PI3K/PKB信号通路及其抑制剂wortmannin在大鼠急性胰腺炎相关肠道损伤中的作用
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摘要
急性胰腺炎(acute pancreatitis, AP)是临床上常见的急腹症之一,常表现为起病急、变化快、病情重、除发生胰腺病变外,常伴有多器官功能损伤,肠道功能损伤是急性胰腺炎的重要并发症之一。肠粘膜屏障受损时,大量内毒素、细菌释放入血,引起内毒素血症或菌血症,加重全身炎症反应,甚至引起多器官功能衰竭。因此,积极预防和治疗肠道损伤已经成为急性胰腺炎病情转归的重要影响因素。肠道损伤主要表现为动力障碍,肠管损伤和粘膜屏障功能损害三方面,目前认为其机制主要与肠管炎症细胞的浸润,肿瘤坏死因子(TNF-α)等细胞因子大量释放,肠腔细菌过度繁殖及过量氧自由基产生有关。磷脂酰肌醇-3激酶(phosphatidylinositol 3-kinase,PI3K)是一种与细胞内信号转导有关的第二信使,蛋白激酶B(protein kinase B,PKB)是PI3K的一个下游作用因子,PI3K/PKB通路是一个重要的信号调控系统,可介导细胞外信号引起细胞反应,在炎症细胞的活化、趋化、肿瘤的发生、缺血再灌注及间质纤维化等方面发挥了重要作用。近年来PI3K/PKB信号通路与胰腺炎的关系受到越来越多的重视。急性胰腺炎时,PI3K/PKB信号通路被激活,进一步激活下游作用因子NF-κB,促进多种炎症因子的产生,进而活化并募集中性粒细胞产生更多细胞因子与炎症介质,引起级联放大效应,即所谓的“炎症因子风暴”,诱发全身炎症反应,甚至多器官功能衰竭。通常情况下p-PKB可衡量PI3K/PKB信号通路的活化程度。本实验通过测定肠道组织中P-PKB及PKB的表达水平,进而研究PI3K/PKB信号通路与大鼠AP相关肠道损伤的关系,同时应用PI3K信号通路抑制剂wortmannin干预AP大鼠,观察其在AP大鼠并发肠道损伤的作用并研究其可能的机制。
目的:通过应用PI3K特异性抑制剂wortmannin预处理大鼠AP模型,探讨wortmannin在急性胰腺炎相关肠道损伤中的作用,并研究其可能的作用机制。
方法:将健康成年SD大鼠54只随机分为对照组(sham operation SO)、急性胰腺炎(acute pancreatitis AP)组和AP+wortmannin(WAP)组,每组18只,每组又以不同时间点3h、6h、12h分为三个亚组,每个亚组6只大鼠。以10%的水合氯醛腹腔注射麻醉,取上腹正中切口进入腹腔,以无创动脉夹夹闭近肝门端胆管,然后用加工钝头1ml注射器于十二指肠乳头附近逆行插入胰胆管,以0.2 ml/min速度逆行胆胰管注射50g/L牛磺胆酸钠制备AP模型,推药后以左手拇指捏闭胰胆管入十二指肠处,观察3 min,去除动脉夹,其中WAP组于造模前4小时腹腔注射wortmannin(0.35mg/kg)。分别于造模后3h、6h、12h每组处死6只大鼠,下腔静脉取血,检测血清淀粉酶、TNF-α(tumour necrosis factor-α)水平、血浆内毒素的水平,观察胰腺和肠道的大体及病理学改变,并检测肠道组织髓过氧化物酶(myeloperoxidase, MPO)活性,采用western blot检测肠道组织中PKB及P-PKB的表达水平,观察PI3K/PKB信号通路与大鼠AP相关肠道损伤的关系,及应用wortmannnin预处理AP大鼠后,上述测量指标的变化。
结果:SO组的各测量指标在各时间点无明显变化,与SO组相比,AP组在各时间点血清淀粉酶、TNF-α水平、血浆内毒素及肠道组织MPO活性均显著升高(P<0.05),胰腺、肠道病理损伤亦较SO组明显加重,中性粒细胞浸润增多;WAP组在各时间点较SO组各项指标均升高,但与AP组相比均明显降低(P<0.05),肠道组织中PKB的表达在三个组比较没有明显差别,P-PKB的表达为:AP组和WAP组均较正常对照组明显升高,WAP组较AP组比较明显降低。
结论:PI3K\PKB信号通路参与了AP大鼠肠道损伤,其抑制剂wortmannin有一定的保护作用,机制可能与其抑制了中性粒细胞的浸润,多种炎症细胞的活化和TNF-α等炎症因子的释放受到抑制有关。
Acute pancreatitis (AP) is one of the most frequent clinical acute abdominal, in addition to pancreatic lesions, often complicated with multiple-organ dysfunction syndrome(MODS), intestinal injury is one of the most important complications. When intestinal mucosa damage, lots of endotoxin, bacteria are released into the blood, cause endotoxemia or bacteremia, aggravating systemic inflammation, even causing MODS. Therefore, the actively prevention and treatment of intestinal damage has become important influence factors of AP. Intestinal injury mainly performance for dyskinesis, the damage of dilated bowel and mucous membrane barrier function, the mechanism mainly with inflammatory cells infiltrating loops, the releasing of inflammatory mediators-tumor necrosis factor (TNF alpha)and cytokines, oxygen-free radicals and bacteria excessive breeding in intestinal canal. Phosphatidylinositol 3-kinase(PI3K) is a second messenger that is related to intracellular signal transduction. Protein kinase B(PKB) is a downstream effector of PI3K. PI3K∕PKB signal pathway is an important signal system which causes celluar response through mediating extracelluar signaling. It plays important roles in the following fields: activation and chemotaxis of inflammatory