CO_2气腹对重症急性胰腺炎大鼠炎症因子NF-κB、IL-1、IL-6、TNF-α的影响
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摘要
目的:通过研究CO_2气腹时重症急性胰腺炎(Severe acute pancretitis,SAP)模型大鼠胰腺病变组织及炎症因子水平的改变情况,试探讨CO_2气腹对SAP大鼠炎症反应的影响,为腹腔镜在SAP治疗中的应用提供理论依据。
方法:成年雄性SD大鼠50只,随机分为三组:(1)CO_2气腹组,20只,5%牛磺胆酸钠胆胰管逆行注射方法制备重症急性胰腺炎动物模型,以腹腔镜气腹机向大鼠腹腔内注入CO_2气体,压力12mmHg,维持30分钟;(2)开腹组,20只,建模后关腹,不充入CO_2气体;(3)对照组,10只,仅开腹翻动胰腺后关腹。于术后两个半小时处死各组实验动物。开腹切取胰腺组织行病理学普通光镜及电镜检查,免疫组化检测胰腺组织NF-кB,并取下腔静脉血,放射免疫法检测白介素-1(IL-1)、白介素-6(IL-6)及肿瘤坏死因子-α(TNF-α),仪器法检测血清淀粉酶,比较并分析三组实验数据。
结果:
(1)CO_2气腹组和开腹组胰腺病理学普通光镜及电镜均示灶性胰腺细胞坏死、炎性细胞侵润及出血,两组间病理学评分并无统计学意义(P>0.05);
(2)CO_2气腹组NF-кB、TNF-α及血清淀粉酶与开腹组比较无明显差别(P>0.05),但CO_2气腹组IL-1、IL-6较开腹组明显降低,且具有统计学意义(P<0.05)。
结论:
(1)CO_2气腹对SAP大鼠的胰腺病理改变、血NF-кB、TNF-α水平及血清淀粉酶水平无明显影响。
(2)SAP大鼠血IL-1和IL-6的水平在CO_2气腹影响下有所降低。
Objective: This study used an animal model of Severe acute pancreatitis(SAP) to examine the effects of CO_2 pneumoperitoneum on SAP in rats.And try to explore CO_2 pneumoperitoneum on SAP inflammatory response, as in laparoscopic treatment of SAP applications based on the theory.
Methods: 50 healthy male SD rats were randomly divided into 3 groups:(1)CO_2 pneumoperitoneum group,which include 20 rats. SAP was induced by injection of 5% sodium taurocholate through retrogradely common biliopancreatic ducts via duodenal papilla with epidural catheter,closed puncture hole with medical sealant glue. Then gave CO_2 pneumoperitoneum at a pressure of 12 mmHg for 30 minutes. (2) Open abdominal cavity group,which include 20 rats.The animals were treated as same as CO_2 pneumoperitoneum group except CO_2 pneumoperitoneu. (3) Control group, sham operation, open abdomen for 30 minutes, and then the abdomen was closed. Two and a half hours after the surgical procedures, all animals were killed and cut pancreatic tissue histopathologic examination pathology optical microscope and electron microscope, immunohistochemical detection of pancreatic NF-кB. The blood samples were gathered for examining serum IL-1、IL-6、TNF-α、amylase. Comparison and analysis of three sets of experimental data.
Results: (1) Ordinary light microscope and electron microscopy about pancreatic pathology of CO_2 pneumoperitoneum group and open abdominal cavity group show that there are focal pancreatic necrosis, inflammatory cell invasive and hemorrhage, and have high pathology scores. There were significant difference between the two groups with the control group (P<0.05). there was no statistically important differences between CO_2 pneumoperitoneum group and the open group (P>0.05).(2) NF-кB, IL-1, IL-6, TNF-α, serum amylase were significantly higher in CO_2 pneumoperitoneum group and open abdominal cavity group compared to the control group (P <0.05). There were no statistically important differences between the CO_2 pneumoperitoneum group and open group for NF-кB, TNF-α, serum amylase (P>0.05). However IL-1, IL-6 was significantly lower in CO_2 pneumoperitoneum group compared to the open abdominal cavity group (P<0.05) .
