Effects of δ-Opioid Receptor Agonists on Hepatocyte Apoptosis and Expressions of bcl-2 and caspase-3 in Septic Rat_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

FENG Wenming , ZHU Ming , BAO Yin , TAO Yulong , WANG Yao , ZHANG Xiaolan

Institute of Molecular Surgery, The First People’s Hospital of Huzhou City, Zhejiang Province, Huzhou 313000, China;

Corresponding author：

Keywords：

δ-opioid receptor agonists; Sepsis; Hepatocyty apoptosis; Protein expression; bcl-2; caspase-3

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Objective To observe the effects of δ-opioid receptor agonists D-Ala2-D-Leu5-enkephali （DADLE） on hepatocyte apoptosis and expressions of bcl-2 and caspase-3 in septic rat, and to investigate the possible mechanism by which DADLE protects the liver in sepsis.
Methods Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-four SD rats (either male or female) were randomly divided into CLP group (n=18), DADLE group (n=18) and sham operation (SO) group (n=18). The rats were respectively killed at different time (2 h, 4 h and 6 h after operation). Hepatocyte apoptosis was detected by TdT-mediated dUTP Nick End Labeling (TUNEL). The expressions of bcl-2 and caspase-3 protein were detected by immunohistochemistry. And the changes of pathology in hepatic tissue were detected by light microscope.
Results The hepatic pathological lesion of rats in CLP group was obviously serious compared with SO group, while it was obviously improved in DADLE group. The apoptosis index of rat hepatocytes in CLP group significantly increased compared with SO group, and further it was prominent at 4 h （P＜0.01). The apoptosis index of rat hepatocytes at each time of DADLE group was significantly decreased compared with CLP group （P＜0.01）. Expression of caspase-3 protein in liver tissues of CLP group significantly increased compared with SO group （P＜0.01）, while the expression of bcl-2 protein significantly decreased （P＜0.05）. Expression of caspase-3 protein in liver tissues of DADLE group significantly decreased compared with the CLP group （P＜0.01）, while the expression of bcl-2 protein significantly increased （P＜0.05）. There was positive correlation between expression of caspase-3 in liver tissues and apoptosis index of hepatocyte (r=0.83, P＜0.01) and negative correlation between expression of bcl-2 in liver tissues and apoptosis index of hepatocyte (r=-0.65, P＜0.01).
Conclusions The findings indicate that δ-opioid receptor agonists DADLE can obviously improve hepatic pathological changes of septic rats. And its protective mechanism contains down regulation of caspase-3 expression, upregulation of bcl-2 expression and thus the apoptosis of hepatocyte is repressed.

Citation： FENG Wenming,ZHU Ming,BAO Yin,TAO Yulong,WANG Yao,ZHANG Xiaolan. Effects of δ-Opioid Receptor Agonists on Hepatocyte Apoptosis and Expressions of bcl-2 and caspase-3 in Septic Rat. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(7): 550-554. doi: Copy