factors, cancer progression, ischemia-reperfusion, interstitial fibrosis and so on. Recently, the relationship between PI3K∕PKB signal pathway and pancreatitis obtain more and more attention. When PI3K∕PKB signal pathway is activated in pancreatitis, its products can activate the downstream factors. These activated downstream factors can recruit neutrophil to produce inflammatory factors and lead to cascaded amplification, which called“cytokine storm”. Mutiple organs are damaged in the reactions. By increasing the intra-cellular Ca2+ level, PI3K can activate trypsinogen and promote the development of pancreatitis. Normally, phosphorylated P-PKB can measure the activation of PI3K∕PKB signal pathway. In our test, the expressional levels of P-PKB and PKB in intestine were measured ,which is used to study the relationship between PI3K∕PKB signal pathway and intestinal injuries induced by AP in rat. Our study use the specific inhibitor of PI3K wortmannin to interfere in intestinal injuries induced by AP and investigate its possible mechanism.
Objective: To investigate the role of wortmannin in intestinal injuries induced by acute pancreatitis in rats and its mechanism.
Methods: Fifty-four healthy male SD rats were randomly divided into 3 groups: SO group(sham operation group), AP group (acute pancreatitis group),WAP group(acute pancreatitis plus wortmannin).(n=18 per group). AP modle was induced by retrograde injection of 50g/L sodium taurocholate into the biliopancreatic duct of rats. wortmannin was administered to WAP group introperitoneally with the dosage of 0.35mg/kg 4 hours before the modle was made. The serum levels of amylase, tumor necrosis factor-alpha ( TNF-α), endotoxin were measured; myeloperoxidase (MPO) activity in intestine was minitored; the expressional levels of PKB and P- PKB were measured by western blot; intestine and pancreas were collected to examine pathological changes.
Results: The serum levels of tumor necrosis factor-alpha ( TNF-α), amylase, endotoxin and myeloperoxidase (MPO) activity in intestine of AP group were significantly elevated ( P < 0. 05) compared with SO group. The pathological changes in intestine and pancreas were worsened as time prolonging; all indices in WAP group were significantly elevated ( P < 0. 05) compared with SO group and significantly decreased ( P < 0. 05) compared with AP group. There is no significant difference of PKB expression among the 3 groups. The P-PKB expression in AP group and WAP group was significantly elevated ( P < 0. 05) compared with SO group, and the P-PKB expression in WAP group was significantly decreased ( P < 0. 05) compared with AP group .
Conclusions: PI3K∕PKB signal transduction pathway is involved in the pathogenesis of intestine injuries induced by acute pancreatitis in rats, pretreatment with wortmannin can inhibit the PI3K∕PKB signal transduction pathway, thus relieving the intestinal injuries induced by acute pancreatitis. The mechanism is related to the inhibition of activation and chemotaxis of inflammatory factors and neutrophil aggregation.
