大鼠重症急性胰腺炎发病机制中p38丝裂原活化蛋白激酶的作用
施新岗, 李兆申, 贾一韬, 许永春, 满晓华, 龚燕芳, 屠振兴, 许国铭. 中国人民解放军第二军医大学附属长海医院消化内科 上海市 200433
施新岗,男, 1970-02-22生, 江苏省启东市人, 汉族, 2004年第二军医大学博士, 主治医师, 主要从事重症急性胰腺炎发病机制和治疗研究
项目负责人: 李兆申，200433，上海市长海路174号，中国人民解放军第二军医大学长海医院消化内科. zhsli@81890.net
电话: 021-25070585
收稿日期: 2004-08-17 接受日期: 2004-10-11
, http://www.100md.com
p38 mitogen-activated protein kinase signal transduction pathway in pathogenesis of severe acute pancreatitis in rats
Xin-Gang Shi, Zhao-Shen Li, Yi-Tao Jia, Yong-Chun Xu, Xiao-Hua Man, Yan-Fang Gong, Zhen-Xing Tu, Guo-Ming Xu
Xin-Gang Shi, Zhao-Shen Li, Yi-Tao Jia, Yong-Chun Xu, Xiao-Hua Man, Yan-Fang Gong, Zhen-Xing Tu, Guo-Ming Xu, Department of Gastroenterology of Changhai Hospital, Second Military Medical University, Shanghai 200433, China
, 百拇医药
Correspondence to: Zhao-Shen Li, Department of Gastroenterology, Changhai Hospital, Second Military Medical University, 174 Changhai Lu, Shanghai 200433, China. zhsli@81890.net
Received: 2004-08-17 Accepted: 2004-10-11
Abstract
AIM: To study the dynamic changes of p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation and to assess the effect of p38 MAPK phosphorylation inhibitor in severe acute pancreatitis (SAP) in rats.
, 百拇医药
METHODS: The SAP model was induced by bili-pancreatic duct retrograde infusion with 5% sterile sodium taurocholate solution. Eighty Sprague-Dawley (SD) rats were randomly divided into sham-operation (SO) group (n = 30), SAP-NS group (n = 25) and SAP-CNI1493 group (n = 25). In the SAP-CNI1493 group, SD rats were administered with 10 mg/kg inhibitor CNI-1493 (i.v.) 30 min before induction of SAP. In SAP-NS group, rats received same volume isotonic saline (i.v.) as CNI-1493. Pancreatic tissues and serum samples were collected before and 15 min, 0.5 h, 1 h, 3 h, 6 h after operation. Western blot analysis was performed to determine the phosphorylations of p38 MAPK in the pancreas homogenates. Serum levels of interleukin-1b (IL-1b) and tumor necrosis factor-a (TNF-a) were measured by ELISA; Pathological changes of pancreas were examined and scored with light microscopy.
, 百拇医药
RESULTS: In the SO group, basal p38 MAPK phosphorylation was detected. In the SAP-NS group, the p38 MAPK phosphorylation in the pancreas homogenates reached the maximum at 15 min, remained at a similar level at 30, 60 and 180 min, and declined to the same level as that in SO group at 6 h. In the SAP-NS group and SAP-CNI1493 group, the densities of the band detected by Western blot were 5 200±360, 3 500±250 at 15 min, and 4 910±320, 2 500340 at 30 min (P <0.01). In SAP-CNI1493 group, the serum levels of IL-1b and TNF-a were decreased significantly as compared with those in SAP-NS group (P <0.01). The severity of tissue damage in the SAP-CNI1493 group at 3 h were significantly attenuated in comparison with that of the SAP-NS group (P<0.01).
, 百拇医药
CONCLUSION: The p38 MAPK plays an important role in the pathogenesis of SAP. Inhibition of p38 MAPK phosphorylation may be a potential approach for prevention and treatment of SAP.
