• 浙江省湖州市第一人民醫(yī)院分子外科研究所(浙江湖州 313000);

目的  觀察δ阿片受體激動(dòng)劑D-丙2,D-亮5腦啡肽(D-Ala2,D-Leu5-enkephali,DADLE)對(duì)膿毒癥大鼠肝細(xì)胞凋亡及肝組織bcl-2和caspase-3表達(dá)的影響,探討DADLE對(duì)肝臟保護(hù)作用的可能機(jī)理。
方法  采用盲腸結(jié)扎加穿孔(CLP)法制作大鼠膿毒癥模型,SD大鼠54只(雌雄不限),采用隨機(jī)數(shù)字表法隨機(jī)分為CLP組(n=18)、DADLE組(n=18)和假手術(shù)組(n=18)。在不同時(shí)間點(diǎn)(2、4及6 h)處死大鼠,原位末端標(biāo)記法檢測(cè)肝細(xì)胞凋亡情況,免疫組化法檢測(cè)肝組織中bcl-2及caspase-3蛋白表達(dá)的動(dòng)態(tài)變化并觀察肝臟的病理改變。
結(jié)果  CLP組大鼠肝組織病理?yè)p害明顯較假手術(shù)組嚴(yán)重,DADLE組肝臟的病理變化明顯改善; CLP組大鼠肝組織細(xì)胞凋亡指數(shù)明顯較假手術(shù)組升高(P<0.01),以4 h最顯著(P<0.01),DADLE 組各時(shí)相肝細(xì)胞凋亡指數(shù)均明顯降低(P<0.01); CLP組大鼠肝組織caspase3蛋白的表達(dá)強(qiáng)度比假手術(shù)組明顯增強(qiáng)(P<0.01),而bcl-2蛋白的表達(dá)則明顯減弱(P<0.05); DADLE組肝組織caspase-3蛋白的表達(dá)強(qiáng)度較CLP組明顯減弱(P<0.01),而bcl-2蛋白的表達(dá)則明顯增強(qiáng)(P<0.05)。肝組織caspase-3的表達(dá)與肝細(xì)胞凋亡指數(shù)呈正相關(guān)(r=0.83,P<0.01),bcl-2的表達(dá)與肝細(xì)胞凋亡指數(shù)呈負(fù)相關(guān)(r=-0.65,P<0.01)。
結(jié)論  DADLE能明顯改善膿毒癥大鼠肝臟的病理變化,其作用機(jī)理可能與DADLE下調(diào)caspase-3表達(dá)、上調(diào)bcl-2表達(dá),從而抑制肝細(xì)胞凋亡有關(guān)。

引用本文: 馮文明,朱鳴,鮑鷹,陶玉龍,王耀,張曉嵐. δ阿片受體激動(dòng)劑對(duì)膿毒癥大鼠肝細(xì)胞凋亡及肝組織bcl-2和caspase-3表達(dá)的影響. 中國(guó)普外基礎(chǔ)與臨床雜志, 2009, 16(7): 550-554. doi: 復(fù)制