目的:通过应用PI3K特异性抑制剂wortmannin预处理大鼠AP模型,探讨wortmannin在急性胰腺炎相关肠道损伤中的作用,并研究其可能的作用机制。
方法:将健康成年SD大鼠54只随机分为对照组(sham operation SO)、急性胰腺炎(acute pancreatitis AP)组和AP+wortmannin(WAP)组,每组18只,每组又以不同时间点3h、6h、12h分为三个亚组,每个亚组6只大鼠。以10%的水合氯醛腹腔注射麻醉,取上腹正中切口进入腹腔,以无创动脉夹夹闭近肝门端胆管,然后用加工钝头1ml注射器于十二指肠乳头附近逆行插入胰胆管,以0.2 ml/min速度逆行胆胰管注射50g/L牛磺胆酸钠制备AP模型,推药后以左手拇指捏闭胰胆管入十二指肠处,观察3 min,去除动脉夹,其中WAP组于造模前4小时腹腔注射wortmannin(0.35mg/kg)。分别于造模后3h、6h、12h每组处死6只大鼠,下腔静脉取血,检测血清淀粉酶、TNF-α(tumour necrosis factor-α)水平、血浆内毒素的水平,观察胰腺和肠道的大体及病理学改变,并检测肠道组织髓过氧化物酶(myeloperoxidase, MPO)活性,采用western blot检测肠道组织中PKB及P-PKB的表达水平,观察PI3K/PKB信号通路与大鼠AP相关肠道损伤的关系,及应用wortmannnin预处理AP大鼠后,上述测量指标的变化。
结果:SO组的各测量指标在各时间点无明显变化,与SO组相比,AP组在各时间点血清淀粉酶、TNF-α水平、血浆内毒素及肠道组织MPO活性均显著升高(P<0.05),胰腺、肠道病理损伤亦较SO组明显加重,中性粒细胞浸润增多;WAP组在各时间点较SO组各项指标均升高,但与AP组相比均明显降低(P<0.05),肠道组织中PKB的表达在三个组比较没有明显差别,P-PKB的表达为:AP组和WAP组均较正常对照组明显升高,WAP组较AP组比较明显降低。
结论:PI3K\PKB信号通路参与了AP大鼠肠道损伤,其抑制剂wortmannin有一定的保护作用,机制可能与其抑制了中性粒细胞的浸润,多种炎症细胞的活化和TNF-α等炎症因子的释放受到抑制有关。
Acute pancreatitis (AP) is one of the most frequent clinical acute abdominal, in addition to pancreatic lesions, often complicated with multiple-organ dysfunction syndrome(MODS), intestinal injury is one of the most important complications. When intestinal mucosa damage, lots of endotoxin, bacteria are released into the blood, cause endotoxemia or bacteremia, aggravating systemic inflammation, even causing MODS. Therefore, the actively prevention and treatment of intestinal damage has become important influence factors of AP. Intestinal injury mainly performance for dyskinesis, the damage of dilated bowel and mucous membrane barrier function, the mechanism mainly with inflammatory cells infiltrating loops, the releasing of inflammatory mediators-tumor necrosis factor (TNF alpha)and cytokines, oxygen-free radicals and bacteria excessive breeding in intestinal canal. Phosphatidylinositol 3-kinase(PI3K) is a second messenger that is related to intracellular signal transduction. Protein kinase B(PKB) is a downstream effector of PI3K. PI3K∕PKB signal pathway is an important signal system which causes celluar response through mediating extracelluar signaling. It plays important roles in the following fields: activation and chemotaxis of inflammatory factors, cancer progression, ischemia-reperfusion, interstitial fibrosis and so on. Recently, the relationship between PI3K∕PKB signal pathway and pancreatitis obtain more and more attention. When PI3K∕PKB signal pathway is activated in pancreatitis, its products can activate the downstream factors. These activated downstream factors can recruit neutrophil to produce inflammatory factors and lead to cascaded amplification, which called“cytokine storm”. Mutiple organs are damaged in the reactions. By increasing the intra-cellular Ca2+ level, PI3K can activate trypsinogen and promote the development of pancreatitis. Normally, phosphorylated P-PKB can measure the activation of PI3K∕PKB signal pathway. In our test, the expressional levels of P-PKB and PKB in intestine were measured ,which is used to study the relationship between PI3K∕PKB signal pathway and intestinal injuries induced by AP in rat. Our study use the specific inhibitor of PI3K wortmannin to interfere in intestinal injuries induced by AP and investigate its possible mechanism.
Objective: To investigate the role of wortmannin in intestinal injuries induced by acute pancreatitis in rats and its mechanism.
Methods: Fifty-four healthy male SD rats were randomly divided into 3 groups: SO group(sham operation group), AP group (acute pancreatitis group),WAP group(acute pancreatitis plus wortmannin).(n=18 per group). AP modle was induced by retrograde injection of 50g/L sodium taurocholate into the biliopancreatic duct of rats. wortmannin was administered to WAP group introperitoneally with the dosage of 0.35mg/kg 4 hours before the modle was made. The serum levels of amylase, tumor necrosis factor-alpha ( TNF-α), endotoxin were measured; myeloperoxidase (MPO) activity in intestine was minitored; the expressional levels of PKB and P- PKB were measured by western blot; intestine and pancreas were collected to examine pathological changes.
