injection immediately after modeling the first and 12h once again, respectively intravenous dose: 1 ml/kg,20μg/kg,2mg/kg. Qingyitang was given in Qingyitang group with gastric canal injection immediately after modeling the first and 12h in dose:10ml/kg. In Sham group, the pancreas was only flipped after laparotomy. All the rats were sacrificed at 6 h,12 h,24 h respectively after operation, the lung tissue and arterial and venous blood were taken off for pathological examination and some indexes measurement.
The level of PBEF in lung was measured by enzyme linked immunosorbent assay(ELISA),the expression and location of PBEF were measured by immunohistochemistry. RT-PCR technique was used to detect the expression of mRNA levels of PBEF and Western Blotting was used to detect the expression of protein levels of PBEF, respectively. Quantitative chromogenic tachypleus amebocyte was used to detect the endotoxin. TNF-α, IL-1βand IL-8 were measured by radioimmunoassay respectively. Wet/dry weight ratio of lung were calculated. Blood gas analysis and serum amylase (AMY) was determined by biochemical method. Histopathological changes in lung were observed under light microscope.
Results:The expression of PBEF mRNA and protein in lung tissue, the levels of serum PBEF, TNF-α, IL-1β, IL-8, endotoxin,amyIase, lung W/D ratio, PaCO2 in arterial blood of model group were significantly higher in the same point than those of sham operated control group, meanwhile PaO2 in arterial blood decreased significantly (p<0.01). Some other indexes of ALI showed obvious pathogenic changes in model group and so did the pathogenic examination of lung tissue. Qingyitang Dexamethasone, Santonstadine could decrease the levels of serum PBEF, the expression of PBEF mRNA and its protein in lung tissue, eliminate the extent of lung injury, but Linglanxin could not do that well.
The decline of endotoxin, TNF-α, IL-1β, IL-8 in Qingyitang group was more significant(p<0.05). Santonstadine could reduce amylase content obviously. And Dexamethasone showed superiority in ameliorating lung W/D ratio、PaCO2 and PaO2.
Conclusions:
Infusion of 1.5% sodium deoxycholate into biliopancreatic duct of Spraghe-Dawley rats, the pathologic changes of lung were edema and congestive with large of neutrial cells infiltration and focal atelectasis, the levels of serum AMY, PaCO2 and W/D increased obviously, meanwhile PaO2 decreased significantly. All indexes showed that the APALI model is welldone.
During acute lung injury induced by severe acute pancreatitis, many factors have very important effects on pathogenisis of ALI, such as elevated level of endotoxin,overexpression of PBEF which activated overexpression of TNF-a, IL-1β, IL-8.
Qingyitang could exert protecting effects through decreasing PBEF expression,reducing the excessive activation of alveolar macrophages, expression of TNF-a, IL-1βand IL-8 in a large number of inflammatory factors, aggregation of PMN, the level of serum endotoxin and amylase and decreasing alveolar permeability and other aspects of mechanism.
Dexamethasone and Santonstadine could play their available role in ALI by eliminating PBEF of different levels. Both of them could impact different indicators through different ways.
The effects of Chinese and Western medicines on SAP were related to different aspects of the pathogenesis of ALI, and play an important role by their own strengths. Combining Chinese and Western medicine under the guidance of the integration of traditional and western medicine theory, will provide a new effective way to prevent and cure SAP and SAP associated ALI in clinic.
引文
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33. Adams JM, Hauser CJ, Livingston DH, Lavery RF, Fekete Z, Deitch EA. Early trauma polymorphonuclear neutrophil responses to chemokines are associated with development of sepsis, pneumonia, and organ failure. J Trauma.2001, 51:452-456.
34. Hudson LD. Causes of the adult respiratory distress syndrome—clinical recognition. Clin Chest Med 1982,3:195-212.
35. Ware LB. Prognostic determinants of acute respiratory distress syndrome in adults:impact on clinical trial design. Crit Care Med.2005,33(Suppl):217-222.
36. Modelska K, Pittet JF, Folkesson HG, Courtney V, Matthay MA. Acid-induced lung injury. Protective effect of anti-interleukin-8 pretreatment on alveolar epithelial barrier function in rabbits. Am J Respir Crit Care Med.1999, 160:1450-1456.
