Effect on Expression of High Mobility Group Box-1 mRNA for Expressions of Zonula Occludens Protein-1 mRNA and Protein in Rat with Acute Necrotizing Pancreatitis_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 LUAN Zhenggang ¹ , GUO Renxuan ²

1.Department of Intensive Care Unit， The First Affiliated Hospital of China Medical University， Shenyang 110001， Liaoning Province， China;;
2.Department of General Surgery， The First Affiliated Hospital of China Medical University， Shenyang 110001， Liaoning Province， China;

Corresponding author：

LUAN Zhenggang, Email: zgl17698@sina.com

Keywords：

Acute necrotizing pancreatitis; High mobility group box-1 protein; Zonula occludens protein-1; Intestinal mucosal barrier; Rat

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Objective 　To study the effect on expression of high mobility group box-1 （HMGB1） mRNA for the expression of zonula occludens-1 （ZO-1） in ileum tissues， and to explore the possible mechanism of intestinal mucosal barrier injury in rats with acute necrotizing pancreatitis （ANP）.
Methods 　Ninety-six male Wistar rats were divided randomly （random number method） into ANP group， ethyl pyruvate （EP）group， and sham operation group. Eight rats of 3 groups were killed to get abdominal aortic blood and ileal tissues at 6， 12， 24， and 48h after operation， respectively.The levels of plasma amylase （AMY）， D-lactate acid， and the activity of malonyl dialdehyde （MDA） in the ileum tissues were determined by using automatic biochemical analyzer， improved enzymatic spectrophotometry， and thiobarbituric acid （TAB） colorimetry respectively. The pathological changes of ileum tissues were observed under microscopy by HE staining， the expression of ZO-1 protein in ileum tissues was observed by immunohistochemistry （SP method）， and the expressions of HMGB1 mRNA and ZO-1 mRNA in ileum tissues were detected by reverse transcription-polymerase chain reaction （RT-PCR）.
Results 　Compared with ANP group at the same time， levels of AMY， D-lactate acid， and MDA in ileum tissues of EP group were all significantly lower （P＜0.05）. The expression level of HMGB1 mRNA increased at 6 h while ZO-1 mRNA decreased in ANP group. Compared with ANP group at the same time， the expression level of HMGB1 mRNA of EP group was significantly lower while ZO-1 mRNA was higher （P＜0.05）， and the pathological damage in ileum tissues was lighter.
Conclusions 　The decreased expression of ZO-1 in ileum tissues is one of the vital
causes for intestinal mucosal barrier injury in ANP， and it probably occurs in case of the excessive expression of HMGB1.

Citation： LUAN Zhenggang,GUO Renxuan. Effect on Expression of High Mobility Group Box-1 mRNA for Expressions of Zonula Occludens Protein-1 mRNA and Protein in Rat with Acute Necrotizing Pancreatitis. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2013, 20(3): 265-270. doi: Copy

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10.	Wang H， Yang H， Tracey KJ. Extracellular role of HMGB1 in infla-mmation and sepsis[J]. J Intern Med， 2004， 255(3)：320-331.
11.	Luan ZG， Zhang H， Ma XC， et al. Role of high-mobility group box 1 protein in the pathogenesis of intestinal barrier injury in rats with severe acute pancreatitis[J]. Pancreas， 2010， 39(2)：216-223.
12.	Aho HJ， Koskensalo SM， Nevalainen TJ. Experimental pancreatitisin the rat. Sodium taurocholate-induced acute haemorrhagic pancreatitis[J]. Scand J Gastroenterol， 1980， 15(4)：411-416.
13.	Gerlach H. Risk management in patients with severe acute pancreatitis[J]. Crit Care， 2004， 8(6)：430-432.
14.	Heinrich S， Schäfer M， Rousson V， et al. Evidence-based treatment of acute pancreatitis：a look at established paradigms[J]. Ann Surg， 2006， 243(2)：154-168.
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20.	Nusrat A， Turner JR， Madara JL. Molecular physiology and pathophysiology of tight junctions.Ⅳ. Regulation of tight junctions by extracellular stimuli：nutrients， cytokines， and immune cells[J]. Am J Physiol Gastrointest Liver Physiol， 2000， 279(5)：G851-G857.
21.	秦勇，党晓燕. 白藜芦醇对重症急性胰腺炎大鼠肠黏膜缺血-再灌注损伤的保护作用[J]. 中国普外基础与临床杂志， 2009， 16(10)：819-823.
22.	Mazzon E， Sturniolo GC， Puzzolo D， et al. Effect of stress on the paracellular barrier in the rat ileum[J]. Gut， 2002， 51(4)：507-513.
23.	Stevenson BR， Siliciano JD， Mooseker MS， et al. Identification of ZO-1：a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia[J]. J Cell Biol， 1986， 103(3)：755-766.
24.	Fink MP. Ethyl pyruvate：a novel anti-inflammatory agent[J]. J Intern Med， 2007， 261(4)：349-362.
25.	Yang R， Uchiyama T， Alber SM， et al. Ethyl pyruvate ameliorates distant organ injury in a murine model of acute necrotizing pancreatitis[J]. Crit Care Med， 2004， 32(7)：1453-1459.
26.	Ulloa L， Ochani M， Yang H， et al. Ethyl pyruvate prevents lethalityin mice with established lethal sepsis and systemic inflammation[J]. Proc Natl Acad Sci U S A， 2002， 99(19)：12351-12356.
27.	， 41(7)：1080-1085.