Results: The serum levels of tumor necrosis factor-alpha ( TNF-α), amylase, endotoxin and myeloperoxidase (MPO) activity in intestine of AP group were significantly elevated ( P < 0. 05) compared with SO group. The pathological changes in intestine and pancreas were worsened as time prolonging; all indices in WAP group were significantly elevated ( P < 0. 05) compared with SO group and significantly decreased ( P < 0. 05) compared with AP group. There is no significant difference of PKB expression among the 3 groups. The P-PKB expression in AP group and WAP group was significantly elevated ( P < 0. 05) compared with SO group, and the P-PKB expression in WAP group was significantly decreased ( P < 0. 05) compared with AP group .
Conclusions: PI3K∕PKB signal transduction pathway is involved in the pathogenesis of intestine injuries induced by acute pancreatitis in rats, pretreatment with wortmannin can inhibit the PI3K∕PKB signal transduction pathway, thus relieving the intestinal injuries induced by acute pancreatitis. The mechanism is related to the inhibition of activation and chemotaxis of inflammatory factors and neutrophil aggregation.
引文
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25 Branger J, vanden Blink B, W eijer S, et al. Anti-inflammatory effects of a p38 mitogen activated proteinkinase inhibitor during human endotoxemia. J Immunol. 2002, 168(8):4070-4077
26 Wager AC, Metzler W, Hofer T, et al. p38MAP kinase is expressed in the pancreas and is immediately activated following cerulean hyperstimulation. Digestion, 1999, 60(1):41-47
27 Blinman, Ilya Gukovsky, Michelle Mouria, et al. Activation of pancreatic acinar cells on isolation from tissue cytokine up regulation via p38MAP kinase. Am J Physiol Cell Physiol, 2000, 279(6):1993-2003
28任洪波,许国铭,李兆申,等.重症急性胰腺炎大鼠丝裂原活化蛋白激酶信号转导通路的动态变化.中华消化杂志, 2003, 23(9):548-550。
29任洪波,李延青,程宝泉,等.丝裂原活化蛋白激酶信号转导通路抑制剂对重症急性胰腺炎治疗作用的实验研究.中华消化杂志, 2006, 26(5):343-345
30任洪波,许国铭,李兆申,等. CNI-1493对重症急性胰腺炎大鼠外周血多形核粒细胞功能的影响.中国胰腺病杂志, 2003, 3(1):23-25
31 Fleischer F, Dabew R, Goke B, et al. Stress kinase inhibition modulates acute experimental pancreatitis.World J Gastroenterol, 2001, 7(2):259
32 Dabrowski A. Exocrine pancreas; molecular basis for intracellular signaling, damage and protection-Polish experience. J Physiol Pharmacol, 2003, 54(Suppl 3):167-181
33 Chen P, Huang L, Zhang Y, et al. The antagonist of the JAK-1/STAT-1 signaling pathway improves the severity of cerulein-stimulated pancreatic injury via inhibition of NF-κB activity. Int J Mol Med, 2011, 27(5):731-738.
34 Minutoli L, Altavilla D, Marini H, et al. Protective effects of SP600125 anew inhibitor of c-jun N-terminal kinase (JNK) and extracellular-regulated kinase (ERK1/2) in an experimental model of cerulein-induced pancreatitis. Life Sci, 2004, 75(24):2853-2866
35 Imitrova P, Gyurkovska V, Shalova I, et al. Inhibition of zymosan-induced kidney dysfunction by tyrphostin AG-490. J Inflamm, 2009, 5:6-13.
