ALPPS一期术后肝内干/祖细胞的变化及其与肝再生的关系
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摘要
目的:探讨肝内干/祖细胞在联合肝脏分割和门静脉结扎二步肝切除术(ALPPS)一期手术后肝再生中的作用。方法:将72只SD大鼠随机均分为ALPPS组、门静脉结扎(PVL)组和假手术组,分别行ALPPS一期手术、单纯PVL和假手术。分别在术后1、2、3、7 d检测各组血清转氨酶、炎症因子水平与肝右中叶肝再生率(HRR),并检测肝脏组织中细胞增殖指标Ki-67与肝内卵圆细胞(干/祖细胞)标志物OV-6表达水平。结果:与假手术组比较,ALPPS组与PVL组术后1~2 d的转氨酶与炎症因子水平均明显升高,且在ALPPS组的升高水平均大于PVL组(均P<0.05);ALPPS组与PVL组术后肝右中叶HRR及肝组织Ki-67阳性率明显升高,但ALPPS组在术后3、7 d的HRR明显高于PVL组,术后2、3 d的Ki-67阳性率明显高于PVL组(均P<0.05);ALPPS组与PVL组术后肝组织均有明显OV-6表达,但ALPPS组术后2、3 d的OV-6表达水平明显高于PVL组(均P<0.05)。结论:ALPPS一期手术诱导的肝再生明显优于PVL,机制可能为ALPPS术后较高的炎症状态使激活肝内干/祖细胞的动员和活化,从而促进快速肝再生有关。
Objective: To investigate the role of intrahepatic stem/progenitor cells in liver regeneration after the first stage of associating liver partition and portal vein ligation for staged hepatectomy(ALPPS). Methods: Seventy-two SD rats were equally randomized into ALPPS group, portal vein ligation(PVL) group and sham operation group, and then underwent the first stage of ALPPS, PVL alone and sham operation, respectively. The serum levels of transaminases and inflammatory factors and the hepatic regeneration rate(HRR) of the right middle lobe of the liver were determined, and the expressions of cell proliferation index Ki-67 and OV-6, the biomarker of oval cells(stem/progenitor cells), were also examined in each group of rats at postoperative day(POD) 1, 2, 3 and 7, respectively.Results: Compared with sham operation group, the serum levels of transaminases and inflammatory factors were significantly increased in both ALPPS group and PVL group on POD 1 to 2, and their increasing amplitudes in ALPPS group were all significantly higher than those in PVL group(all P<0.05); the HRR of the right middle lobe of the liver and positive Ki-67 rate in the liver tissue were significantly increases in both ALPPS group and PVL group, but the HRR was significantly higher on POD 3 and 7, the positive Ki-67 rate was significantly higher on POD 2 and 3 in ALPPS group than those in PVL group(all P<0.05); obvious OV-6 expression was seen in either ALPPS group or PVL group, but its expression levels in ALPPS group were significantly higher than those in PVL group on POD 2 and 3(both P<0.05).Conclusion: The first stage of ALPPS is superior to single PVL for inducing liver regeneration, and the mechanism is probably related to higher inflammatory status after ALPPS which may cause the mobilization and activation of the intrahepatic stem/progenitor cells, and thereby promote the rapid liver regeneration.