Key Words: p38 MAPK; Severe acute pancreatitis; Phosphorylations
Shi XG, Li ZS, Jia YT, Xu YC, Man XH, Gong YF, Tu ZX, Xu GM. p38 mitogen-activated protein kinase signal transduction pathway in pathogenesis of severe acute pancreatitis in rats. Shijie Huaren Xiaohua Zazhi 2005;13(5):653-656
, 百拇医药
摘要
目的: 研究p38丝裂原活化蛋白激酶(p38 MAPK)信号转导通路在大鼠重症急性胰腺炎(SAP)胰腺组织中的变化规律，探讨p38 MAPK特异性抑制剂CNI1493对SAP的保护作用.
方法: 50 g/L牛磺胆酸钠胰胆管逆行注射建立雄性SD大鼠SAP模型，随机分为SO组(假手术组，n = 30)、SAP-NS组(n = 25)及SAP-CNI1493组(n = 25)，各组大鼠质量相似.Western blot法检测大鼠胰腺组织磷酸化p38 MAPK的表达;ELISA方法检测血清IL-1b,TNF-a水平;光镜下评估胰腺组织病理学积分.
结果: SO组胰腺组织存在磷酸化p38 MAPK弱表达，SAP-NS组造模后15 min时磷酸化p38 MAPK的表达即显著增高至峰值，3 h后开始下降，6 h时磷酸化p38 MAPK活性与SO组相似.SAP-NS组和SAP-CNI1493组Western blot检测15，30 min条带光密度值5 200±360, 3 500±250和4 910±320, 2 500±340，SAP-CNI1493组15，30 min时胰腺组织磷酸化p38 MAPK的表达显著低于SAP-NS组(P<0.01).SAP-CNI1493组3，6 h时间点血清IL-1b，TNF-a水平及3 h时间点胰腺组织病理学积分均显著低于SAP-NS组(P<0.01).
, 百拇医药
结论: p38 MAPK信号转导通路与牛磺胆酸钠大鼠SAP发病机制有关，CNI1493预处理可能通过抑制p38 MAPK的激活、减少炎症细胞因子的产生而改善胰腺炎病理改变的严重程度.
关键词: p38丝裂原活化蛋白激酶;重症急性胰腺炎;磷酸化
施新岗, 李兆申, 贾一韬, 许永春, 满晓华, 龚燕芳, 屠振兴, 许国铭. 大鼠重症急性胰腺炎发病机制中p38丝裂原活化蛋白激酶的作用. 世界华人消化杂志 2005;13(5):653-656
参考文献
1 Simeone DM, Zhang L, Graziano K, Nicke B, Pham T, Schaefer C, Logsdon CD. Smad4 mediates activation
, http://www.100md.com
of mitogen-activated protein kinases by TGF- beta in pancreatic acinar cells. Am J Physiol Cell Physiol
2001;281:C311-C319
2 Fleischer F, Dabew R, Goke B, Wagner AC. Stress kinase inhibition modulates acute experimental pancreatitis.
World J Gastroenterol 2002;8:259-265
3 Ren HB, Li ZS, Xu GM, Tu ZX, Shi XG, Jia YT, Gong YF. Dynamic changes of mitogen-activated protein kinase signal
, 百拇医药
transduction in rats with severe acute pancreatitis. Chin J Dig Dis 2004;5:123-125
4 Kyriakis JM, Avruch J . Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress
and inflammation. Physiol Rev 2001;81:807-869
5 Arbabi S, Rosengart MR, Garcia I, Jelacic S, Maier RV. Priming interleukin 8 production: role of platelet-activating
, http://www.100md.com
factor and p38. Arch Surg 1999;134:1348-1353
6 Clifton AD, Young PR, Cohen P. A comparison of the substrate specificity of MAPKAP kinase-2 and MAPKAPkinase-3
and their activation by cytokines and cellular stress. FEBS Lett 1996;392:209-214
7 Janknecht R, Hunter T. Convergence of MAP kinase pathways on the ternary complex factor Sap-1a.
EMBO J 1997;16:1620-1627
, 百拇医药
8 Beyaert R, Cuenda A, Vanden Berghe W, Plaisance S, Lee JC, Haegeman G, Cohen P, Fiers W. The p38/RK
mitogen-activated protein kinase pathway regulates interleukin-6 synthesis response to tumor necrosis factor.