37. Dudek SM, Garcia JG. Cytoskeletal regulation of pulmonary vascular permeability. J Appl Physiol,2001,91:1487-1500.
38. Ye SQ, Zhang LQ, Adyshev D, Usatyuk PV, Garcia AN, Lavoie TL, Verin AD, Natarajan V, Garcia JG. Pre-B-cell colony-enhancing factor is critically involved in thrombin-induced lung endothelial cell barrier dysregulation. Microvasc. Res. 2005,70,142-151.
39. Interactions between PBEF and oxidative stress proteins-A potential new mechanism underlying PBEF in the pathogenesis of acute lung injury L.Q. Zhang et al./FEBS Letters 582,2008,1802-1808.
40. Krzyznowska K, Krugluger W et al:Increased visfatin contrations in women with gestational diabetes mellitus. Clin Sci (Lond),2006,110:605-609.
41. Dahl, TB, Yndestad A, Skjellnad M, et al.Incersaed expression of Visfatin in macrophages of human unstable carotid and cornaryatherosclerosis:possible role in inflammation and plaque destabilization[J].Circulation,2007,115(8):972.
42.王培Visfatin/Nampt作为新的血管活性物质以及作为脑卒中防治新靶标的研究.第二军医大学2009年博士学位论文。
43. Brentano F; Schorr O; Ospelt C;Stanczyk J; Gay RE;Gay S;Kyburz D Pre-B cell colony-enhancing factor/visfatin, a new marker of inflammation in rheumatoid arthritis with proinflammatory and matrix-degrading activities. Arthritis Rheum 2007,56(9):2829-2839.
44.蓝丹前B细胞集落促进因子诱导正常人及重型先天性中性粒细胞减少症患者做系细胞分化功能研究.广西医科大学2006年博士学位论文。
1.张志宏,徐肇敏.急性胰腺炎.于中麟主编,消化病学精要系列丛书胰腺疾病。2005年3月第一版,沈阳辽宁科学技术出版社2005：1 84-189。
2. Samal B, Sun Y, Stearns G, Xie C, Suggs S, Mc Niece I. Cloning and characterization of the cDNA encoding a novel human pre-B-cell colony-enhancing factor. Mol Cell Biol.1994,14:1431-1437.
3. Nemeth E, Tashima LS, Yu Z, Bryant-Greenwood GD. Fetal membrane distention: I. Differentially expressed genes regulated by acute distention in amniotic epithelial (WISH) cells. Am J Obstet Gynecol.2000,182,50-59.57.
4. Ognjanovic S, Bao S, Yamamoto SY, et al. Genomic organization of the gene coding for human pre-B-cell colony enhancing factor and expression in human fetal membranes [J]. J Mol Endocrinol,2001,26 (2):107-117.
5. Marvin K W, Keelan JA, Eykholt RL, Sato TA, Mitchell MD. Use of cDNA arrays to generate differential expression profiles for inflammatory genes in human gestational membranes delivered at term and preterm. Mol Hum Reprod.2002; 8: 399-408.
6. Jia SH, Li Y, Parodo J, et al. Pre-B cell colony-enhancing factor inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis. J Clin Invest 2004; 113:1318-1327.
7. Peng Liu, Hailong Li,et al.Critical role of PBEF expression in pulmonary cell inflammation and permeability [J] Cell Biology International (2009) 33,19-30.
8. Ye SQ, Simon BA, Maloney JP, et al. Pre-B-cell colony enhancing factor as a potential novel biomarker in acute lung injury. Am J Respir Crit Care Med, 2005,171:361-370.
9. Moschen AR, Kaser A, Enrich B, et al. Visfatin, an adipocykiine with proinflammatory and immunomodulating properties[J].Immunol,2007, 178(3): 1748.
10.孙卫民,王惠琴.细胞因子研究方法学.北京：人民卫生出版社,1999.,592-623。
11.肖贞良,钱贵生,孙耕耘.TNF-α对大鼠肺微血管内皮细胞p-受体及其相关GRKs的影响.第三军医大学学报,2004,26(10)：859-862。
12.蔡栩栩, 韩玉昆.肺血管内皮细胞与急性肺损伤.国外医学儿科学分册,2002,29(1)：34-36。
13.金惠铭,刘清行,曹翔等.TNF-α引起的微血管内皮细胞功能障碍及其细胞分
子机制.微循环学杂志,2000,10(3)：5-6。
14. Wang HM, Ding RG, Ruan JX. Progress of the study on effect mechanism of neutrophils in acute lung injury. Chin J Ind Med,2002,15(6):348-350.
