血小板活化因子受体拮抗剂BN52021在大鼠重症急性胰腺炎早期病程中的作用和机制
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摘要
目的
探讨血小板活化因子受体(PAF-R)拮抗剂BN52021在大鼠重症急性胰腺炎(SAP)早期病程中的作用和机制。
方法
1.用免疫组织化学染色方法研究PAF-R在Wistar大鼠胰腺的定位和分布。
2.用胰胆管插入逆行注射牛磺胆酸钠制备SAP大鼠模型;用药代动力学方法研究BN52021治疗大鼠SAP的最佳量效关系。
3.用RT-PCR和Western blot方法分别研究PAF-R和核因子κB单体(NF-κBp65) mRNA和蛋白质在SAP大鼠胰腺组织的动态变化以及BN52021的影响。
结果
1.冰冻切片中显示在Wistar大鼠正常胰腺组织微血管内皮细胞有PAF-R表达;石蜡切片中显示胰岛细胞有PAF-R表达,主要分布于细胞浆和细胞核,核膜着色较深,但是血管内皮细胞着色较浅。
2.血清淀粉酶、PLA2、腹水、胰腺组织病理学和胰腺组织PAF-R mRNA表达的结果显示在2.5 mg/kg、5.0 mg/kg、10.0 mg/kg BN52021三个剂量中,5.0mg/kg BN52021静脉注射给Wistar大鼠治疗SAP具有最佳的量效关系(P<0.05);这些结果也显示SAP模型制备是成功的。
3.PAF-R mRNA结果显示,SAP模型(SAP组)和BN52021治疗后(BN组)均是动态变化的,早期逐渐上升,在3h达到高峰,以后逐渐降低,除1h和2h外其余各时相点与假手术动物(SO组)相比有显著性差异(P<0.05),除6h外BN组与SAP组各时相点之间无显著性差异;PAF-R蛋白质结果显示,SAP组和BN组PAF-R蛋白质与PAF-R mRNA的变化趋势基本是一致的,先升后降,除1h外其余各时相点与SO组有显著性差异(P<0.05),BN组与SAP组各时相点之间无显著性差异。
4.NF-κBp65mRNA结果显示,SAP组和BN组均是动态变化的,呈双峰改变,SAP组双峰顶分别在1h和24h,BN组双峰顶分别在1h和12h,于6h降至最低,SAP组在2h、3h、12h和24h较SO组显著增加(P<0.05),BN组在1h较SAP组显著增加(P<0.05),BN组各时相点较SO组显著升高(P<0.05);NF-?Bp65蛋白质结果显示,SAP组在1h,3h和6h较SO组显著升高(P<0.01),在2h,12h和24h较SO组也显著升高(P<0.05),BN组各时相点较SO组显著升高(P<0.05),在1h,3h和6h较SAP组显著降低(P<0.05)。
结论
1.研究发现PAF-R不仅定位分布于Wistar大鼠胰腺微血管内皮细胞,也定位分布于其胰腺胰岛细胞胞浆和细胞核内。
2.经胰胆管插入逆行注射牛磺胆酸钠制备SAP大鼠模型是可靠的;用5.0mg/kg BN52021静脉注射治疗Wistar大鼠SAP具有最佳的量效关系。
3.PAF-R在SAP大鼠胰腺组织表达是动态变化的,PAF-R可能参与了SAP的发病机制,对SAP病情的发生发展起一定的作用;BN52021可降低SAP血清淀粉酶和改善其病理变化,但对胰腺组织PAF-R的表达无显著影响。分析认为,BN52021治疗SAP大鼠并不是通过调控胰腺组织PAF-R的表达量,而是通过竞争性抑制上升的PAF与同时表达量上升的PAF-R结合,从而影响PAF-R的信号转导,实现它的治疗作用。
4.NF-kB是PAF-R信号转导通路下游的关键成分。NF-kBp65在SAP大鼠胰腺组织表达是动态变化的,NF-kBp65可能参与了SAP的发病机制,对SAP病情的发生发展起一定的作用;BN52021在早期可降低NF-κBp65蛋白质的表达,从而发挥一定的治疗作用。
AIM
This research is to investigate the role and mechanism of platelet activating factor receptor (PAF-R) antagonist (BN52021, Ginkgolide B) in early stage of severe acute pancreatitis (SAP) in rats.
METHOD
1.The method of immunity hisochemistry staining was employed to study the location and distribution of PAF-R in normal pancreas tissues of Wistar rats.