EMBO J 1996;15:1914-1923
9 Dabrowski A, Boguslowicz C, Dabrowska M, Tribillo I, Gabryelewicz A. Reactive oxygen species activate
mitogen-activated protein kinases in pancreatic acinar cells. Pancreas 2000;21:376-384
, 百拇医药
10 O'Neill S, O'Neill AJ, Conroy E, Brady HR, Fitzpatrick JM, Watson RW. Altered caspase expression results in delayed
neutrophil apoptosis in acute pancreatitis. J Leukoc Biol 2000;68:15-20
11 陈浩, 李非, 程韵枫, 孙家邦. 大鼠重症急性胰腺炎病情演变中中性白细胞的作用.
世界华人消化杂志 2001;9:776-779
12 Yan SR, Al-Hertani W, Byers D, Bortolussi R. Lipopolysaccharide-binding protein-and CD14-dependent activation
, 百拇医药
of mitogen-activated protein kinase p38 by lipopolysaccharide in human neutrophils is associated with priming of
respiratory burst. Infect Immun 2002;70:4068-4074
13 English JM, Cobb MH. Pharmacological inhibitors of MAPK pathways. Trends Pharmacol Sci 2002;23:40-45
14 Blinman TA, Gukovsky I, Mouria M, Zaninovic V, Livingston E, Pandol SJ, Gukovskaya AS. Activation of pancreatic
, 百拇医药
acinar cells on isolation from tissue: cytokine upregulation via p38 MAP kinase. Am J Physiol Cell Physiol
2000;279:C1993-2003
15 Ge B, Gram H, Di Padova F, Huang B, New L, Ulevitch RJ, Luo Y, Han J. MAPKK-independent activation of p38alpha
mediated by TAB1-dependent autophosphorylation of p38alpha. Science 2002;295:1291-1294
编辑 潘伯荣 审读 张海宁, 百拇医药( 施新岗,李兆申, 贾一韬,许永春,满晓华, 龚燕芳,屠振兴,许国铭)
施新岗,男, 1970-02-22生, 江苏省启东市人, 汉族, 2004年第二军医大学博士, 主治医师, 主要从事重症急性胰腺炎发病机制和治疗研究
项目负责人: 李兆申，200433，上海市长海路174号，中国人民解放军第二军医大学长海医院消化内科. zhsli@81890.net
电话: 021-25070585
收稿日期: 2004-08-17 接受日期: 2004-10-11
, http://www.100md.com
p38 mitogen-activated protein kinase signal transduction pathway in pathogenesis of severe acute pancreatitis in rats
Xin-Gang Shi, Zhao-Shen Li, Yi-Tao Jia, Yong-Chun Xu, Xiao-Hua Man, Yan-Fang Gong, Zhen-Xing Tu, Guo-Ming Xu
Xin-Gang Shi, Zhao-Shen Li, Yi-Tao Jia, Yong-Chun Xu, Xiao-Hua Man, Yan-Fang Gong, Zhen-Xing Tu, Guo-Ming Xu, Department of Gastroenterology of Changhai Hospital, Second Military Medical University, Shanghai 200433, China
, 百拇医药
Correspondence to: Zhao-Shen Li, Department of Gastroenterology, Changhai Hospital, Second Military Medical University, 174 Changhai Lu, Shanghai 200433, China. zhsli@81890.net
Received: 2004-08-17 Accepted: 2004-10-11
Abstract
AIM: To study the dynamic changes of p38 mitogen-activated protein kinase (p38 MAPK) phosphorylation and to assess the effect of p38 MAPK phosphorylation inhibitor in severe acute pancreatitis (SAP) in rats.