Objective: To investigate the role of intrahepatic stem/progenitor cells in liver regeneration after the first stage of associating liver partition and portal vein ligation for staged hepatectomy(ALPPS). Methods: Seventy-two SD rats were equally randomized into ALPPS group, portal vein ligation(PVL) group and sham operation group, and then underwent the first stage of ALPPS, PVL alone and sham operation, respectively. The serum levels of transaminases and inflammatory factors and the hepatic regeneration rate(HRR) of the right middle lobe of the liver were determined, and the expressions of cell proliferation index Ki-67 and OV-6, the biomarker of oval cells(stem/progenitor cells), were also examined in each group of rats at postoperative day(POD) 1, 2, 3 and 7, respectively.Results: Compared with sham operation group, the serum levels of transaminases and inflammatory factors were significantly increased in both ALPPS group and PVL group on POD 1 to 2, and their increasing amplitudes in ALPPS group were all significantly higher than those in PVL group(all P<0.05); the HRR of the right middle lobe of the liver and positive Ki-67 rate in the liver tissue were significantly increases in both ALPPS group and PVL group, but the HRR was significantly higher on POD 3 and 7, the positive Ki-67 rate was significantly higher on POD 2 and 3 in ALPPS group than those in PVL group(all P<0.05); obvious OV-6 expression was seen in either ALPPS group or PVL group, but its expression levels in ALPPS group were significantly higher than those in PVL group on POD 2 and 3(both P<0.05).Conclusion: The first stage of ALPPS is superior to single PVL for inducing liver regeneration, and the mechanism is probably related to higher inflammatory status after ALPPS which may cause the mobilization and activation of the intrahepatic stem/progenitor cells, and thereby promote the rapid liver regeneration.
引文
[1]Agrawal S,Belghiti J.Oncologic resection for malignant tumors of the liver[J].Ann Surg,2011,253(4):656-665.doi:10.1097/SLA.0b013e3181fc08ca.
[2]Clavien PA,Petrowsky H,DeOliveira ML,et al.Strategies for safer liver surgery and partial liver transplantation[J].N Engl J Med,2007,356(15):1545-1559.
[3]Robles R,Parrilla P,López-Conesa A,et al.Tourniquet modification of the associating liver partition and portal ligation for staged hepatectomy procedure[J].Br J Surg,2014,101(9):1129-1134.doi:10.1002/bjs.9547.
[4]Schnitzbauer AA,Lang SA,Goessmann H,et al.Right portal vein ligation combined with in situ splitting induces rapid left lateral liver lobe hypertrophy enabling 2-staged extended right hepatic resection in small-for-size settings[J].Ann Surg,2012,255(3):405-414.doi:10.1097/SLA.0b013e31824856f5.
[5]Andersen KJ,Knudsen AR,Jepsen BN,et al.A new technique for accelerated liver regeneration:An experimental study in rats[J].Surgery,2017,162(2):233-247.doi:10.1016/j.surg.2017.03.002.
[6]Schadde E,Ardiles V,Slankamenac K,et al.ALPPS offers a better chance of complete resection in patients with primarily unresectable liver tumors compared with conventional-staged hepatectomies:results of a multicenter analysis[J].World J Surg,2014,38(6):1510-1519.doi:10.1007/s00268-014-2513-3.
[7]Lang SA,Loss M,Benseler V,et al.Long-term results after insitu split(ISS)liver resection[J].Langenbecks Arch Surg,2015,400(3):361-369.doi:10.1007/s00423-015-1285-z.
[8]Liu WH,Ren LN,Wang T,et al.The Involving Roles of Intrahepatic and Extrahepatic Stem/Progenitor Cells(SPCs)to Liver Regeneration[J].Int J Biol Sci,2016,12(8):954-963.doi:10.7150/ijbs.15715.
[9]黄民,黄粲宸,刘卫辉.肝内外干/祖细胞参与肝再生的研究进展[J].中国普通外科杂志,2017,26(7):926-933.doi:10.3978/j.issn.1005-6947.2017.07.018.Huang M,Huang CC,Liu WH.Participation of intra-and extrahepatic stem/progenitor cells in liver regeneration:recent advances[J].Chinese Journal of General Surgery,2017,26(7):926-933.doi:10.3978/j.issn.1005-6947.2017.07.018.
[10]Van Haele M,Roskams T.Hepatic Progenitor Cells:An Update[J].Gastroenterol Clin North Am,2017,46(2):409-420.doi:10.1016/j.gtc.2017.01.011.
[11]Miyajima A,Tanaka M,Itoh T.Stem/progenitor cells in liver development,homeostasis,regeneration,and reprogramming[J].Cell Stem Cell,2014,14(5):561-574.doi:10.1016/j.stem.2014.04.010.