36张勉之,赵松,段建召,等. JAK/STAT信号转导途径在糖尿病肾病肾小管上皮细胞中的表达.中国中西医结合急救杂志, 2007, (5):309-312
37 Kovacic JC, Gupta R, Lee AC, et al. Stat3-dependent acute Rantes production in vascular smooth muscle cells modulates inflammation following arterial injury in mice. J Clin Invest, 2010, 120(1):303-314
38 Xiong H, Zhang ZG, Tian XQ, et al. Inhibition of JAK1, 2/STAT3signaling induces apoptosis, cell cycle arrest, and reduces tumor cell invasion in colorectal cancer cells. Neoplasia, 2008, 10:287-297
39 Lassmann S, Schuster I, Walch A, et al. STAT3mRNA and protein expression in colorectal cancer: effects on STAT3-inducible targets linked to cell survival and proliferation. J ClinPathol, 2007, 60:173-179
40 Zhang X, Darnell JE Jr. Functional importance of Stat3 tetramerization in activation of the alpha 2-macroglobulingene. J Biol Chem, 2001, 276 (36): 33576-33581
41 Lee C, Lim HK, Sakong J, et al. Janus kinase-signal transducer and activator of transcription mediates phosphatidic acid-induced interleukin (IL)-1beta and IL-6 production. Mol Pharmacol, 2006, 69 (3):1041-1047
42 Horvath CM.The Jak-STAT pathway stimulated by interleukin 6. Sci STKE, 2004(260):tr9
43钟敦璟,郭红,郝嘉,等. STAT3对大鼠重症急性胰腺炎血清体外作用AT-ⅡSP-C的影响.第三军医大学学报, 2008, 30(19):1807-1809
44郭婧芸,朱人敏,张晓华,等.抑制JAK/STAT信号通路对实验性急性胰腺炎大鼠肝损伤的保护作用.胃肠病学, 2008, 13(12): 719-722
45周静,王杉,安友仲,等. STAT3在急性重症胰腺炎大鼠肠黏膜屏障功能障碍中的作用.中华普通外科杂志, 2005, 20(3):167-170
46刘刚,周朝霞,姬新才,等.急性胰腺炎患者血浆D-二聚体含量及外周血淋巴细胞蛋白激酶C活性的变化及意义.中国急救医学, 2005, 25 (18): 550-553
47 Horovitz FriedM, Sampson SR. Involvement of PKC alpha in insulin-induced PKC delta expression: Importance of SP1 and NFkappaB transcription factors. Biochem Biophys Res Commun, 2007, 352(1):78-83
48 Zhang L , Zhang X, westlund KN. Restoration of spontaneous exploratory behaviors with an intrathecal NMDA receptor antagonistor a PKC inhibitor in rats with acute pancreatitis. Phar maco l Biochem Behav, 2004, 77(1):145-153
49 Frossi B, vera J, rsch E, Pucillo C. Selective activation of Fyn/PI3K and P38MAPK regulates IL-4 Production in BMMC under nontoxic stress condition. J Immunol, 2007, 178(4):2549-2555
50 Steelman LS, Pohnert S, Shelton JG, et al. JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis. Leukemia, 2004, 18(2):189-218
51 F leischer F, Dabew R, Goke B, et al. Stress kinase inhibition modulates acute experimental pancreatitis. World J Gastroenterol, 2001, 7(2):259-265
52 Karim A, Derek S, Sanchayita M, et al. Involvement of PKCa/h in TLR4 and TLR2 dependent activation of NF-κB. Cellular Signalling, 2005, 17 (3):385-394
53 Comalada MJ, Xaus AF, Valledor C, et al. PKC is involved in JNK activation that mediates LPS induced TNF-αwhich induces apoptosis in macrophages. Am J Cell Physiol, 2003, 285(5):C1235-C1245
2 Wang XP, Wang BX, Wu K, et al. Excessive cytokine expression mediated by NF-κB of intestinal mucosa and role of somatropin in acute necrotic pancreatitis. Chin J Hepatobiliary Surg, 2003, 9(1):45-49
3 Mole DJ, Taylor MA, McFerran NV, et al. The isolated perfused liver response to a‘second hit’of portal endotoxin during severe acute pancreatitis. Pancreatology, 2005, 5(4-5):475-485
4 Li Z, Jiang H, Xie W, et al. Role of PLC-β2 and -β3and PI3Kγin chemoattractant-mediated signal transduction. Science, 2000, 287(5455): 1046-1049
5 Yum HK, Arcaroli J, Kupfner J, et al. Involvement of phosphoinositide 3 -kinases in neutrophil activation and the development of acute lung injury. J Immunol, 2001, 167(11): 6601-6608
6 Uhlig U, Fehrenbach H, Lachmann RA, et al. Phosphoinositide 3-OH kinase inhibition prevents ventilation -induced lung cell activation. Am J Respir Crit Care Med, 2004, 169(2):201-208
7 Wei J, Feng J. Signaling pathways associated with inflammatory bowel disease. Recent Pat Inflamm Allergy Drug Discov. 2010, Jun 1,4(2):105-107
8 Aho HJ, Koskensalo SM, Nevalainen TJ. Experimental pancreatitis in the rat. Sodium taurocholate-induced acute haemorrhagic pancreatitis. Scand J Gastroenterol, 1980, 15(4):411-416