1. Pitchumoni CS， Patel NM， Shah P. Factors influencing mortality in acute pancreatitis：can we alter them?[J]. J Clin Gastroenterol， 2005， 39(9)：798-814.
2. Bank S， Singh P， Pooran N， et al. Evaluation of factors that have reduced mortality from acute pancreatitis over the past 20 years[J]. J Clin Gastroenterol， 2002， 35(1)：50-60.
3. Jamdar S， Siriwardena AK. Contemporary management ofinfected necrosis complicating severe acute pancreatitis[J]. Crit Care， 2006， 10(1)：101.
4. 刁永鹏，陈宏，李非. 重症急性胰腺炎肠屏障功能障碍的研究进展[J]. 中国普外基础与临床杂志， 2009， 16(7)：588-592.
5. 王琛，樊勇. 急性胰腺炎常见并发症及防治[J]. 中国普外基础与临床杂志， 2010， 17(10)：1107-1108.
6. Blikslager AT， Moeser AJ， Gookin JL， et al. Restoration of barrier function in injured intestinal mucosa[J]. Physiol Rev， 2007， 87(2)：545-564.
7. Li Q， Zhang Q， Wang M， et al. Interferon-gamma and tumor necrosis factor-alpha disrupt epithelial barrier function by alteringlipid composition in membrane microdomains of tight junction[J]. Clin Immunol， 2008， 126(1)：67-80.
8. 曹洪庆，黄鹤光，陈燕昌，等. 重症急性胰腺炎大鼠紧密连接蛋白-1的表达与肠黏膜屏障损伤[J]. 中国普外基础与临床杂志， 2011， 18(3)：240-244.
9. 高志光，秦环龙. 肠上皮细胞紧密连接的生物学功能及在肠屏障中的作用[J]. 肠外与肠内营养， 2005， 12(5)：299-302.
10. Wang H， Yang H， Tracey KJ. Extracellular role of HMGB1 in infla-mmation and sepsis[J]. J Intern Med， 2004， 255(3)：320-331.
11. Luan ZG， Zhang H， Ma XC， et al. Role of high-mobility group box 1 protein in the pathogenesis of intestinal barrier injury in rats with severe acute pancreatitis[J]. Pancreas， 2010， 39(2)：216-223.
12. Aho HJ， Koskensalo SM， Nevalainen TJ. Experimental pancreatitisin the rat. Sodium taurocholate-induced acute haemorrhagic pancreatitis[J]. Scand J Gastroenterol， 1980， 15(4)：411-416.
13. Gerlach H. Risk management in patients with severe acute pancreatitis[J]. Crit Care， 2004， 8(6)：430-432.
14. Heinrich S， Schäfer M， Rousson V， et al. Evidence-based treatment of acute pancreatitis：a look at established paradigms[J]. Ann Surg， 2006， 243(2)：154-168.
15. Ammori BJ. Role of the gut in the course of severe acute pancreatitis[J]. Pancreas， 2003， 26(2)：122-129.
16. Dervenis C， Smailis D， Hatzitheoklitos E. Bacterial translocation and its prevention in acute pancreatitis[J]. J Hepatobiliary Pancreat Surg， 2003， 10(6)：415-418.
17. Furuya T， Soeno T， Komatsu M. Strategy for bacterial translocation in acute pancreatitis[J]. Nippon Shokakibyo Gakkai Zasshi， 2004， 101(5)：502-509.
18. Chen X， Zhao HX， Fu XS， et al. Glucagonlike peptide 2 protectsintestinal barrier in severe acute pancreatitis through regulating intestinal epithelial cell proliferation and apoptosis[J]. Pancreas，.
19. Wang X， Wang B， Wu J， et al. Beneficial effects of growth hormone on bacterial translocation during the course of acute necroti-zing pancreatitis in rats[J]. Pancreas， 2001， 23(2)：148-156.
20. Nusrat A， Turner JR， Madara JL. Molecular physiology and pathophysiology of tight junctions.Ⅳ. Regulation of tight junctions by extracellular stimuli：nutrients， cytokines， and immune cells[J]. Am J Physiol Gastrointest Liver Physiol， 2000， 279(5)：G851-G857.
21. 秦勇，党晓燕. 白藜芦醇对重症急性胰腺炎大鼠肠黏膜缺血-再灌注损伤的保护作用[J]. 中国普外基础与临床杂志， 2009， 16(10)：819-823.
22. Mazzon E， Sturniolo GC， Puzzolo D， et al. Effect of stress on the paracellular barrier in the rat ileum[J]. Gut， 2002， 51(4)：507-513.
23. Stevenson BR， Siliciano JD， Mooseker MS， et al. Identification of ZO-1：a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia[J]. J Cell Biol， 1986， 103(3)：755-766.
24. Fink MP. Ethyl pyruvate：a novel anti-inflammatory agent[J]. J Intern Med， 2007， 261(4)：349-362.
25. Yang R， Uchiyama T， Alber SM， et al. Ethyl pyruvate ameliorates distant organ injury in a murine model of acute necrotizing pancreatitis[J]. Crit Care Med， 2004， 32(7)：1453-1459.
26. Ulloa L， Ochani M， Yang H， et al. Ethyl pyruvate prevents lethalityin mice with established lethal sepsis and systemic inflammation[J]. Proc Natl Acad Sci U S A， 2002， 99(19)：12351-12356.
27. ， 41(7)：1080-1085.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect on Expression of High Mobility Group Box-1 mRNA for Expressions of Zonula Occludens Protein-1 mRNA and Protein in Rat with Acute Necrotizing Pancreatitis

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