2.The sodium aurocholatet was injected through pancreaticobiliary duct in retrograde direction to establish SAP model and the method of pharmacokinetics was used to explore the optimal dosage of BN52021 on rats with SAP.
3.The methods of PT-PCR and Western blot were adopted respectively to investigate the changes of PAF-R and NF-κBp65 mRNA and proteins expression in pancreatic tissues of rats and the effect of BN52021.
RESULTS
1.In frozen section PAF-R located in pancreatic microvascular endothelial cells. In paraffin section PAF-R located in Pancreas islet cytoplasm and cell nucleus; there was little colouration in pancreatic microvascular endothelial cells.
2.The results of the serum amylase, PLA2, ascites, pathologic score and PAF-RmRNA expression showed that, compared with 2.5 mg.kg-1 and 10.0 mg.kg-1, 5.0 mg.kg-1 of BN52021 by vein injection was the optimal dosage in treating rats with SAP(P<0.05). The results also showed that SAP model was successful.
3.PAF-mRNA showed dynamic changes in SAP and BN groups. It increased gradually in early stage, reached a peak at 3h, and decreased gradually later. There were significant differences between PAF-mRNA in SAP and BN groups and that in SO groups at each time point except at 1h and 2h (P<0.05), whereas there was no significant difference between PAF-mRNA in BN groups and that in SAP groups at each time point except at 6h. The changes of PAF-R protein in SAP and BN groups shared the similar pattern with those of PAF-R mRNA. There were significant differences between PAF-R protein in SAP and BN groups and that in SO groups at each time point except at 1h(P<0.05), whereas there was no significant difference between PAF-R protein in BN groups and that in SAP groups.
4.The expression of NF-κBp65 mRNA dynamically changed in both SAP and BN groups. The mRNA level was higher in SAP groups than that in SO groups at 2h, 3h, 12h, and 24h after operation (P<0.05). The mRNA level was higher in BN groups than that in SO groups at each time point (P<0.05), and also higher than that in SAP groups at 1 h (P<0.05). The NF-κBp65 protein level was higher in SAP groups than that in SO groups at 1h, 3h, and 6h (P<0.01), and 2h, 12h, and 24h (P<0.05). The NF-κBp65 protein level was higher in BN groups than that in SO groups at all time points (P<0.05), and lower in BN groups than that in SAP groups at 1h, 3h, and 6h (P<0.05).
CONCLUSIONS
1. It is found that PAF-R is distributed not only in pancreatic microvascular endothelial cells but also in pancreatic islet cells of Wistar rats.
2. It is found that 5.0 mg.kg-1 of BN52021 by vein injection is the optimal dosage in treating rats with SAP.
3. PAF-R is found to play an important role in occurrence and development of SAP. BN52021 exerts biological effects through the combination the increase of PAF expression achieved through competitive inhibition and the simultaneous increase of PAF-R expression rather than through regulating PAF-R expression in pancreatic tissues.
4. The NF-κB is a key factor in the lower reaches of signal transduction pathways of PAF-R. The expression of NF-κBp65 in pancreatic tissues dynamically changed and the changing might have played a role in pathogenesis of SAP. BN52021 exerted therapeutic effect by reducing the expression level of NF-κBp65 protein in the early stage of SAP.