[12]Liu WH,Song FQ,Ren LN,et al.The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases[J].J Cell Mol Med,2015,19(3):511-520.doi:10.1111/jcmm.12482.
[13]Itoh T,Miyajima A.Liver regeneration by stem/progenitor cells[J].Hepatology,2014,59(4):1617-1626.doi:10.1002/hep.26753.
[14]刘伟伟,刘洪,余锋,等.联合肝脏分割和门静脉结扎二步肝切除术大鼠模型的建立[J].中国普通外科杂志,2017,26(1):50-56.doi:10.3978/j.issn.1005-6947.2017.01.009.Liu WW,Liu H,Yu F,et al.Establishment of rat model of associating liver partition and portal vein ligation for staged hepatectomy[J].Chinese Journal of General Surgery,2017,26(1):50-56.doi:10.3978/j.issn.1005-6947
[15]顾向前,张鲁洲,曹顺琪,等.联合肝脏分割和门静脉结扎的分期肝切除术大鼠模型的建立[J].中华肝胆外科杂志,2015,21(12):836-839.doi:10.3760/cma.j.issn.1007-8118.2015.12.013.Gu XQ,Zhang LZ,Cao SQ,et al.Establishing rat model of associating liver partition with portal vein ligation for staged hepatectomy(ALPPS)[J].Chinese Journal of Hepatobiliary Surgery,2015,21(12):836-839.doi:10.3760/cma.j.issn.1007-8118.2015.12.013.
[16]Wei W,Zhang T,Zafarnia S,et al.Establishment of a rat model:Associating liver partition with portal vein ligation for staged hepatectomy[J].Surgery,2016,159(5):1299-1307.doi:10.1016/j.surg.2015.12.005.
[17]Yao L,Li C,Ge X,et al.Establishment of a rat model of portal vein ligation combined with in situ splitting[J].PLoS One,2014,9(8):e105511.doi:10.1371/journal.pone.0105511.
[18]Golriz M1,Majlesara A1,El Sakka S1,et al.Small for Size and Flow(SFSF)syndrome:An alternative description for posthepatectomy liver failure[J].Clin Res Hepatol Gastroenterol,2016,40(3):267-275.doi:10.1016/j.clinre.2015.06.024.
[19]Zhang GQ,Zhang ZW,Lau WY,et al.Associating liver partition and portal vein ligation for staged hepatectomy(ALPPS):a new strategy to increase resectability in liver surgery[J].Int J Surg,2014,12(5):437-441.doi:10.1016/j.ijsu.2014.03.009.
[20]Dezs?K,Papp V,Bugyik E,et al.Structural analysis of ovalcellmediated liver regeneration in rats[J].Hepatology,2012,56(4):1457-1467.doi:10.1002/hep.25713.
[21]Guo J,Li J,Lu Y,et al.A novel technique for hepatic progenitor cell isolation from normal adult rat livers[J].ASAIO J,2012,58(1):73-78.doi:10.1097/MAT.0b013e318239fce5.
[22]Schmidt-Arras D,Rose-John S.IL-6 pathway in the liver:From physiopathology to therapy[J].J Hepatol,2016,64(6):1403-1415.doi:10.1016/j.jhep.2016.02.004.
[23]Wang S,Lee JS,Hyun J,et al.Tumor necrosis factor-inducible gene6 promotes liver regeneration in mice with acute liver injury[J].Stem Cell Res Ther,2015,6:20.doi:10.1186/s13287-015-0019-z.
[24]Li Z,Chen J,Li L,et al.In vitro and in vivo characteristics of hepatic oval cells modified with human hepatocyte growth factor[J].Cell Mol Biol Lett,2013,18(4):507-521.doi:10.2478/s11658-013-0104-1
[25]Ji T,Li G,Chen J,et al.Distinct role of interleukin-6 and tumor necrosis factor receptor-1 in oval cell-mediated liver regeneration and inflammation-associated hepatocarcinogenesis[J].Oncotarget,2016,7(41):66635-66646.doi:10.18632/oncotarget.11365.