15. Osman MO, Kristrensen JU, Jacosen NO, et al. A, momoclonal anti-interleukin 8antibody(WS-4) inhibits cytokine response and acute lung injury in experimental severe acute necrosising pancreatitis in rabbits. Gut,1998, 43:232-239.
16. Deitch EA.Multiple systemic organ failure. Arch Surg 1992; 216:118-136.
17. Windsor JA;fearon K.C; Ross JA;Barclay GR; Smyth E; Poxton I; Garden OJ; Carter DC. Rale of serum endotoxin and antiendotoxin core antibody levels in predicting the development of multile organ failure in acute pancreatitis.Br J Surg 1993 Aug,80(8):1042-6.
18.姚咏明,施志国,田惠民.内毒素在多系统器官功能衰竭中的作用。国外医学创伤与外科基本问题分册。1992；6(1)：69-72。
19.陈海龙,周俊元.中西医结合治疗内毒素血症。中西医结合杂志,1991；11(3)：184-187。
20. Jacob AI; Goldberg pk; Bloom N; Degeshein GA; Kozinn PJ. Endotoxin and bacteria in portal blood.Gastroenterology 1997 Jun,72(6):1268-1270.
21. Navaratnam RL;Morris SE; Traber DL; Flynn J;Woodsan L;Linares H; Herndon DN.Endotoxin(LPS) increases mesenteric vascular resistance (MVR) and bacterial translocation (BT). J Trauma,1990,30(9):1104-1113.
22.李春盛,周景,杜培春,何新华.大黄对内毒素诱导致急性肺损伤的保护作用.中国中西医结合急救杂志,2000,7(1)：13-16。
23.王祥瑞,杭燕南.主编.急性肺损伤—基础与临床(第一版)。北京,中国协和医科大学出版社,2005：25。
24.中国中西医结合普通外科专业委员会重症急性胰腺炎中西医结合诊治常规(草案)。中国中西医结合外科杂志,2007,13(3)：232-237。
25.吴咸中.腹部外科实践(第三版)[M].天津：天津科学技术出版社,2004：1242-1262。
26.高振明,陈海龙,王立明,等。清胰汤对大鼠急性胰腺炎肺损伤中水通道蛋白-1表达的影响[J]。中国中西医结合外科杂志,2007,13(5)：460-463。
27.李海龙,陈海龙。核因子—KB在急性胰腺炎肺损伤发病机理中的作用及清胰汤影响的实验研究[D]大连医科大学2003年博士论文。
28. Andreasson S, Smith L, Risberg B. The role of high dose corticosteroids (HDC) in pulmonary capillary-alveolar membrane integrity. Acta Chir Scand Suppl 1985,526:83-93.
29.闻庆平,陈海龙,关凤林.清胰汤对大鼠重症急性胰腺炎时急性肺损伤治疗作用的观察[J].中国中西医结合外科杂志,2003,8(9)：302-306。
30. Sharma VK, Howden CW. Prophylactic antibiotic administration reduces sepsis and moaality in acute necrotizing pancreatitis:a meta-analysis [J]. Pancreas, 2001,22(1):28—31.
31. Heinrich S, Sehafer M, Rousson V, et al. Evidence—based treatment of acute pancreatitis:a look at established paradigms[J]. Ann Surg,2006,243(2): 154—168.
32.脱红芳,胁见裕.日本急性胰腺炎发病及诊疗现状[J].胰腺病学,2004,4(4)：254。
33.吴咸中.急性胰腺炎的中西医结合治疗。世界华人消化杂志200 1 9(4)：417-418。
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33. Adams JM, Hauser CJ, Livingston DH, Lavery RF, Fekete Z, Deitch EA. Early trauma polymorphonuclear neutrophil responses to chemokines are associated with development of sepsis, pneumonia, and organ failure. J Trauma.2001, 51:452-456.
34. Hudson LD. Causes of the adult respiratory distress syndrome—clinical recognition. Clin Chest Med 1982,3:195-212.
35. Ware LB. Prognostic determinants of acute respiratory distress syndrome in adults:impact on clinical trial design. Crit Care Med.2005,33(Suppl):217-222.