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1. 蒋力生，陈鹏. 脓毒症与内毒素血症的治疗现状与展望 [Ｊ]. 中国普外基础与临床杂志， 2004；11（6）： 551553.
2. 姚咏明，盛志勇. 脓毒症研究的若干新动态 [Ｊ]. 中国危重病急救医学, 2000; 12(6): 323325.
3. 李斌，李玉民，李汛，等. 环氧合酶2对脓毒症大鼠肝损伤的影响 [Ｊ]. 中国普外基础与临床杂志， 2008； 15（4）： 266270.
4. 高建芝, 张建龙, 王红梅, 等. 脓毒症大鼠FasL、caspase3和NFκB的表达与肝细胞凋亡的关系 [Ｊ]. 新乡医学院学报, 2004; 21(2): 9699.
5. Harvey NL, Kumar S. The role of caspases in apoptosis[Ｊ].Adv Biochem Eng Biotechnol, 1998; 62(199): 107128.
6. Iwata A, Stevenson VM, Minard A, et al. Overexpression of bcl2 provides protection in septic mice by a trans effect[Ｊ]. J Immunol, 2003; 171(6): 31363141.
7. Chaudry IH, Wichterman KA, Baue AE. Effect of sepsis on tissue adenine nucleotide levels[Ｊ]. Surgery, 1979; 85(2): 205211.
8. 陈绪军，肖琦，艾中立，等. 肝细胞损伤，细胞凋亡及肝细胞保护（文献综述） [Ｊ]. 国外医学外科学分册, 2000; 27(1): 2124.
9. 张毅，叶启发，周进学，等. 三七总皂苷对大鼠肝移植物细胞凋亡及bcl2和caspase3表达的影响 [Ｊ]. 中国普外基础与临床杂志， 2005； 12（2）： 113116.
10. 肖旭, 梅广林, 胡卫东, 等. 葛根素对大鼠肝缺血再灌注后caspase3和bcl2表达的影响 [Ｊ]. 交通医学, 2008; 22(5): 472473.
11. Tsamandas AC, Thomopoulos K, Zolota V, et al. Potential role of bcl2 and bax mRNA and protein expression in chronic hepatitis type B and C: a clinicopathologic study[Ｊ]. Mod Pathol, 2003; 16(12): 12731288.
12. Zhang M, He W, Liu F, et al. Inhibition of mouse hepatocyte apoptosis via antiFas ribozyme[Ｊ]. World J Gastroenterol, 2004; 10(17): 25672570.
13. Kienle K, Rentsch M, Müller T, et al. Expression of BCL2 in liver grafts after adenoviral transfer improves survival following prolonged ischemia and reperfusion in rat liver transplantation[Ｊ]. Transplant Proc, 2005; 37(1): 439441.
14. Wang XZ, Zhang SJ, Chen YX, et al. Effects of plateletderived growth factor and interleukin10 on Fas/Fasligand and Bcl2/Bax mRNA expression in rat hepatic stellate cells in vitro[Ｊ]. World J Gastroenterol, 2004; 10(18): 27062710.
15. Harris MH, Thompson CB.The role of the Bcl2 family in the regulation of outer mitochondrial membrane permeability[Ｊ]. Cell Death Differ, 2000; 7(12): 11821191.
16. 董月青, 黄军华, 姚咏明. 脓毒症中T淋巴细胞凋亡及其调控研究进展 [Ｊ]. 中国危重病急救医学, 2004; 16(6): 381384.
17. Yang J, Gallagher SF, Haines K, et al. Kupffer cellderived Fas ligand plays a role in liver injury and hepatocyte death[Ｊ]. J Gastrointest Surg, 2004; 8(2): 166174.
18. 付小兵，杨银辉，孙同柱，等. 缺血再灌注诱导bcl2基因表达及其对肠道细胞凋亡的影响 [Ｊ]. 中国危重病急救医学, 1999; 11(8): 459461.
19. Adams JM, Cory S. The Bcl2 protein family: arbiters of cell survival [Ｊ]. Science, 1998; 281(5381): 13221326.
20. 王玉梅, 冯国和, 刘德刚, 等. Bcl2蛋白及caspase3基因表达与暴发性肝衰竭肝细胞凋亡关系的研究 [Ｊ]. 中国医科大学学报, 2006; 35(5): 494496.
21. 王玉梅, 冯国和, 黄芬, 等. 肿瘤坏死因子α及caspase3表达与暴发性肝衰竭细胞凋亡 [Ｊ]. 中华内科杂志, 2003; 42(8): 566570.
22. Yuste VJ, SánchezLópez I, Solé C, et al. The contribution of apoptosisinducing factor, caspaseactivated DNase, and inhibitor of caspaseactivated DNase to the nuclear phenotype and DNA degradation during apoptosis [Ｊ]. J Biol Chem, 2005; 280(42): 3567035683.
23. Fumarola C, Guidotti GG. Stressinduced apoptosis: toward a symmetry with receptormediated cell death [Ｊ]. Apoptosis, 2004; 9(1): 7782.
24. Meier P, Finch A, Evan G. Apoptosis in development[Ｊ]. Nature, 2000; 407(6805): 796801.
25. Gross GJ. Role of opioids in acute and delayed preconditioning [Ｊ]. J Mol Cell Cardiol, 2003; 35(7): 709718.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effects of δ-Opioid Receptor Agonists on Hepatocyte Apoptosis and Expressions of bcl-2 and caspase-3 in Septic Rat

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