, 百拇医药
METHODS: The SAP model was induced by bili-pancreatic duct retrograde infusion with 5% sterile sodium taurocholate solution. Eighty Sprague-Dawley (SD) rats were randomly divided into sham-operation (SO) group (n = 30), SAP-NS group (n = 25) and SAP-CNI1493 group (n = 25). In the SAP-CNI1493 group, SD rats were administered with 10 mg/kg inhibitor CNI-1493 (i.v.) 30 min before induction of SAP. In SAP-NS group, rats received same volume isotonic saline (i.v.) as CNI-1493. Pancreatic tissues and serum samples were collected before and 15 min, 0.5 h, 1 h, 3 h, 6 h after operation. Western blot analysis was performed to determine the phosphorylations of p38 MAPK in the pancreas homogenates. Serum levels of interleukin-1b (IL-1b) and tumor necrosis factor-a (TNF-a) were measured by ELISA; Pathological changes of pancreas were examined and scored with light microscopy.
, 百拇医药
RESULTS: In the SO group, basal p38 MAPK phosphorylation was detected. In the SAP-NS group, the p38 MAPK phosphorylation in the pancreas homogenates reached the maximum at 15 min, remained at a similar level at 30, 60 and 180 min, and declined to the same level as that in SO group at 6 h. In the SAP-NS group and SAP-CNI1493 group, the densities of the band detected by Western blot were 5 200±360, 3 500±250 at 15 min, and 4 910±320, 2 500340 at 30 min (P <0.01). In SAP-CNI1493 group, the serum levels of IL-1b and TNF-a were decreased significantly as compared with those in SAP-NS group (P <0.01). The severity of tissue damage in the SAP-CNI1493 group at 3 h were significantly attenuated in comparison with that of the SAP-NS group (P<0.01).
, 百拇医药
CONCLUSION: The p38 MAPK plays an important role in the pathogenesis of SAP. Inhibition of p38 MAPK phosphorylation may be a potential approach for prevention and treatment of SAP.
Key Words: p38 MAPK; Severe acute pancreatitis; Phosphorylations
Shi XG, Li ZS, Jia YT, Xu YC, Man XH, Gong YF, Tu ZX, Xu GM. p38 mitogen-activated protein kinase signal transduction pathway in pathogenesis of severe acute pancreatitis in rats. Shijie Huaren Xiaohua Zazhi 2005;13(5):653-656
, 百拇医药
摘要
目的: 研究p38丝裂原活化蛋白激酶(p38 MAPK)信号转导通路在大鼠重症急性胰腺炎(SAP)胰腺组织中的变化规律，探讨p38 MAPK特异性抑制剂CNI1493对SAP的保护作用.
方法: 50 g/L牛磺胆酸钠胰胆管逆行注射建立雄性SD大鼠SAP模型，随机分为SO组(假手术组，n = 30)、SAP-NS组(n = 25)及SAP-CNI1493组(n = 25)，各组大鼠质量相似.Western blot法检测大鼠胰腺组织磷酸化p38 MAPK的表达;ELISA方法检测血清IL-1b,TNF-a水平;光镜下评估胰腺组织病理学积分.
结果: SO组胰腺组织存在磷酸化p38 MAPK弱表达，SAP-NS组造模后15 min时磷酸化p38 MAPK的表达即显著增高至峰值，3 h后开始下降，6 h时磷酸化p38 MAPK活性与SO组相似.SAP-NS组和SAP-CNI1493组Western blot检测15，30 min条带光密度值5 200±360, 3 500±250和4 910±320, 2 500±340，SAP-CNI1493组15，30 min时胰腺组织磷酸化p38 MAPK的表达显著低于SAP-NS组(P<0.01).SAP-CNI1493组3，6 h时间点血清IL-1b，TNF-a水平及3 h时间点胰腺组织病理学积分均显著低于SAP-NS组(P<0.01).
, 百拇医药
结论: p38 MAPK信号转导通路与牛磺胆酸钠大鼠SAP发病机制有关，CNI1493预处理可能通过抑制p38 MAPK的激活、减少炎症细胞因子的产生而改善胰腺炎病理改变的严重程度.