探讨血小板活化因子受体(PAF-R)拮抗剂BN52021在大鼠重症急性胰腺炎(SAP)早期病程中的作用和机制。
方法
1.用免疫组织化学染色方法研究PAF-R在Wistar大鼠胰腺的定位和分布。
2.用胰胆管插入逆行注射牛磺胆酸钠制备SAP大鼠模型;用药代动力学方法研究BN52021治疗大鼠SAP的最佳量效关系。
3.用RT-PCR和Western blot方法分别研究PAF-R和核因子κB单体(NF-κBp65) mRNA和蛋白质在SAP大鼠胰腺组织的动态变化以及BN52021的影响。
结果
1.冰冻切片中显示在Wistar大鼠正常胰腺组织微血管内皮细胞有PAF-R表达;石蜡切片中显示胰岛细胞有PAF-R表达,主要分布于细胞浆和细胞核,核膜着色较深,但是血管内皮细胞着色较浅。
2.血清淀粉酶、PLA2、腹水、胰腺组织病理学和胰腺组织PAF-R mRNA表达的结果显示在2.5 mg/kg、5.0 mg/kg、10.0 mg/kg BN52021三个剂量中,5.0mg/kg BN52021静脉注射给Wistar大鼠治疗SAP具有最佳的量效关系(P<0.05);这些结果也显示SAP模型制备是成功的。
3.PAF-R mRNA结果显示,SAP模型(SAP组)和BN52021治疗后(BN组)均是动态变化的,早期逐渐上升,在3h达到高峰,以后逐渐降低,除1h和2h外其余各时相点与假手术动物(SO组)相比有显著性差异(P<0.05),除6h外BN组与SAP组各时相点之间无显著性差异;PAF-R蛋白质结果显示,SAP组和BN组PAF-R蛋白质与PAF-R mRNA的变化趋势基本是一致的,先升后降,除1h外其余各时相点与SO组有显著性差异(P<0.05),BN组与SAP组各时相点之间无显著性差异。
4.NF-κBp65mRNA结果显示,SAP组和BN组均是动态变化的,呈双峰改变,SAP组双峰顶分别在1h和24h,BN组双峰顶分别在1h和12h,于6h降至最低,SAP组在2h、3h、12h和24h较SO组显著增加(P<0.05),BN组在1h较SAP组显著增加(P<0.05),BN组各时相点较SO组显著升高(P<0.05);NF-?Bp65蛋白质结果显示,SAP组在1h,3h和6h较SO组显著升高(P<0.01),在2h,12h和24h较SO组也显著升高(P<0.05),BN组各时相点较SO组显著升高(P<0.05),在1h,3h和6h较SAP组显著降低(P<0.05)。
结论
1.研究发现PAF-R不仅定位分布于Wistar大鼠胰腺微血管内皮细胞,也定位分布于其胰腺胰岛细胞胞浆和细胞核内。
2.经胰胆管插入逆行注射牛磺胆酸钠制备SAP大鼠模型是可靠的;用5.0mg/kg BN52021静脉注射治疗Wistar大鼠SAP具有最佳的量效关系。
3.PAF-R在SAP大鼠胰腺组织表达是动态变化的,PAF-R可能参与了SAP的发病机制,对SAP病情的发生发展起一定的作用;BN52021可降低SAP血清淀粉酶和改善其病理变化,但对胰腺组织PAF-R的表达无显著影响。分析认为,BN52021治疗SAP大鼠并不是通过调控胰腺组织PAF-R的表达量,而是通过竞争性抑制上升的PAF与同时表达量上升的PAF-R结合,从而影响PAF-R的信号转导,实现它的治疗作用。
4.NF-kB是PAF-R信号转导通路下游的关键成分。NF-kBp65在SAP大鼠胰腺组织表达是动态变化的,NF-kBp65可能参与了SAP的发病机制,对SAP病情的发生发展起一定的作用;BN52021在早期可降低NF-κBp65蛋白质的表达,从而发挥一定的治疗作用。
AIM
This research is to investigate the role and mechanism of platelet activating factor receptor (PAF-R) antagonist (BN52021, Ginkgolide B) in early stage of severe acute pancreatitis (SAP) in rats.
METHOD
1.The method of immunity hisochemistry staining was employed to study the location and distribution of PAF-R in normal pancreas tissues of Wistar rats.
2.The sodium aurocholatet was injected through pancreaticobiliary duct in retrograde direction to establish SAP model and the method of pharmacokinetics was used to explore the optimal dosage of BN52021 on rats with SAP.
3.The methods of PT-PCR and Western blot were adopted respectively to investigate the changes of PAF-R and NF-κBp65 mRNA and proteins expression in pancreatic tissues of rats and the effect of BN52021.
RESULTS
1.In frozen section PAF-R located in pancreatic microvascular endothelial cells. In paraffin section PAF-R located in Pancreas islet cytoplasm and cell nucleus; there was little colouration in pancreatic microvascular endothelial cells.
2.The results of the serum amylase, PLA2, ascites, pathologic score and PAF-RmRNA expression showed that, compared with 2.5 mg.kg-1 and 10.0 mg.kg-1, 5.0 mg.kg-1 of BN52021 by vein injection was the optimal dosage in treating rats with SAP(P<0.05). The results also showed that SAP model was successful.