Conclusions:
(1) CO_2 pneumoperitoneum had no significant effect on the inflammatory response in severe acute pancreatitis rats, such as pathology of the pancreas, NF-кB, TNF-αand serum amylase.
(2) CO_2 pneumoperitoneum can be significantly reduced the level of IL-1 and IL-6 expression in the rats with severe acute pancreatitis.
方法:成年雄性SD大鼠50只,随机分为三组:(1)CO_2气腹组,20只,5%牛磺胆酸钠胆胰管逆行注射方法制备重症急性胰腺炎动物模型,以腹腔镜气腹机向大鼠腹腔内注入CO_2气体,压力12mmHg,维持30分钟;(2)开腹组,20只,建模后关腹,不充入CO_2气体;(3)对照组,10只,仅开腹翻动胰腺后关腹。于术后两个半小时处死各组实验动物。开腹切取胰腺组织行病理学普通光镜及电镜检查,免疫组化检测胰腺组织NF-кB,并取下腔静脉血,放射免疫法检测白介素-1(IL-1)、白介素-6(IL-6)及肿瘤坏死因子-α(TNF-α),仪器法检测血清淀粉酶,比较并分析三组实验数据。
结果:
(1)CO_2气腹组和开腹组胰腺病理学普通光镜及电镜均示灶性胰腺细胞坏死、炎性细胞侵润及出血,两组间病理学评分并无统计学意义(P>0.05);
(2)CO_2气腹组NF-кB、TNF-α及血清淀粉酶与开腹组比较无明显差别(P>0.05),但CO_2气腹组IL-1、IL-6较开腹组明显降低,且具有统计学意义(P<0.05)。
结论:
(1)CO_2气腹对SAP大鼠的胰腺病理改变、血NF-кB、TNF-α水平及血清淀粉酶水平无明显影响。
(2)SAP大鼠血IL-1和IL-6的水平在CO_2气腹影响下有所降低。
Objective: This study used an animal model of Severe acute pancreatitis(SAP) to examine the effects of CO_2 pneumoperitoneum on SAP in rats.And try to explore CO_2 pneumoperitoneum on SAP inflammatory response, as in laparoscopic treatment of SAP applications based on the theory.
Methods: 50 healthy male SD rats were randomly divided into 3 groups:(1)CO_2 pneumoperitoneum group,which include 20 rats. SAP was induced by injection of 5% sodium taurocholate through retrogradely common biliopancreatic ducts via duodenal papilla with epidural catheter,closed puncture hole with medical sealant glue. Then gave CO_2 pneumoperitoneum at a pressure of 12 mmHg for 30 minutes. (2) Open abdominal cavity group,which include 20 rats.The animals were treated as same as CO_2 pneumoperitoneum group except CO_2 pneumoperitoneu. (3) Control group, sham operation, open abdomen for 30 minutes, and then the abdomen was closed. Two and a half hours after the surgical procedures, all animals were killed and cut pancreatic tissue histopathologic examination pathology optical microscope and electron microscope, immunohistochemical detection of pancreatic NF-кB. The blood samples were gathered for examining serum IL-1、IL-6、TNF-α、amylase. Comparison and analysis of three sets of experimental data.
Results: (1) Ordinary light microscope and electron microscopy about pancreatic pathology of CO_2 pneumoperitoneum group and open abdominal cavity group show that there are focal pancreatic necrosis, inflammatory cell invasive and hemorrhage, and have high pathology scores. There were significant difference between the two groups with the control group (P<0.05). there was no statistically important differences between CO_2 pneumoperitoneum group and the open group (P>0.05).(2) NF-кB, IL-1, IL-6, TNF-α, serum amylase were significantly higher in CO_2 pneumoperitoneum group and open abdominal cavity group compared to the control group (P <0.05). There were no statistically important differences between the CO_2 pneumoperitoneum group and open group for NF-кB, TNF-α, serum amylase (P>0.05). However IL-1, IL-6 was significantly lower in CO_2 pneumoperitoneum group compared to the open abdominal cavity group (P<0.05) .
Conclusions:
(1) CO_2 pneumoperitoneum had no significant effect on the inflammatory response in severe acute pancreatitis rats, such as pathology of the pancreas, NF-кB, TNF-αand serum amylase.