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10 Singh VP, Saluja AK, Bhagat L, et al. Phosphatidylinosito3-kinase dependent activation of tryp sinogen modulates the severity of acutepancreatitis. J Clin Invest, 2001, 108(9):1387-1395
11 Surbatovic M, Jovanovic K, Radakovi? S, et al. Pathophysiological aspects of severe acute pancreatitis-associated lung injury. Srp Arh Celok Lek, 2005, 133(1-2):76-81
12 Zhang XP, Li ZJ and Zhang J. Inflammatory mediators and microcirculatory disturbance in acute pancreatitis. Hepatobiliary Pancreat Dis Int, 2009, 8(4):351-357
13杨元生,崔淑兰,陈垦,等.重症急性胰腺炎实验动物模型的研究进展.世界华人消化杂志, 2009, 17(25):2601-2606
14 Wang XP, Wang BX, Wu K, et al. Excessive cytokine expression mediated by NF-κB of intestinal mucosa and role of somatropin in acute necrotic pancreatitis. Chin J Hepatobiliary Surg, 2003, 9(1):45-49
15 Gustin JA, Ozes ON, Akca H, et al. Cell type-specific expression of the I kappa B kinases determines the significance of phosphatidylinositol 3-kinase /Akt signaling to NF-kappaB activation. J Biol Chem, 2004, 279 (3):1615-1620
16 Yum HK, Arcaroli J, Kupfner J, et al. Involvement of Phosphoinositide-3-Kinases in Neutrophil Activation and the Development of Acute Lung Injury. J Immunol, 2001, 167(11):6601-6608
17 Makhijia R, Kingsnorth AN. Cytokine storm in acute pancreatitis. Hepatobiliary Pancreat Surg, 2002, 9(4):401-410
18 Anne Demols, Jacques Deviere. New Frontiers in the Pharmacological Precention of Post-ERCP Pancreatitis:The Cytokines JOP. Pancreas, 2003, 4(1):49-57
19 Li Z, Jiang H, Zie W, et al. Role of PLC-β2 and -β3 and PI3Kγin chemoattractant-mediated signal transduction. Science, 2000, 287(5455):1046-1049
20 Hirsch E, Katanaev VL, Garlanda C, et al. Central role for G protein-coupled phosphoinositide 3-kinase gamma in inflammation. Science, 2000, 287(5455):1049-1053
21 Wipf P, Halter RJ. Chemistry and biology of wortmannin. Org BiomolChem, 2005, 3(11):2053-2061
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23张成,栾正刚,郭仁宣,等. PI3K/PKB信号转导通路在重症急性胰腺炎胰腺损伤中的作用.中国普通外科杂志, 2007, 16(10):968-971
24 Yang SJ, Chen HM, Hsieh CH, et al. Akt pathway is required for oestrogen-mediated attenuation of lung injury in a rodent model of cerulein-induced acute pancreatitis. Injury, 2011, 42(7):638-642
25 Fischer L, Gukovskaya AS, Penninger JM, et al. Phosphatidylinositol 3-kinase facilitates bile acid-induced Ca2+ responses in pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol, 2007, 292(3):G875-G886
26 Saluja AK, Bliagat L, Lee HS, et al. Secretagogue-induced digestive enzyme activation and cell injury in rat pancreatic acini. Am J Physiol, 1999, 276(4 pt 1):G836-G842
27 Olanders K,Borjesson A,Zhao X,et al.Effects of anticoagulant treatment on intestinal ischaemia and reperfusion injury in rats. Acta Anaesthesiol Scand, 2005, 49(4):517-524
1孟艳,米蕊芳,赵春华. PI3K-AKT-mTOR信号通路在细胞分化中的作用.基础医学与临床, 2007, 27(12):1404-1408
2郭琳,王强. PI3K/Akt/mTOR信号传导通路与恶性肿瘤浸润和转移的研究进展.现代肿瘤医学, 2009, 17 (08):1585-1589
3 Ma J, Sawai H, Ochi N, et al. PTEN regulates angiogenesis through PI3K/Akt/VEGF signaling pathway in human pancreatic cancer cells. Mol Cell Biochem, 2009,331(1-2):161-171
4 Edling CE, Selvaggi F, Buus R, et al. Key role of phosphoinositide 3-kinase class IB in pancreatic cancer. Clin Cancer Res, 2010, 16(20):4928-4937
5 Frojdo S, Durand C, Molin L, et al. Phosphoinositide 3-kinase as a novel functional target for the regulation of the insulin signaling pathway by SIRT1, 2011, 335(2):166-176
6 Manukyan MC, Weil BR, Wang Y, et al. The phosphoinositide-3 kinase survival signaling mechanism in sepsis. Shock, 2010, 34(5):442-449
7 Uhlig U, Fehrenbach H, Lachmann RA, et al. Phosphoinositide3-OH kinase inhibition prevents ventilation-induced lung cell activation. Am J Resp ir Crit Care Med, 2004, 169(2):201-208