[26]Knight B,Yeoh GC,Husk KL,et al.Impaired preneoplastic changes and liver tumor formation in tumor necrosis factor receptor type 1knockout mice[J].J Exp Med,2000,192(12):1809-1818.
[27]Matthews VB,Klinken E,Yeoh GC.Direct effects of interleukin-6on liver progenitor oval cells in culture[J].Wound Repair Regen,2004,12(6):650-656.
[2]Clavien PA,Petrowsky H,DeOliveira ML,et al.Strategies for safer liver surgery and partial liver transplantation[J].N Engl J Med,2007,356(15):1545-1559.
[3]Robles R,Parrilla P,López-Conesa A,et al.Tourniquet modification of the associating liver partition and portal ligation for staged hepatectomy procedure[J].Br J Surg,2014,101(9):1129-1134.doi:10.1002/bjs.9547.
[4]Schnitzbauer AA,Lang SA,Goessmann H,et al.Right portal vein ligation combined with in situ splitting induces rapid left lateral liver lobe hypertrophy enabling 2-staged extended right hepatic resection in small-for-size settings[J].Ann Surg,2012,255(3):405-414.doi:10.1097/SLA.0b013e31824856f5.
[5]Andersen KJ,Knudsen AR,Jepsen BN,et al.A new technique for accelerated liver regeneration:An experimental study in rats[J].Surgery,2017,162(2):233-247.doi:10.1016/j.surg.2017.03.002.
[6]Schadde E,Ardiles V,Slankamenac K,et al.ALPPS offers a better chance of complete resection in patients with primarily unresectable liver tumors compared with conventional-staged hepatectomies:results of a multicenter analysis[J].World J Surg,2014,38(6):1510-1519.doi:10.1007/s00268-014-2513-3.
[7]Lang SA,Loss M,Benseler V,et al.Long-term results after insitu split(ISS)liver resection[J].Langenbecks Arch Surg,2015,400(3):361-369.doi:10.1007/s00423-015-1285-z.
[8]Liu WH,Ren LN,Wang T,et al.The Involving Roles of Intrahepatic and Extrahepatic Stem/Progenitor Cells(SPCs)to Liver Regeneration[J].Int J Biol Sci,2016,12(8):954-963.doi:10.7150/ijbs.15715.
[9]黄民,黄粲宸,刘卫辉.肝内外干/祖细胞参与肝再生的研究进展[J].中国普通外科杂志,2017,26(7):926-933.doi:10.3978/j.issn.1005-6947.2017.07.018.Huang M,Huang CC,Liu WH.Participation of intra-and extrahepatic stem/progenitor cells in liver regeneration:recent advances[J].Chinese Journal of General Surgery,2017,26(7):926-933.doi:10.3978/j.issn.1005-6947.2017.07.018.
[10]Van Haele M,Roskams T.Hepatic Progenitor Cells:An Update[J].Gastroenterol Clin North Am,2017,46(2):409-420.doi:10.1016/j.gtc.2017.01.011.
[11]Miyajima A,Tanaka M,Itoh T.Stem/progenitor cells in liver development,homeostasis,regeneration,and reprogramming[J].Cell Stem Cell,2014,14(5):561-574.doi:10.1016/j.stem.2014.04.010.
[12]Liu WH,Song FQ,Ren LN,et al.The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases[J].J Cell Mol Med,2015,19(3):511-520.doi:10.1111/jcmm.12482.
[13]Itoh T,Miyajima A.Liver regeneration by stem/progenitor cells[J].Hepatology,2014,59(4):1617-1626.doi:10.1002/hep.26753.