3.PAF-mRNA showed dynamic changes in SAP and BN groups. It increased gradually in early stage, reached a peak at 3h, and decreased gradually later. There were significant differences between PAF-mRNA in SAP and BN groups and that in SO groups at each time point except at 1h and 2h (P<0.05), whereas there was no significant difference between PAF-mRNA in BN groups and that in SAP groups at each time point except at 6h. The changes of PAF-R protein in SAP and BN groups shared the similar pattern with those of PAF-R mRNA. There were significant differences between PAF-R protein in SAP and BN groups and that in SO groups at each time point except at 1h(P<0.05), whereas there was no significant difference between PAF-R protein in BN groups and that in SAP groups.
4.The expression of NF-κBp65 mRNA dynamically changed in both SAP and BN groups. The mRNA level was higher in SAP groups than that in SO groups at 2h, 3h, 12h, and 24h after operation (P<0.05). The mRNA level was higher in BN groups than that in SO groups at each time point (P<0.05), and also higher than that in SAP groups at 1 h (P<0.05). The NF-κBp65 protein level was higher in SAP groups than that in SO groups at 1h, 3h, and 6h (P<0.01), and 2h, 12h, and 24h (P<0.05). The NF-κBp65 protein level was higher in BN groups than that in SO groups at all time points (P<0.05), and lower in BN groups than that in SAP groups at 1h, 3h, and 6h (P<0.05).
CONCLUSIONS
1. It is found that PAF-R is distributed not only in pancreatic microvascular endothelial cells but also in pancreatic islet cells of Wistar rats.
2. It is found that 5.0 mg.kg-1 of BN52021 by vein injection is the optimal dosage in treating rats with SAP.
3. PAF-R is found to play an important role in occurrence and development of SAP. BN52021 exerts biological effects through the combination the increase of PAF expression achieved through competitive inhibition and the simultaneous increase of PAF-R expression rather than through regulating PAF-R expression in pancreatic tissues.
4. The NF-κB is a key factor in the lower reaches of signal transduction pathways of PAF-R. The expression of NF-κBp65 in pancreatic tissues dynamically changed and the changing might have played a role in pathogenesis of SAP. BN52021 exerted therapeutic effect by reducing the expression level of NF-κBp65 protein in the early stage of SAP.
引文
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46 Korth R, Benveniste J. BN 52021 displaces [3H]paf-acether from, and inhibits its binding to intact human platelets. Eur J Pharmacol 1987; 142: 331-341
47 Marrache AM, Gobeil F JR[0], Bernier SG, , et al. Proinflammatory gene induction by platelet-activating factor mediated via its cognate nuclear receptor. J Immunol 2002; 169: 6474-6481
48夏时海,全金梅,乔爱军,等.生长抑素衍生物善宁加倍剂量治疗重症急性胰腺炎的疗效研究.世界华人消化杂志, 2002,10(10):1157-1161
49 Baker RR, Chang HY. Alkylglycerophosphate acetyltransferase and lyso platelet activating factor acetyltransferase, two key enzymes in the synthesisof platelet activating factor, are found in neuronal nuclei isolated from cerebral cortex. Biochim Biophys Acta 1996; 1302: 257-263
50 Schievella AR, Regier MK, Smith WL, et al. Calciummediated translocation of cytosolicphospholipase A2 to the nuclear envelope and endoplasmic reticulum. J Biol Chem 1995; 270: 30749-30754
51 Dagenais P, Thivierge M, Stankova J, , et al. Modulation of platelet-activating factor receptor (PAFR) gene expression via NFkB in MonoMac-1 cells. Inflamm Res 1997; 46: s161-162
52 Nakao A, Watanabe T , Bitoh H. cAMP mediates homologous downregulation of PAF receptor mRNA expression in mesangial cells Am J Physiol 1997;273(3 pt 2):445-450