8 Wei J, Feng J. Signaling pathways associated with inflammatory bowel disease. Recent Pat Inflamm Allergy Drug Discov, 2010, 4(2):105-107
9何忠野,郭仁宣,谢成耀,等.硫酸软骨素对ANP大鼠胰腺细胞氧化应激损伤及细胞间连接的影响.中国普通外科杂志, 2006, 15(8):590-594
10张翼,吕新生,孙维佳,等.甘遂对重症急性胰腺炎大鼠核因子2κB活化的影响.中国普通外科杂志, 2006, 15(10):761-765
11张成,栾正刚,郭仁宣,等. PI3K/PKB信号转导通路在重症急性胰腺炎胰腺损伤中的作用.中国普通外科杂志, 2007, 16(10):968-971
12 Lupia E, Goffi A, De Giuli P, et al. Ablation of phosphoinositide-3-kinase -gamma reduces the severity of acute pancreatitis. Am J Pathol, 2004, 165(6):2003-2011
13李丙所,蔡文科,任彦顺,等. Wortmannin对重症急性胰腺炎大鼠胰肝损伤的保护作用.中国普通外科杂志, 2007, 16(10):976-978
14 Gukovsky I, Cheng JH , Nam KJ, et al. Phosphatidylinositide3-kinase gamma regulates key pathologic responses to chole-cystokinin in pancreatic acinar cells. Gastroenterology, 2004, 126(2):554-566
15 Petersen OH. Ca2+ induced pancreatic cell death:roles of the endoplasmic reticulum, zymogen granules, lysosomes and endosomes. Gastroenterol Hepatol, 2008, 23(Suppl 1):S31- S36
16黄芬,陈新宙.重症急性胰腺炎发病机制.实用临床医学, 2006, 7(10): 204-207
17 Fischer L, Gukovskaya AS, Penninger JM, et al. Phosphatidylinositol 3-kinase facilitates bile acid-induced Ca2+ responses in pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol, 2007, 292(3):G875-G886
18 Xi-Ping Zhang, Zhi-Jun Li and Jie Zhang. Inflammatory mediators and microcirculatory disturbance in acute pancreatitis. Hepatobiliary Pancreat Dis Int, 2009, 8(4): 351-357
19 Gustin JA, Ozes ON, Akca H, et al. Cell type-specific expression of the IkappaB kinases determines the significance of phosphatidylinositol 3-kinase /Akt signaling to NF-kappaB activation. J Biol Chem, 2004, 279 (3):1615-1620
20李丙所,蔡文科,任彦顺,等. Wortmannin对重症急性胰腺炎大鼠胰肝损伤的保护作用.中国普通外科杂志, 2007, 16(10):976-978
21 LuPia E, Goffi A, DeGiulip, et al. Ablation of phos-Phoinositide3-kinase-gamma reduces the severity of acute pancreatitis. Am J Pnthof, 2004, 165(6):2003-2011
22 Larmonier CB, Midura-Kiela MT, Ramalingam R, et al. Modulation of neutrophil motility by curcumin: Implications for inflammatory bowel disease. Inflamm Bowel Dis, 2011, 17(2):503-515
23张成,奕正刚,葛春林,等.磷脂酞肌醇3激酶/蛋白激酶B信号转导通路在重症急性胰腺炎中性粒细胞活化中的作用.世界华人消化杂志, 2006, 14(20):1987-1991。
24 Olson CM, Hedrick MN, Bates HI, et al. P38 Mitogen- activated protein kinase controls NF-κB transcriptional activation and tumor necrosis factor alpha production through RelA phosphorylation mediated by mitogen- and stress- activated protein kinase 1 in response to borrelia burgdorferi antigens. Interfection and Immunity, 2007, 75(1):270-277
25 Branger J, vanden Blink B, W eijer S, et al. Anti-inflammatory effects of a p38 mitogen activated proteinkinase inhibitor during human endotoxemia. J Immunol. 2002, 168(8):4070-4077
26 Wager AC, Metzler W, Hofer T, et al. p38MAP kinase is expressed in the pancreas and is immediately activated following cerulean hyperstimulation. Digestion, 1999, 60(1):41-47
27 Blinman, Ilya Gukovsky, Michelle Mouria, et al. Activation of pancreatic acinar cells on isolation from tissue cytokine up regulation via p38MAP kinase. Am J Physiol Cell Physiol, 2000, 279(6):1993-2003
28任洪波,许国铭,李兆申,等.重症急性胰腺炎大鼠丝裂原活化蛋白激酶信号转导通路的动态变化.中华消化杂志, 2003, 23(9):548-550。
29任洪波,李延青,程宝泉,等.丝裂原活化蛋白激酶信号转导通路抑制剂对重症急性胰腺炎治疗作用的实验研究.中华消化杂志, 2006, 26(5):343-345
30任洪波,许国铭,李兆申,等. CNI-1493对重症急性胰腺炎大鼠外周血多形核粒细胞功能的影响.中国胰腺病杂志, 2003, 3(1):23-25
31 Fleischer F, Dabew R, Goke B, et al. Stress kinase inhibition modulates acute experimental pancreatitis.World J Gastroenterol, 2001, 7(2):259
32 Dabrowski A. Exocrine pancreas; molecular basis for intracellular signaling, damage and protection-Polish experience. J Physiol Pharmacol, 2003, 54(Suppl 3):167-181
33 Chen P, Huang L, Zhang Y, et al. The antagonist of the JAK-1/STAT-1 signaling pathway improves the severity of cerulein-stimulated pancreatic injury via inhibition of NF-κB activity. Int J Mol Med, 2011, 27(5):731-738.