[14]刘伟伟,刘洪,余锋,等.联合肝脏分割和门静脉结扎二步肝切除术大鼠模型的建立[J].中国普通外科杂志,2017,26(1):50-56.doi:10.3978/j.issn.1005-6947.2017.01.009.Liu WW,Liu H,Yu F,et al.Establishment of rat model of associating liver partition and portal vein ligation for staged hepatectomy[J].Chinese Journal of General Surgery,2017,26(1):50-56.doi:10.3978/j.issn.1005-6947
[15]顾向前,张鲁洲,曹顺琪,等.联合肝脏分割和门静脉结扎的分期肝切除术大鼠模型的建立[J].中华肝胆外科杂志,2015,21(12):836-839.doi:10.3760/cma.j.issn.1007-8118.2015.12.013.Gu XQ,Zhang LZ,Cao SQ,et al.Establishing rat model of associating liver partition with portal vein ligation for staged hepatectomy(ALPPS)[J].Chinese Journal of Hepatobiliary Surgery,2015,21(12):836-839.doi:10.3760/cma.j.issn.1007-8118.2015.12.013.
[16]Wei W,Zhang T,Zafarnia S,et al.Establishment of a rat model:Associating liver partition with portal vein ligation for staged hepatectomy[J].Surgery,2016,159(5):1299-1307.doi:10.1016/j.surg.2015.12.005.
[17]Yao L,Li C,Ge X,et al.Establishment of a rat model of portal vein ligation combined with in situ splitting[J].PLoS One,2014,9(8):e105511.doi:10.1371/journal.pone.0105511.
[18]Golriz M1,Majlesara A1,El Sakka S1,et al.Small for Size and Flow(SFSF)syndrome:An alternative description for posthepatectomy liver failure[J].Clin Res Hepatol Gastroenterol,2016,40(3):267-275.doi:10.1016/j.clinre.2015.06.024.
[19]Zhang GQ,Zhang ZW,Lau WY,et al.Associating liver partition and portal vein ligation for staged hepatectomy(ALPPS):a new strategy to increase resectability in liver surgery[J].Int J Surg,2014,12(5):437-441.doi:10.1016/j.ijsu.2014.03.009.
[20]Dezs?K,Papp V,Bugyik E,et al.Structural analysis of ovalcellmediated liver regeneration in rats[J].Hepatology,2012,56(4):1457-1467.doi:10.1002/hep.25713.
[21]Guo J,Li J,Lu Y,et al.A novel technique for hepatic progenitor cell isolation from normal adult rat livers[J].ASAIO J,2012,58(1):73-78.doi:10.1097/MAT.0b013e318239fce5.
[22]Schmidt-Arras D,Rose-John S.IL-6 pathway in the liver:From physiopathology to therapy[J].J Hepatol,2016,64(6):1403-1415.doi:10.1016/j.jhep.2016.02.004.
[23]Wang S,Lee JS,Hyun J,et al.Tumor necrosis factor-inducible gene6 promotes liver regeneration in mice with acute liver injury[J].Stem Cell Res Ther,2015,6:20.doi:10.1186/s13287-015-0019-z.
[24]Li Z,Chen J,Li L,et al.In vitro and in vivo characteristics of hepatic oval cells modified with human hepatocyte growth factor[J].Cell Mol Biol Lett,2013,18(4):507-521.doi:10.2478/s11658-013-0104-1
[25]Ji T,Li G,Chen J,et al.Distinct role of interleukin-6 and tumor necrosis factor receptor-1 in oval cell-mediated liver regeneration and inflammation-associated hepatocarcinogenesis[J].Oncotarget,2016,7(41):66635-66646.doi:10.18632/oncotarget.11365.
[26]Knight B,Yeoh GC,Husk KL,et al.Impaired preneoplastic changes and liver tumor formation in tumor necrosis factor receptor type 1knockout mice[J].J Exp Med,2000,192(12):1809-1818.
[27]Matthews VB,Klinken E,Yeoh GC.Direct effects of interleukin-6on liver progenitor oval cells in culture[J].Wound Repair Regen,2004,12(6):650-656.