53 Thivierge M, Parent JL, Stankova J, et al. Modulation of human platelet-activating factor receptor gene expression protein kinase C activation. J Immunol 1996; 157: 4681-4687
54 Hourton D, Stengel D, Chapman MJ, et al. Oxidized low density lipoproteins down regulate LPS-induced platelet-activating factor receptor expression in human monocyte-derived macrophages: implications for LPS-induced nuclear factor-kappaB binding activity. Eur J Biochem 2001; 268: 4489-4496
55 Gray KD, Simovic MO, Blackwell TS, et al. Activation of nuclear factor kappa B and severe hepatic necrosis may mediate systemic inflammation in choline- deficient/ethionine-supplemented diet-induced pancreatitis.Pancreas 2006; 33:260-267
56 Hosford D, Braquet P. Antagonists of platelet-activating factor: chemistry, pharmacology and clinical applications. Prog Med Chem 1990; 27: 325-380
57 Burgos RA, Hidalgo MA, Matthei SM, et al. Determination of specific receptor sites for platelet activating factor in bovine neutrophils. AmJ Vet Res 2004;65:628-636
58 Honda Z, Takano T, Gotoh Y, et al. Transfected Platelet activating factor receptor activates mitogen-activated protein (MAP) kinase and MAP kinase kinase in chinese hamster ovary cells. J Biol Chem 1994; 3: 2307-2315
59 Siebenlist U, Franzoso G, Brown K. Structure, regulation and function of NF-kappaB. Annu Rev Cell Biol, 1994, 10:405–455
60 Chaqour B, Howard PS, Richards CF, et al. Mechanical stretch induces platelet- activating-factor receptor gene expression through the NF-kappa B transcription factor. J Mol Cell Cardiol,1999,31: 1345–1355
61 Tak PP, Firestein GS. NF- B: a key role in inflammatory diseases. J Clin Invest, 2001,107:7–11
62 Butcher HL, Kennette WA, Collins O, et al. Metallothionein Mediates the Level andActivity of Nuclear Factor-B in Murine Fibroblasts . J Pharmacol Exp Ther, 2004,310: 589-598
63毕慧英,夏时海.核因子kB在急性胰腺炎全身反应综合症中的作用.世界华人消化杂志,2004,12(12):2837-2841
64夏时海,赵晓晏,郭萍.急性胰腺炎促炎细胞因子和抗炎细胞因子的动态变化及其作用.临床消化病杂志, 2001, 13:51-53
65 Dunn J A,Li C,Ha T,et al. Therapeutic modification of nuclear factor ?appaB binding activity and tumor necrosis factor-alpha gene expression during acute biliary pancreatitis. Am Sury,1997,63(12):1036-1043.
66 Blinman TA,Gukovsky I,Gukovskaya A,et al. Intra- pancreatic expression cytokines IL-6 and KC is increased in cerulein pancreatitis:regulation by NF-?appaB .Gastroenter- ology,1998,114(4P2):A443-449
67 Madhav B,Mark B,Sheila SK, et al. Inflammatory mediators in acute pancreatitis.Journal of pathology,2000;190(7):117-125
68 Chen Fei, Castranova V, Shi Xianglin, et al. New insights into the role of nuclear factor-?B,a Ubiquitous transcription factor in the initiation of diseases. Clinical Chemistry, 1999, 45(1):7-17
69 Nishimune H,Vasseur S,Wiese S,et al.Reg2 is a motoneuron neurotrophic factor and a singalling intermediate in the CNTF survival pathway.Nat Cell Biol,2000,2(12):906-914
70 Hietarranta A,Mustonnen H,Puolakkainen P,et al. Pro- inflammatory effects of pancreatic elastase are mediated through TLR4 and NF-?appaB.Biochem Biophys Res Commun, 2004, 323(1):192-196
71刘洪斌,崔乃强,李东华,等.磷脂酶A2与NF-?B在大鼠急性坏死性胰腺炎肺损伤中的作用.中国中西医结合外科杂志,2005,11(2):115-119
2 Flickinger BD, Olson MS. Localization of the platelet-activating factor receptor to rat pancreatic microvascular endothelial cells. Am J Pathol, 1999, 154(5):1353-1358
3 Liu LR, Xia SH. Role of platelet-activating factor in patho- genesis of acute pancreatitis.World J Gastroenterol,2006;12(4): 539-545
4夏国栋,夏时海.血小板活化因子受体的研究进展.世界华人消化杂志, 2005;13(3):381- 384