1. 蔣力生, 陳鵬. 膿毒癥與內(nèi)毒素血癥的治療現(xiàn)狀與展望 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2004;11(6): 551553.
2. 姚詠明, 盛志勇. 膿毒癥研究的若干新動(dòng)態(tài) [J]. 中國(guó)危重病急救醫(yī)學(xué), 2000; 12(6): 323325.
3. 李斌, 李玉民, 李汛, 等. 環(huán)氧合酶2對(duì)膿毒癥大鼠肝損傷的影響 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008; 15(4): 266270.
4. 高建芝, 張建龍, 王紅梅, 等. 膿毒癥大鼠FasL、caspase3和NFκB的表達(dá)與肝細(xì)胞凋亡的關(guān)系 [J]. 新鄉(xiāng)醫(yī)學(xué)院學(xué)報(bào), 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. 陳緒軍, 肖琦, 艾中立, 等. 肝細(xì)胞損傷, 細(xì)胞凋亡及肝細(xì)胞保護(hù)(文獻(xiàn)綜述) [J]. 國(guó)外醫(yī)學(xué)外科學(xué)分冊(cè), 2000; 27(1): 2124.
9. 張毅, 葉啟發(fā), 周進(jìn)學(xué), 等. 三七總皂苷對(duì)大鼠肝移植物細(xì)胞凋亡及bcl2和caspase3表達(dá)的影響 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2005; 12(2): 113116.
10. 肖旭, 梅廣林, 胡衛(wèi)東, 等. 葛根素對(duì)大鼠肝缺血再灌注后caspase3和bcl2表達(dá)的影響 [J]. 交通醫(yī)學(xué), 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, 黃軍華, 姚詠明. 膿毒癥中T淋巴細(xì)胞凋亡及其調(diào)控研究進(jìn)展 [J]. 中國(guó)危重病急救醫(yī)學(xué), 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, 楊銀輝, 孫同柱, 等. 缺血再灌注誘導(dǎo)bcl2基因表達(dá)及其對(duì)腸道細(xì)胞凋亡的影響 [J]. 中國(guó)危重病急救醫(yī)學(xué), 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, 馮國(guó)和, 劉德剛, 等. Bcl2蛋白及caspase3基因表達(dá)與暴發(fā)性肝衰竭肝細(xì)胞凋亡關(guān)系的研究 [J]. 中國(guó)醫(yī)科大學(xué)學(xué)報(bào), 2006; 35(5): 494496.
21. 王玉梅, 馮國(guó)和, 黃芬, 等. 腫瘤壞死因子α及caspase3表達(dá)與暴發(fā)性肝衰竭細(xì)胞凋亡 [J]. 中華內(nèi)科雜志, 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.
  1. 1. 蔣力生, 陳鵬. 膿毒癥與內(nèi)毒素血癥的治療現(xiàn)狀與展望 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2004;11(6): 551553.
  2. 2. 姚詠明, 盛志勇. 膿毒癥研究的若干新動(dòng)態(tài) [J]. 中國(guó)危重病急救醫(yī)學(xué), 2000; 12(6): 323325.
  3. 3. 李斌, 李玉民, 李汛, 等. 環(huán)氧合酶2對(duì)膿毒癥大鼠肝損傷的影響 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008; 15(4): 266270.
  4. 4. 高建芝, 張建龍, 王紅梅, 等. 膿毒癥大鼠FasL、caspase3和NFκB的表達(dá)與肝細(xì)胞凋亡的關(guān)系 [J]. 新鄉(xiāng)醫(yī)學(xué)院學(xué)報(bào), 2004; 21(2): 9699.
  5. 5. Harvey NL, Kumar S. The role of caspases in apoptosis[J].Adv Biochem Eng Biotechnol, 1998; 62(199): 107128.
  6. 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. 7. Chaudry IH, Wichterman KA, Baue AE. Effect of sepsis on tissue adenine nucleotide levels[J]. Surgery, 1979; 85(2): 205211.
  8. 8. 陳緒軍, 肖琦, 艾中立, 等. 肝細(xì)胞損傷, 細(xì)胞凋亡及肝細(xì)胞保護(hù)(文獻(xiàn)綜述) [J]. 國(guó)外醫(yī)學(xué)外科學(xué)分冊(cè), 2000; 27(1): 2124.
  9. 9. 張毅, 葉啟發(fā), 周進(jìn)學(xué), 等. 三七總皂苷對(duì)大鼠肝移植物細(xì)胞凋亡及bcl2和caspase3表達(dá)的影響 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2005; 12(2): 113116.
  10. 10. 肖旭, 梅廣林, 胡衛(wèi)東, 等. 葛根素對(duì)大鼠肝缺血再灌注后caspase3和bcl2表達(dá)的影響 [J]. 交通醫(yī)學(xué), 2008; 22(5): 472473.
  11. 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. 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. 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. 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. 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. 16. 董月青, 黃軍華, 姚詠明. 膿毒癥中T淋巴細(xì)胞凋亡及其調(diào)控研究進(jìn)展 [J]. 中國(guó)危重病急救醫(yī)學(xué), 2004; 16(6): 381384.
  17. 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. 18. 付小兵, 楊銀輝, 孫同柱, 等. 缺血再灌注誘導(dǎo)bcl2基因表達(dá)及其對(duì)腸道細(xì)胞凋亡的影響 [J]. 中國(guó)危重病急救醫(yī)學(xué), 1999; 11(8): 459461.
  19. 19. Adams JM, Cory S. The Bcl2 protein family: arbiters of cell survival [J]. Science, 1998; 281(5381): 13221326.
  20. 20. 王玉梅, 馮國(guó)和, 劉德剛, 等. Bcl2蛋白及caspase3基因表達(dá)與暴發(fā)性肝衰竭肝細(xì)胞凋亡關(guān)系的研究 [J]. 中國(guó)醫(yī)科大學(xué)學(xué)報(bào), 2006; 35(5): 494496.
  21. 21. 王玉梅, 馮國(guó)和, 黃芬, 等. 腫瘤壞死因子α及caspase3表達(dá)與暴發(fā)性肝衰竭細(xì)胞凋亡 [J]. 中華內(nèi)科雜志, 2003; 42(8): 566570.
  22. 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. 23. Fumarola C, Guidotti GG. Stressinduced apoptosis: toward a symmetry with receptormediated cell death [J]. Apoptosis, 2004; 9(1): 7782.
  24. 24. Meier P, Finch A, Evan G. Apoptosis in development[J]. Nature, 2000; 407(6805): 796801.
  25. 25. Gross GJ. Role of opioids in acute and delayed preconditioning [J]. J Mol Cell Cardiol, 2003; 35(7): 709718.