(2) CO_2 pneumoperitoneum can be significantly reduced the level of IL-1 and IL-6 expression in the rats with severe acute pancreatitis.
引文
[1]李全福,马会敏.腹腔镜技术与腹部手术史、肥胖、妊娠[J].腹腔镜外科杂志,2004,9(2):118-120.
[2] Gaoy G. Applied laparoscopies[J].Antatomy:Abdomen and Pelvis,2004, (12):129 .
[3] Uh W, Warshaw A, Imrie C,et al. IAP guidelines for the surgical management of acute pancreatitis. Pancreatology, 2002,2(6):565-573.
[4]中华医学会外科分会胰腺外科学组.重症急性胰腺炎诊治原则草案[J].中华外科杂志,2001,39(12):963-964.
[5] Norman J. The role of cytokines in the pathogenesis of acute pancreatitis[J]. Am Surg, 1998,175(1):76-83.
[6] KjosseV KT, losanof JE. Laparoscopie treatment of severe acute pancreatitis[J]. Surg Endosc,2001,15(2):272-274.
[7]蔡逊,张兆林,汪训实等.腹腔镜置管行腹腔灌洗治疗重症胰腺炎体会[J].中国现代手术学杂志,2000,4:24-25.
[8] Ona A, Ocampo C, Zandalazini H, et a1. Discussion 141 related articles,books, linkout.Internal drainage of grant acute pseudocysts: the role of video-assisted pancreatic necrosectomy[J].Arch Surg,2000,135:136-140.
[9]嵇武,李令堂,李维勤等.腹腔镜下置管灌洗引流在重症急性胰腺炎早期治疗中的应用[J].中华肝胆外科杂志,2004,12:805-807.
[10] Sargen K, Kingsnorth AN. Management of gallstone pancreatitis: Effects of deviation from clinical guidelines.JOP,2001,2(5):317-322.
[11] Pamoukian VN, Gagner M. Laparoscopic necrosectomy for acute necrotizing pancreatitis[J]. Hepatobiliary Pancreat Surg,2001,8(3):221-223.
[12] Pavars M, IrmejsA, Maurins U, et al. Severe acute pancreatitis: role for laparos- copic surgery[J].Zentralbl Chir,2003,128(10):858-861.
[13]敖翔,陆诚,段宏伟.腹腔镜胆囊切除术二氧化碳气腹对动脉血气和呼气末二氧化碳分压的影响[J].中国临床医学,2004,11(4):637-638.
[14]黄克俭,花人放,孔今城等.静脉输注脂肪乳剂对急性坏死性胰腺炎大鼠脂质炎性介质的影响[J].上海医学,1998,21(12): 700-702.
[15] Yol S, Bostanci EB, Ozogul Y,et al. Effect of carbon dioxide pneumoperitoneum on the severity of acute pancreatitis: an experimental study in rats[J]. Surg Endosc.2004;18(12):1747-1751.
[16] Schmidt J, Lewandrowski K, Fernandez-del Castillo C, et al. Histopathologic correlates of serum amylase activity in acute experimental pancreatitis[J]. Dig D- is Sci,1992,37(9):1426.
[17]朱化刚,耿小平.重症急性胰腺炎胰腺坏死及其外科治疗[J].肝胆外科杂志,2007,15(1):71-72.
[18]何立伟,喻航,张景波,等.重症急性胰腺炎外科干预的时机、指征与方式选择[J].中国现代医学杂志,2004,14(22),114-116
[19] Osman MO,Gesser B,Mortensen JT,et al.Profiles of proinflammatory cytokines in the serum of rabbits after experimentally induced acute pancreatitis[J]. Cytokine,2002.17(1):53.
[20] Kusske AM, Rongione AJ, Reder HA. Cytokines and acute pancreatitis[J]. Gastroenterology,1996;110:639.
[21] T. Hehlgans and K. Pfeffer, The intriguing biology of the tumor necrosis factor/tumor necrosis factor receptor superfamily: players, rules and the games[J].Immunology, 2005,115:1-20.
[22] Fink G, Yang J, Carter G, et al. 1 Acute pancreatitis– induced enzyme release and necrosis are attenuated by IL-1 antagonism through an indirect mechanism [J]. Surg Res,1997,67(1):941.