5 Marrache AM, Gobeil F JR, Bernier SG, et al. Proinflammatory gene induction by platelet-activating factor mediated via its cognate nuclear receptor. J Immunol, 2002, 169 (11): 6474-6481
6 Sugano T, Narahara H, Nasu K, et al. Effects of platelet-activating factor on cytokine production by human uterine cervical fibroblasts. Molecular Human Reproduction, 2001, 7(5): 475-481
7 Boccellino M, Biancone L, Cantaluppi V, et al. Effect of platelet-activating factor receptor expression on CHO cell motility. J Cell Physiol, 2000, 183(2): 254–264
8 Schievella AR, Regier MK, Smith WL, et al. Calciummediated translocation of cytosolic phospholipase A2 to the nuclear envelope and endoplasmic reticulum. J Biol Chem, 1995, 270(51): 30749-30754
9 Baker RR, Chang HY. Alkylglycerophosphate acetyltransferase and lyso platelet activating factor acetyltransferase, two key enzymes in the synthesisof platelet activating factor, are found in neuronal nuclei isolated from cerebral cortex. Biochim Biophys Acta, 1996, 1302(3): 257-263
10 Betu¨Lersoy, MD Afig HU¨Seyinovs? U¨Kran Darcan. The Role of Platelet-Activating Factor in Pathogenesis of Type 1 Diabetes. Diabetes Care, 2005, 28(4): 980
11 Beck JC, Goodner CJ, Wilson C, et al. Effects of ginkgolide B, a platelet-activating factor inhibitor on insulitis in the spontaneously diabetic BB rat. Autoimmunity, 1991, 9(3): 225-235
12夏时海,赵晓晏,郭萍.犬重症急性胰腺炎血液循环障碍及血小板活化因子受体拮抗剂的干预.世界华人消化杂志, 2001, 9(5): 550-554
13 Shirasaki H, Nishikawa M, Adcock IM, et al. Expression of platelet-activating factor receptor mRNA in human and guinea pig lung. Am J Respir Cell Mol Biol, 1994, 10(5): 533-537
14 Ibe BO, Portugal AM, Chaturvedi S, et al. Oxygen-dependent PAF receptor binding and intracellular signaling in ovine fetal pulmonary vascular smooth muscle. Am J Physiol Lung Cell Mol Physiol, 2005, 288(5): 879-886
15聂珍贵,王文杰.银杏内酯B对血小板活化因子刺激的大鼠中性粒细胞功能的影响.药学学报2003, 38(2): 98-102
16 Fang HY, Lin TS, Lin JP, et al. Cyclooxygenase-2 in human non-small cell lung cancer. Eur J Surg Oncol, 2003, 29(2): 171-177
17夏国栋,夏时海.血小板活化因子受体的研究进展.世界华人消化杂志, 2005,13(3): 381-384
18赵志伶,夏时海,陈虹.银杏苦内酯B的临床和药理学研究进展.武警医学, 2006, 17:132-135
19 Diserbo M, Cand F, Ziade M, et al. Stimulation of platelet-activating factor (PAF) receptors increases inositol phosphate production and cytosolic free Ca2+ concentrations in N1E-115 neuroblastoma cells. Cell Calcium, 1995, 17: 442-452
20 Collins LC, Roberts AM. Effects of platelet-activating factor on arteriolar and venular tone in rat trachea. Microvasc Res, 1997, 53: 63-72
21 Aho Hj, Koskensalo SM, Nevalainen TJ. Experimental pancreatitis in the rat.sodium taurocholate induced acute haemorrhagic pancreatitis. Scand J Gastroenterol,1980, 15: 411-416
22 Fourtillan JB, Brisson AM, Girault J. Pharmacokinetics of bilobalide, ginkgokide A and Ginkgolide B in healthy volunteers following oral and intracenous administrations of Ginkgo biloba extract(EGB761). Therapie, 2004, 50:137-144
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24 Wang G Y,Liu TJ,Yi WM,et al.Comparison among three types of experimental acute pancreatiyis animal model.ChinJ ExperSurg,1995,12(1):56-57
25 Yu YX,Gu ZY.Establishment of acute necrosis pancreatitis animal model by delivering sodium taurocholate intobilio pancreaticduct. Chin J Exper Surg,1996,13(5):261-262
26 Yao ZHX,Xu XY,Lin YY,et al.Establishment of severe acute pancreatitia animal modeland its mechanism research. hinJ ExperSurg,1997,12(4):229-231
27 Ding SP,Li JC,Jin C.Amouse model of severe acute pancreatitis induced with caerulein and lipopoly- saccharide.World J Gastroenterol. 2003, 9(3): 584-589
28王兴鹏,袁耀宗,徐家裕.银杏苦内酯对急性胰腺炎大鼠的治疗作用.中国药理学通报,1995,11:199-201