36. Modelska K, Pittet JF, Folkesson HG, Courtney V, Matthay MA. Acid-induced lung injury. Protective effect of anti-interleukin-8 pretreatment on alveolar epithelial barrier function in rabbits. Am J Respir Crit Care Med.1999, 160:1450-1456.
37. Dudek SM, Garcia JG. Cytoskeletal regulation of pulmonary vascular permeability. J Appl Physiol,2001,91:1487-1500.
38. Ye SQ, Zhang LQ, Adyshev D, Usatyuk PV, Garcia AN, Lavoie TL, Verin AD, Natarajan V, Garcia JG. Pre-B-cell colony-enhancing factor is critically involved in thrombin-induced lung endothelial cell barrier dysregulation. Microvasc. Res. 2005,70,142-151.
39. Interactions between PBEF and oxidative stress proteins-A potential new mechanism underlying PBEF in the pathogenesis of acute lung injury L.Q. Zhang et al./FEBS Letters 582,2008,1802-1808.
40. Krzyznowska K, Krugluger W et al:Increased visfatin contrations in women with gestational diabetes mellitus. Clin Sci (Lond),2006,110:605-609.
41. Dahl, TB, Yndestad A, Skjellnad M, et al.Incersaed expression of Visfatin in macrophages of human unstable carotid and cornaryatherosclerosis:possible role in inflammation and plaque destabilization[J].Circulation,2007,115(8):972.
42.王培Visfatin/Nampt作为新的血管活性物质以及作为脑卒中防治新靶标的研究.第二军医大学2009年博士学位论文。
43. Brentano F; Schorr O; Ospelt C;Stanczyk J; Gay RE;Gay S;Kyburz D Pre-B cell colony-enhancing factor/visfatin, a new marker of inflammation in rheumatoid arthritis with proinflammatory and matrix-degrading activities. Arthritis Rheum 2007,56(9):2829-2839.
44.蓝丹前B细胞集落促进因子诱导正常人及重型先天性中性粒细胞减少症患者做系细胞分化功能研究.广西医科大学2006年博士学位论文。
1.张志宏,徐肇敏.急性胰腺炎.于中麟主编,消化病学精要系列丛书胰腺疾病。2005年3月第一版,沈阳辽宁科学技术出版社2005：1 84-189。
2. Samal B, Sun Y, Stearns G, Xie C, Suggs S, Mc Niece I. Cloning and characterization of the cDNA encoding a novel human pre-B-cell colony-enhancing factor. Mol Cell Biol.1994,14:1431-1437.
3. Nemeth E, Tashima LS, Yu Z, Bryant-Greenwood GD. Fetal membrane distention: I. Differentially expressed genes regulated by acute distention in amniotic epithelial (WISH) cells. Am J Obstet Gynecol.2000,182,50-59.57.
4. Ognjanovic S, Bao S, Yamamoto SY, et al. Genomic organization of the gene coding for human pre-B-cell colony enhancing factor and expression in human fetal membranes [J]. J Mol Endocrinol,2001,26 (2):107-117.
5. Marvin K W, Keelan JA, Eykholt RL, Sato TA, Mitchell MD. Use of cDNA arrays to generate differential expression profiles for inflammatory genes in human gestational membranes delivered at term and preterm. Mol Hum Reprod.2002; 8: 399-408.
6. Jia SH, Li Y, Parodo J, et al. Pre-B cell colony-enhancing factor inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis. J Clin Invest 2004; 113:1318-1327.
7. Peng Liu, Hailong Li,et al.Critical role of PBEF expression in pulmonary cell inflammation and permeability [J] Cell Biology International (2009) 33,19-30.
8. Ye SQ, Simon BA, Maloney JP, et al. Pre-B-cell colony enhancing factor as a potential novel biomarker in acute lung injury. Am J Respir Crit Care Med, 2005,171:361-370.
9. Moschen AR, Kaser A, Enrich B, et al. Visfatin, an adipocykiine with proinflammatory and immunomodulating properties[J].Immunol,2007, 178(3): 1748.