34 Minutoli L, Altavilla D, Marini H, et al. Protective effects of SP600125 anew inhibitor of c-jun N-terminal kinase (JNK) and extracellular-regulated kinase (ERK1/2) in an experimental model of cerulein-induced pancreatitis. Life Sci, 2004, 75(24):2853-2866
35 Imitrova P, Gyurkovska V, Shalova I, et al. Inhibition of zymosan-induced kidney dysfunction by tyrphostin AG-490. J Inflamm, 2009, 5:6-13.
36张勉之,赵松,段建召,等. JAK/STAT信号转导途径在糖尿病肾病肾小管上皮细胞中的表达.中国中西医结合急救杂志, 2007, (5):309-312
37 Kovacic JC, Gupta R, Lee AC, et al. Stat3-dependent acute Rantes production in vascular smooth muscle cells modulates inflammation following arterial injury in mice. J Clin Invest, 2010, 120(1):303-314
38 Xiong H, Zhang ZG, Tian XQ, et al. Inhibition of JAK1, 2/STAT3signaling induces apoptosis, cell cycle arrest, and reduces tumor cell invasion in colorectal cancer cells. Neoplasia, 2008, 10:287-297
39 Lassmann S, Schuster I, Walch A, et al. STAT3mRNA and protein expression in colorectal cancer: effects on STAT3-inducible targets linked to cell survival and proliferation. J ClinPathol, 2007, 60:173-179
40 Zhang X, Darnell JE Jr. Functional importance of Stat3 tetramerization in activation of the alpha 2-macroglobulingene. J Biol Chem, 2001, 276 (36): 33576-33581
41 Lee C, Lim HK, Sakong J, et al. Janus kinase-signal transducer and activator of transcription mediates phosphatidic acid-induced interleukin (IL)-1beta and IL-6 production. Mol Pharmacol, 2006, 69 (3):1041-1047
42 Horvath CM.The Jak-STAT pathway stimulated by interleukin 6. Sci STKE, 2004(260):tr9
43钟敦璟,郭红,郝嘉,等. STAT3对大鼠重症急性胰腺炎血清体外作用AT-ⅡSP-C的影响.第三军医大学学报, 2008, 30(19):1807-1809
44郭婧芸,朱人敏,张晓华,等.抑制JAK/STAT信号通路对实验性急性胰腺炎大鼠肝损伤的保护作用.胃肠病学, 2008, 13(12): 719-722
45周静,王杉,安友仲,等. STAT3在急性重症胰腺炎大鼠肠黏膜屏障功能障碍中的作用.中华普通外科杂志, 2005, 20(3):167-170
46刘刚,周朝霞,姬新才,等.急性胰腺炎患者血浆D-二聚体含量及外周血淋巴细胞蛋白激酶C活性的变化及意义.中国急救医学, 2005, 25 (18): 550-553
47 Horovitz FriedM, Sampson SR. Involvement of PKC alpha in insulin-induced PKC delta expression: Importance of SP1 and NFkappaB transcription factors. Biochem Biophys Res Commun, 2007, 352(1):78-83
48 Zhang L , Zhang X, westlund KN. Restoration of spontaneous exploratory behaviors with an intrathecal NMDA receptor antagonistor a PKC inhibitor in rats with acute pancreatitis. Phar maco l Biochem Behav, 2004, 77(1):145-153
49 Frossi B, vera J, rsch E, Pucillo C. Selective activation of Fyn/PI3K and P38MAPK regulates IL-4 Production in BMMC under nontoxic stress condition. J Immunol, 2007, 178(4):2549-2555
50 Steelman LS, Pohnert S, Shelton JG, et al. JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis. Leukemia, 2004, 18(2):189-218
51 F leischer F, Dabew R, Goke B, et al. Stress kinase inhibition modulates acute experimental pancreatitis. World J Gastroenterol, 2001, 7(2):259-265
52 Karim A, Derek S, Sanchayita M, et al. Involvement of PKCa/h in TLR4 and TLR2 dependent activation of NF-κB. Cellular Signalling, 2005, 17 (3):385-394
53 Comalada MJ, Xaus AF, Valledor C, et al. PKC is involved in JNK activation that mediates LPS induced TNF-αwhich induces apoptosis in macrophages. Am J Cell Physiol, 2003, 285(5):C1235-C1245