29冀振华,王本茂,李少华.血小板激活因子在急性重症胰腺炎发病过程中作用的实验研究.中华外科杂志, 1997,35:108-110
30冀振华,王本茂,李少华,等.血小板活化因子在急性胰腺炎大鼠肠系膜微循环障碍中的作用.中华医学杂志, 1995,75:139-140
31 Minor T, Isselhard W, Yamaguchi T. Involvement of platelet activating factor in microcirculatory disturbances after global hepatic ischemia. J Surg Res,1995, 58:536-540
32 Zhou W, McCollum MO, Levine BA, et al. Role of platelet-activating factor in pancreatitis-associated acute lung injury in the rat. Am J Pathol, 1992,140: 971-979
33 Bedirli A, Gokahmetoglu S, Sakrak O, et al. Beneficial effects of recombinant platelet-activating factor acetylhydrolase and BN52021 on bacterial translocation in cerulein-induced pancreatitis. Eur Surg Res, 2004,36:136-141
34邸瑶,夏时海,佟长青.银杏苦内酯B对重症急性胰腺炎大鼠血浆细胞因子的影响.世界华人消化杂志,2006,14:2169-2173
35 Redl H, Vogl C, Schiesser A, et al. Effect of the PAF antagonist BN52021 in ovine endotoxin shock. J Lipid Mediat, 1990, 2(S): 195-201
36 Lopez-Farre A, Gomez-Garre D, Beroabeu F, et al. Platelet-activating factor mediates glycerol-induced acute renal failure in rats. Clin Sci, 1990,79:551-558
37 Beck JC, Goodner CJ, Wilson C, et al. Effects of ginkgolide B, a platelet- activating factor inhibitor on insulitis in the spontaneously diabetic BB rat. Autoimmunity, 1991,9:225-235
38 Samuel I, Zaheer A, Fisher RA. In Vitro Evidence for Role of ERK, p38, and JNK in Exocrine Pancreatic Cytokine Production.J Gastrointest Surg 2006; 10:1376-83
39 Browne GW, Pitchumoni CS. Pathophysiology of pulmonary complications of acutepancreatitis.World J Gastroenterol 2006; 12:7087-7096
40 Sugano T, Narahara H, Nasu K, et al. Effects of platelet-activating factor on cytokine production by human uterine cervical fibroblasts. Molecular Human Reproduction 2001;7: 475-481
41 Boccellino M, Biancone L, Cantaluppi V, et al. Effect of platelet-activating factor receptor expression on CHO cell motility. J Cell Physiol 2000;183: 254–264
42 Seale JP, Nourshargh S, Hellewell PG, et al. Mechanism of action of platelet activating factor in the pulmonary circu-lation;an investigation using ovelisotopic system in rabbit isolated lung. Br J Parmacol 1991; 104: 251-257
43 Vincent JL, Spapen H, Bakker J, et al. PhaseⅡmulticenter clinical study of the platelet-activating factor receptor antagonist BB-882 in the treatment of sepsis. Crit Care Med 2000; 28:638-642
44 de Souza LJ, Sampietre SN, Assis RS, et al. Effect of platelet-activating factor antagonists (BN-52021, WEB-2170, and BB-882) on bacterial translocation in acute pancreatitis.J Gastrointest Surg 2001 ;5:364-370
45 Marques SA, Dy LC, Southall MD, et al. The platelet-activating factor receptor activates the extracellular signal-regulated kinase mitogen-activated protein kinase and induces proliferation of epidermal cells through an epidermal growth factor-receptor- dependent pathway. J Pharmacol Exp Ther 2002; 300:1026-1035
46 Korth R, Benveniste J. BN 52021 displaces [3H]paf-acether from, and inhibits its binding to intact human platelets. Eur J Pharmacol 1987; 142: 331-341
47 Marrache AM, Gobeil F JR[0], Bernier SG, , et al. Proinflammatory gene induction by platelet-activating factor mediated via its cognate nuclear receptor. J Immunol 2002; 169: 6474-6481
48夏时海,全金梅,乔爱军,等.生长抑素衍生物善宁加倍剂量治疗重症急性胰腺炎的疗效研究.世界华人消化杂志, 2002,10(10):1157-1161
49 Baker RR, Chang HY. Alkylglycerophosphate acetyltransferase and lyso platelet activating factor acetyltransferase, two key enzymes in the synthesisof platelet activating factor, are found in neuronal nuclei isolated from cerebral cortex. Biochim Biophys Acta 1996; 1302: 257-263