[23] Jiang CF, Shiau YC, Ngk W, et al. Serum interleukin-6, tumor necrosis factor alpha and C-reactive protein in early prediction of severity of acute pancreatitis [J]. ChinMed Assoc,2004,67:4421.
[24] Yol S, Bostanci EB, Ozogul Y,et al. Effect of carbon dioxide pneumoperitoneum on the severity of acute pancreatitis: an experimental study in rats[J]. Surg Endosc. 2004;18(12):1747-1751.
[25] Jesch NK, Kuebler JF, Nguyen H, et al., Laparoscopy vs minilaparotomy and full laparotomy preserves circulatory but not peritoneal and pulmonary immune responses[J]. Pediatr Surg, 2006,41:1085–1092.
[26] Ure BM, Niewold TA, Bax NMA, et al. Peritoneal, systemic, anddistant organ inflammatory responses are reduced by a laparoscopicapproach and carbon dioxide versus air[J]. Surg Endosc 2002;16:836 - 842.
[27] Champault G, Cruaud P, Guilon P, et al. Is carbon dioxide responsible for the reduction in postoperative infections following laparoscopic surgery[J]. Eur Coelio Surg,1997,3:31.
[28] Kopernik G, Avinoach E, Grossman Y, et al. The effect of a high partial pressure of carbon dioxide environment on metabolism and immune functions of human peritoneal cells—relevance to carbon dioxide pneumoperitoneum[J]. Am Obstet Gynecol 1998;179:1503-1510.
[29] Watson RW, Redond HP, McCarthy J, et al. Exposure of the peritoneal cavity to air regulates early inflammatory responses to surgery in a murine model[J].Br Surg,1995,82:1060-1065.
[30] West MA, Baker J, Bellingham J. Kinetics of decreased LPS-stimulate cytokine release by macrophages exposed to CO2[J]. Surg Res,1996,63:269-274.
[31] Volz J, Koster S, Spacek Z, et al. Characteristic alterations of the peritoneum after carbon dioxide pneumoperitoneum[J]. Surg Endosc,1999,13(6):611.
[1]孙备,姜洪池,许军,等.重症急性胰腺炎手术治疗的经验[J].中华肝胆外科杂志,2001, 7:53-539.
[2] Mithofer K, Muij, ER P R,Warshaw AL. Interventional and surgical treatment of pancreatic abscess[J].Word Surg,1997,21:162-168.
[3] KjosseV KT, losanof JE. Laparoscopie treatment of sevgFg acute pancreatitis[J].Surg Endosc, 2001,15(2):272-274.
[4]孟凡杰,郑德胜,朱冬丽,等.腹腔镜置管灌洗引流治疗急性出血坏死性胰腺炎[J].中华消化内镜杂志,2001,18(5):320.
[5] Chiang DT, Anozie A, Fleming WR, el a1. Comparative study on acute pancreatitis management[J].ANZ Surg,2004,74:218-221.
[6]许力.重症急胜胰腺炎64例治疗体会[J].皖南医学院学报,2006; 25:274-276.
[7] Bagnenko SF, TolstoiAD, Rukhliada NV, et al. Minimally invasive technologies in the treatment of severe forms of acute pancreatitis at various periods of the disease[J]. Vesm Khir Im IIGrek,2002,161(6):30234.
[8]雷若庆,张圣道.全身炎症反应在急性胰腺炎发病机理中的意义[J].中华肝胆外科杂志,2000,6(1):76-77.
[9] Rettally CA, Skarda S, Garza MA, et al. The usefulness of laboratory tests in the early assessment of severity of acute pancreatitis[J]. Crit Rev Clin Lab Sci,2003,40:117-149.
[10] Bismar HA, Al Salamah SM. Outcome of laparoscop cholecystectomy in acute biliary pancreatitis[J]. SaudiMed,2003,24:660-664.
[11] DunkinBJ, PonskyJL, HaleJC. Ultrasound-directed percutaneous endoscopic cystgastrostomy for the treatment of a pancreatic pseudocyst[J]. Surg Endosc,1998,12:1426-1429.
[12] Smadja C, Badawy A, Vons C, et al. Laparoscopic cystogastrostomy for pancreatic pseudo- cyst is safe and effective[J]. Laparoendosc Adv Surg Tech A,1999,9: 401-403.