10.孙卫民,王惠琴.细胞因子研究方法学.北京：人民卫生出版社,1999.,592-623。
11.肖贞良,钱贵生,孙耕耘.TNF-α对大鼠肺微血管内皮细胞p-受体及其相关GRKs的影响.第三军医大学学报,2004,26(10)：859-862。
12.蔡栩栩, 韩玉昆.肺血管内皮细胞与急性肺损伤.国外医学儿科学分册,2002,29(1)：34-36。
13.金惠铭,刘清行,曹翔等.TNF-α引起的微血管内皮细胞功能障碍及其细胞分
子机制.微循环学杂志,2000,10(3)：5-6。
14. Wang HM, Ding RG, Ruan JX. Progress of the study on effect mechanism of neutrophils in acute lung injury. Chin J Ind Med,2002,15(6):348-350.
15. Osman MO, Kristrensen JU, Jacosen NO, et al. A, momoclonal anti-interleukin 8antibody(WS-4) inhibits cytokine response and acute lung injury in experimental severe acute necrosising pancreatitis in rabbits. Gut,1998, 43:232-239.
16. Deitch EA.Multiple systemic organ failure. Arch Surg 1992; 216:118-136.
17. Windsor JA;fearon K.C; Ross JA;Barclay GR; Smyth E; Poxton I; Garden OJ; Carter DC. Rale of serum endotoxin and antiendotoxin core antibody levels in predicting the development of multile organ failure in acute pancreatitis.Br J Surg 1993 Aug,80(8):1042-6.
18.姚咏明,施志国,田惠民.内毒素在多系统器官功能衰竭中的作用。国外医学创伤与外科基本问题分册。1992；6(1)：69-72。
19.陈海龙,周俊元.中西医结合治疗内毒素血症。中西医结合杂志,1991；11(3)：184-187。
20. Jacob AI; Goldberg pk; Bloom N; Degeshein GA; Kozinn PJ. Endotoxin and bacteria in portal blood.Gastroenterology 1997 Jun,72(6):1268-1270.
21. Navaratnam RL;Morris SE; Traber DL; Flynn J;Woodsan L;Linares H; Herndon DN.Endotoxin(LPS) increases mesenteric vascular resistance (MVR) and bacterial translocation (BT). J Trauma,1990,30(9):1104-1113.
22.李春盛,周景,杜培春,何新华.大黄对内毒素诱导致急性肺损伤的保护作用.中国中西医结合急救杂志,2000,7(1)：13-16。
23.王祥瑞,杭燕南.主编.急性肺损伤—基础与临床(第一版)。北京,中国协和医科大学出版社,2005：25。
24.中国中西医结合普通外科专业委员会重症急性胰腺炎中西医结合诊治常规(草案)。中国中西医结合外科杂志,2007,13(3)：232-237。
25.吴咸中.腹部外科实践(第三版)[M].天津：天津科学技术出版社,2004：1242-1262。
26.高振明,陈海龙,王立明,等。清胰汤对大鼠急性胰腺炎肺损伤中水通道蛋白-1表达的影响[J]。中国中西医结合外科杂志,2007,13(5)：460-463。
27.李海龙,陈海龙。核因子—KB在急性胰腺炎肺损伤发病机理中的作用及清胰汤影响的实验研究[D]大连医科大学2003年博士论文。
28. Andreasson S, Smith L, Risberg B. The role of high dose corticosteroids (HDC) in pulmonary capillary-alveolar membrane integrity. Acta Chir Scand Suppl 1985,526:83-93.
29.闻庆平,陈海龙,关凤林.清胰汤对大鼠重症急性胰腺炎时急性肺损伤治疗作用的观察[J].中国中西医结合外科杂志,2003,8(9)：302-306。
30. Sharma VK, Howden CW. Prophylactic antibiotic administration reduces sepsis and moaality in acute necrotizing pancreatitis:a meta-analysis [J]. Pancreas, 2001,22(1):28—31.
31. Heinrich S, Sehafer M, Rousson V, et al. Evidence—based treatment of acute pancreatitis:a look at established paradigms[J]. Ann Surg,2006,243(2): 154—168.
32.脱红芳,胁见裕.日本急性胰腺炎发病及诊疗现状[J].胰腺病学,2004,4(4)：254。
33.吴咸中.急性胰腺炎的中西医结合治疗。世界华人消化杂志200 1 9(4)：417-418。