关键词: p38丝裂原活化蛋白激酶;重症急性胰腺炎;磷酸化
施新岗, 李兆申, 贾一韬, 许永春, 满晓华, 龚燕芳, 屠振兴, 许国铭. 大鼠重症急性胰腺炎发病机制中p38丝裂原活化蛋白激酶的作用. 世界华人消化杂志 2005;13(5):653-656
参考文献
1 Simeone DM, Zhang L, Graziano K, Nicke B, Pham T, Schaefer C, Logsdon CD. Smad4 mediates activation
, http://www.100md.com
of mitogen-activated protein kinases by TGF- beta in pancreatic acinar cells. Am J Physiol Cell Physiol
2001;281:C311-C319
2 Fleischer F, Dabew R, Goke B, Wagner AC. Stress kinase inhibition modulates acute experimental pancreatitis.
World J Gastroenterol 2002;8:259-265
3 Ren HB, Li ZS, Xu GM, Tu ZX, Shi XG, Jia YT, Gong YF. Dynamic changes of mitogen-activated protein kinase signal
, 百拇医药
transduction in rats with severe acute pancreatitis. Chin J Dig Dis 2004;5:123-125
4 Kyriakis JM, Avruch J . Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress
and inflammation. Physiol Rev 2001;81:807-869
5 Arbabi S, Rosengart MR, Garcia I, Jelacic S, Maier RV. Priming interleukin 8 production: role of platelet-activating
, http://www.100md.com
factor and p38. Arch Surg 1999;134:1348-1353
6 Clifton AD, Young PR, Cohen P. A comparison of the substrate specificity of MAPKAP kinase-2 and MAPKAPkinase-3
and their activation by cytokines and cellular stress. FEBS Lett 1996;392:209-214
7 Janknecht R, Hunter T. Convergence of MAP kinase pathways on the ternary complex factor Sap-1a.
EMBO J 1997;16:1620-1627
, 百拇医药
8 Beyaert R, Cuenda A, Vanden Berghe W, Plaisance S, Lee JC, Haegeman G, Cohen P, Fiers W. The p38/RK
mitogen-activated protein kinase pathway regulates interleukin-6 synthesis response to tumor necrosis factor.
EMBO J 1996;15:1914-1923
9 Dabrowski A, Boguslowicz C, Dabrowska M, Tribillo I, Gabryelewicz A. Reactive oxygen species activate
mitogen-activated protein kinases in pancreatic acinar cells. Pancreas 2000;21:376-384
, 百拇医药
10 O'Neill S, O'Neill AJ, Conroy E, Brady HR, Fitzpatrick JM, Watson RW. Altered caspase expression results in delayed
neutrophil apoptosis in acute pancreatitis. J Leukoc Biol 2000;68:15-20
11 陈浩, 李非, 程韵枫, 孙家邦. 大鼠重症急性胰腺炎病情演变中中性白细胞的作用.
世界华人消化杂志 2001;9:776-779
12 Yan SR, Al-Hertani W, Byers D, Bortolussi R. Lipopolysaccharide-binding protein-and CD14-dependent activation
, 百拇医药
of mitogen-activated protein kinase p38 by lipopolysaccharide in human neutrophils is associated with priming of
respiratory burst. Infect Immun 2002;70:4068-4074
13 English JM, Cobb MH. Pharmacological inhibitors of MAPK pathways. Trends Pharmacol Sci 2002;23:40-45
14 Blinman TA, Gukovsky I, Mouria M, Zaninovic V, Livingston E, Pandol SJ, Gukovskaya AS. Activation of pancreatic
, 百拇医药
acinar cells on isolation from tissue: cytokine upregulation via p38 MAP kinase. Am J Physiol Cell Physiol
2000;279:C1993-2003
15 Ge B, Gram H, Di Padova F, Huang B, New L, Ulevitch RJ, Luo Y, Han J. MAPKK-independent activation of p38alpha
mediated by TAB1-dependent autophosphorylation of p38alpha. Science 2002;295:1291-1294
编辑 潘伯荣 审读 张海宁, 百拇医药( 施新岗,李兆申, 贾一韬,许永春,满晓华, 龚燕芳,屠振兴,许国铭)