[13] Seifert H, Dietrich C, Schmitt T, et al. Endoscopic ultrasond guided one step transmural drainage of cystic abdominal lesions with a large channel2 echo endoscope[J]. Endoscopy, 2000,32: 255-259.
[14]张圣道,韩天权,汤耀卿.重症急性胰腺炎临床新技术[M].第1版.北京:人民卫生出版社,2002.
[15]简志祥,陈伟,刘宇斌,等.急性胆源性胰腺炎治疗方式及时机的探讨[J].广东医学, 2002,23 (7):712-713.
[16] Ricci F, Castaldini G, Manzoni G, et a1. Treatment of gall-stone pancreatitis six year experience in a single center[J]. World J Surg,2002,26(12):85.
[17] Kaw M, Antalby AY, Kaw P. Management of gallstone pancreatitis cholecystectomy or ERCP and endoscopic sphinctemtomy [J]. Gastrointest Endose,2002,56:61-65.
[18]李兆申,柏愚,王洛伟.急性胆源性胰腺炎内镜治疗进展[J].国外医学:消化系疾病分册,2004,24 (1):3-4.
[2] Gaoy G. Applied laparoscopies[J].Antatomy:Abdomen and Pelvis,2004, (12):129 .
[3] Uh W, Warshaw A, Imrie C,et al. IAP guidelines for the surgical management of acute pancreatitis. Pancreatology, 2002,2(6):565-573.
[4]中华医学会外科分会胰腺外科学组.重症急性胰腺炎诊治原则草案[J].中华外科杂志,2001,39(12):963-964.
[5] Norman J. The role of cytokines in the pathogenesis of acute pancreatitis[J]. Am Surg, 1998,175(1):76-83.
[6] KjosseV KT, losanof JE. Laparoscopie treatment of severe acute pancreatitis[J]. Surg Endosc,2001,15(2):272-274.
[7]蔡逊,张兆林,汪训实等.腹腔镜置管行腹腔灌洗治疗重症胰腺炎体会[J].中国现代手术学杂志,2000,4:24-25.
[8] Ona A, Ocampo C, Zandalazini H, et a1. Discussion 141 related articles,books, linkout.Internal drainage of grant acute pseudocysts: the role of video-assisted pancreatic necrosectomy[J].Arch Surg,2000,135:136-140.
[9]嵇武,李令堂,李维勤等.腹腔镜下置管灌洗引流在重症急性胰腺炎早期治疗中的应用[J].中华肝胆外科杂志,2004,12:805-807.
[10] Sargen K, Kingsnorth AN. Management of gallstone pancreatitis: Effects of deviation from clinical guidelines.JOP,2001,2(5):317-322.
[11] Pamoukian VN, Gagner M. Laparoscopic necrosectomy for acute necrotizing pancreatitis[J]. Hepatobiliary Pancreat Surg,2001,8(3):221-223.
[12] Pavars M, IrmejsA, Maurins U, et al. Severe acute pancreatitis: role for laparos- copic surgery[J].Zentralbl Chir,2003,128(10):858-861.
[13]敖翔,陆诚,段宏伟.腹腔镜胆囊切除术二氧化碳气腹对动脉血气和呼气末二氧化碳分压的影响[J].中国临床医学,2004,11(4):637-638.
[14]黄克俭,花人放,孔今城等.静脉输注脂肪乳剂对急性坏死性胰腺炎大鼠脂质炎性介质的影响[J].上海医学,1998,21(12): 700-702.
[15] Yol S, Bostanci EB, Ozogul Y,et al. Effect of carbon dioxide pneumoperitoneum on the severity of acute pancreatitis: an experimental study in rats[J]. Surg Endosc.2004;18(12):1747-1751.
[16] Schmidt J, Lewandrowski K, Fernandez-del Castillo C, et al. Histopathologic correlates of serum amylase activity in acute experimental pancreatitis[J]. Dig D- is Sci,1992,37(9):1426.
[17]朱化刚,耿小平.重症急性胰腺炎胰腺坏死及其外科治疗[J].肝胆外科杂志,2007,15(1):71-72.