50 Schievella AR, Regier MK, Smith WL, et al. Calciummediated translocation of cytosolicphospholipase A2 to the nuclear envelope and endoplasmic reticulum. J Biol Chem 1995; 270: 30749-30754
51 Dagenais P, Thivierge M, Stankova J, , et al. Modulation of platelet-activating factor receptor (PAFR) gene expression via NFkB in MonoMac-1 cells. Inflamm Res 1997; 46: s161-162
52 Nakao A, Watanabe T , Bitoh H. cAMP mediates homologous downregulation of PAF receptor mRNA expression in mesangial cells Am J Physiol 1997;273(3 pt 2):445-450
53 Thivierge M, Parent JL, Stankova J, et al. Modulation of human platelet-activating factor receptor gene expression protein kinase C activation. J Immunol 1996; 157: 4681-4687
54 Hourton D, Stengel D, Chapman MJ, et al. Oxidized low density lipoproteins down regulate LPS-induced platelet-activating factor receptor expression in human monocyte-derived macrophages: implications for LPS-induced nuclear factor-kappaB binding activity. Eur J Biochem 2001; 268: 4489-4496
55 Gray KD, Simovic MO, Blackwell TS, et al. Activation of nuclear factor kappa B and severe hepatic necrosis may mediate systemic inflammation in choline- deficient/ethionine-supplemented diet-induced pancreatitis.Pancreas 2006; 33:260-267
56 Hosford D, Braquet P. Antagonists of platelet-activating factor: chemistry, pharmacology and clinical applications. Prog Med Chem 1990; 27: 325-380
57 Burgos RA, Hidalgo MA, Matthei SM, et al. Determination of specific receptor sites for platelet activating factor in bovine neutrophils. AmJ Vet Res 2004;65:628-636
58 Honda Z, Takano T, Gotoh Y, et al. Transfected Platelet activating factor receptor activates mitogen-activated protein (MAP) kinase and MAP kinase kinase in chinese hamster ovary cells. J Biol Chem 1994; 3: 2307-2315
59 Siebenlist U, Franzoso G, Brown K. Structure, regulation and function of NF-kappaB. Annu Rev Cell Biol, 1994, 10:405–455
60 Chaqour B, Howard PS, Richards CF, et al. Mechanical stretch induces platelet- activating-factor receptor gene expression through the NF-kappa B transcription factor. J Mol Cell Cardiol,1999,31: 1345–1355
61 Tak PP, Firestein GS. NF- B: a key role in inflammatory diseases. J Clin Invest, 2001,107:7–11
62 Butcher HL, Kennette WA, Collins O, et al. Metallothionein Mediates the Level andActivity of Nuclear Factor-B in Murine Fibroblasts . J Pharmacol Exp Ther, 2004,310: 589-598
63毕慧英,夏时海.核因子kB在急性胰腺炎全身反应综合症中的作用.世界华人消化杂志,2004,12(12):2837-2841
64夏时海,赵晓晏,郭萍.急性胰腺炎促炎细胞因子和抗炎细胞因子的动态变化及其作用.临床消化病杂志, 2001, 13:51-53
65 Dunn J A,Li C,Ha T,et al. Therapeutic modification of nuclear factor ?appaB binding activity and tumor necrosis factor-alpha gene expression during acute biliary pancreatitis. Am Sury,1997,63(12):1036-1043.
66 Blinman TA,Gukovsky I,Gukovskaya A,et al. Intra- pancreatic expression cytokines IL-6 and KC is increased in cerulein pancreatitis:regulation by NF-?appaB .Gastroenter- ology,1998,114(4P2):A443-449
67 Madhav B,Mark B,Sheila SK, et al. Inflammatory mediators in acute pancreatitis.Journal of pathology,2000;190(7):117-125
68 Chen Fei, Castranova V, Shi Xianglin, et al. New insights into the role of nuclear factor-?B,a Ubiquitous transcription factor in the initiation of diseases. Clinical Chemistry, 1999, 45(1):7-17
69 Nishimune H,Vasseur S,Wiese S,et al.Reg2 is a motoneuron neurotrophic factor and a singalling intermediate in the CNTF survival pathway.Nat Cell Biol,2000,2(12):906-914
70 Hietarranta A,Mustonnen H,Puolakkainen P,et al. Pro- inflammatory effects of pancreatic elastase are mediated through TLR4 and NF-?appaB.Biochem Biophys Res Commun, 2004, 323(1):192-196
71刘洪斌,崔乃强,李东华,等.磷脂酶A2与NF-?B在大鼠急性坏死性胰腺炎肺损伤中的作用.中国中西医结合外科杂志,2005,11(2):115-119