[18]何立伟,喻航,张景波,等.重症急性胰腺炎外科干预的时机、指征与方式选择[J].中国现代医学杂志,2004,14(22),114-116
[19] Osman MO,Gesser B,Mortensen JT,et al.Profiles of proinflammatory cytokines in the serum of rabbits after experimentally induced acute pancreatitis[J]. Cytokine,2002.17(1):53.
[20] Kusske AM, Rongione AJ, Reder HA. Cytokines and acute pancreatitis[J]. Gastroenterology,1996;110:639.
[21] T. Hehlgans and K. Pfeffer, The intriguing biology of the tumor necrosis factor/tumor necrosis factor receptor superfamily: players, rules and the games[J].Immunology, 2005,115:1-20.
[22] Fink G, Yang J, Carter G, et al. 1 Acute pancreatitis– induced enzyme release and necrosis are attenuated by IL-1 antagonism through an indirect mechanism [J]. Surg Res,1997,67(1):941.
[23] Jiang CF, Shiau YC, Ngk W, et al. Serum interleukin-6, tumor necrosis factor alpha and C-reactive protein in early prediction of severity of acute pancreatitis [J]. ChinMed Assoc,2004,67:4421.
[24] Yol S, Bostanci EB, Ozogul Y,et al. Effect of carbon dioxide pneumoperitoneum on the severity of acute pancreatitis: an experimental study in rats[J]. Surg Endosc. 2004;18(12):1747-1751.
[25] Jesch NK, Kuebler JF, Nguyen H, et al., Laparoscopy vs minilaparotomy and full laparotomy preserves circulatory but not peritoneal and pulmonary immune responses[J]. Pediatr Surg, 2006,41:1085–1092.
[26] Ure BM, Niewold TA, Bax NMA, et al. Peritoneal, systemic, anddistant organ inflammatory responses are reduced by a laparoscopicapproach and carbon dioxide versus air[J]. Surg Endosc 2002;16:836 - 842.
[27] Champault G, Cruaud P, Guilon P, et al. Is carbon dioxide responsible for the reduction in postoperative infections following laparoscopic surgery[J]. Eur Coelio Surg,1997,3:31.
[28] Kopernik G, Avinoach E, Grossman Y, et al. The effect of a high partial pressure of carbon dioxide environment on metabolism and immune functions of human peritoneal cells—relevance to carbon dioxide pneumoperitoneum[J]. Am Obstet Gynecol 1998;179:1503-1510.
[29] Watson RW, Redond HP, McCarthy J, et al. Exposure of the peritoneal cavity to air regulates early inflammatory responses to surgery in a murine model[J].Br Surg,1995,82:1060-1065.
[30] West MA, Baker J, Bellingham J. Kinetics of decreased LPS-stimulate cytokine release by macrophages exposed to CO2[J]. Surg Res,1996,63:269-274.
[31] Volz J, Koster S, Spacek Z, et al. Characteristic alterations of the peritoneum after carbon dioxide pneumoperitoneum[J]. Surg Endosc,1999,13(6):611.
[1]孙备,姜洪池,许军,等.重症急性胰腺炎手术治疗的经验[J].中华肝胆外科杂志,2001, 7:53-539.
[2] Mithofer K, Muij, ER P R,Warshaw AL. Interventional and surgical treatment of pancreatic abscess[J].Word Surg,1997,21:162-168.
[3] KjosseV KT, losanof JE. Laparoscopie treatment of sevgFg acute pancreatitis[J].Surg Endosc, 2001,15(2):272-274.
[4]孟凡杰,郑德胜,朱冬丽,等.腹腔镜置管灌洗引流治疗急性出血坏死性胰腺炎[J].中华消化内镜杂志,2001,18(5):320.
[5] Chiang DT, Anozie A, Fleming WR, el a1. Comparative study on acute pancreatitis management[J].ANZ Surg,2004,74:218-221.
[6]许力.重症急胜胰腺炎64例治疗体会[J].皖南医学院学报,2006; 25:274-276.
[7] Bagnenko SF, TolstoiAD, Rukhliada NV, et al. Minimally invasive technologies in the treatment of severe forms of acute pancreatitis at various periods of the disease[J]. Vesm Khir Im IIGrek,2002,161(6):30234.
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