IL-6/STAT3信号通路与小鼠联合原位肝劈开和门静脉分支结扎术后肝脏再生的关系
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摘要
目的:建立联合原位肝劈开和门静脉分支结扎(associating liver partition and portal vein ligation for staged heptectomy,ALPPS)小鼠模型,研究IL-6/STAT3信号通路与ALPPS促进肝脏再生的关系。方法:将45只BALB/C小鼠随机分为3组,分别为ALPPS组、门静脉分支结扎(portal vein ligation,PVL)组和假手术(SHAM)组,每组15只。PVL组即结扎支配左外叶、中叶右侧和右叶的门静脉分支,保留支配中叶左侧和尾状叶的门静脉分支血流。ALPPS组即在PVL组的基础上,沿着中叶缺血线将中叶离断。SHAM组即离断肝韧带,游离肝脏各叶。通过测定血浆ALT值,评估ALLPS和PVL术后肝功能情况。用肝脏再生指数评估肝脏再生情况。免疫组化检测Ki-67阳性蛋白表达来评估肝细胞增殖情况。免疫印迹技术被用来检测肝组织IL-6/STAT3信号通路变化情况。结果:术后48 h的ALPPS组ALT值高于PVL组,且两者有统计学差异(P=0.000)。而ALPPS和PVL术后96 h的ALT值较48 h下降,且两者在术后96 h的ALT值无明显统计学差异(P=0.094)。术后48 h和96 h的ALPPS组的肝脏再生指数(P48 h=0.000,P96 h=0.000)和Ki-67阳性细胞与肝细胞总数比值(P48 h=0.001,P96 h=0.000)高于PVL组,且两者有统计学差异。术后48 h的ALPPS组IL-6蛋白的表达和STAT3蛋白的磷酸化程度高于SHAM组(PIL-6=0.000, Pp STAT3=0.000)和PVL组(PIL-6=0.011,Pp STAT3=0.003),且有统计学差异。结论:成功建立了小鼠ALPPS模型,ALPPS比PVL更能促进肝脏再生和IL-6/STAT3信号通路与ALPPS介导肝脏再生的过程存在密切的关系。
Objective:To establish a mouse model of associating liver partition and portal vein ligation for staged hepatectomy(ALPPS),and to investigate the association of the interleukin-6(IL-6)/STAT3 signaling pathway with liver regeneration promoted by ALPPS. Methods:A total of 45 BALB/C mice were randomly divided into ALPPS group,portal vein ligation(PVL)group,and shamoperated group(SHAM group),with 15 mice in each group. The PVL group was treated with ligation of the portal vein branches for the left lateral lobe,the right side of the middle lobe,and the right lobe while preserving the blood flow in the portal vein branches for the left side of the middle lobe and the caudate lobe. The ALPPS group was treated with partition of the middle lobe along the demarcation line in addition to the treatment in the PVL group. The SHAM group was treated by dissociating the liver lobes along the hepatic ligaments. The plasma level of alanine aminotransferase(ALT)was measured to evaluate liver function after ALPPS and PVL. The liver regeneration index was used to evaluate liver regeneration. The expression of Ki-67 protein was measured by immunohistochemistry to evaluate the proliferation of hepatocytes. Western blotting was used to examine the change in the IL-6/STAT3 signaling pathway in liver tissue. Results:At 48 hours after surgery,the ALPPS group had a significantly higher ALT level than the PVL group(P=0.000);from 48 to 96 hours after surgery,both ALPPS and PVL groups had a reduction in ALT level,and there was no significant difference in ALT level between the ALPPS group and the PVL group at 96 hours after surgery(P=0.094). At 48 and 96 hours after surgery,the ALPPS had significantly higher liver regeneration index(P48 h=0.000,P96 h=0.000)and percentage of Ki-67-positive hepatocytes(P48 h=0.001,P96 h=0.000)than the PVL group. At 48 hours after surgery,the ALPPS group had significantly higher expression of IL-6 protein and phosphorylation of STAT3 protein than the SHAM group(PIL-6=0.000,Pp STAT3=0.000)and the PVL group(PIL-6=0.011,Pp STAT3=0.003). Conclusion:A mouse model of ALPPS was successfully established. Compared with PVL,ALPPS can better promote liver regeneration and the IL-6/STAT3 signaling pathway and is closely associated with ALPSS-mediated liver regeneration in mice.
Objective:To establish a mouse model of associating liver partition and portal vein ligation for staged hepatectomy(ALPPS),and to investigate the association of the interleukin-6(IL-6)/STAT3 signaling pathway with liver regeneration promoted by ALPPS. Methods:A total of 45 BALB/C mice were randomly divided into ALPPS group,portal vein ligation(PVL)group,and shamoperated group(SHAM group),with 15 mice in each group. The PVL group was treated with ligation of the portal vein branches for the left lateral lobe,the right side of the middle lobe,and the right lobe while preserving the blood flow in the portal vein branches for the left side of the middle lobe and the caudate lobe. The ALPPS group was treated with partition of the middle lobe along the demarcation line in addition to the treatment in the PVL group. The SHAM group was treated by dissociating the liver lobes along the hepatic ligaments. The plasma level of alanine aminotransferase(ALT)was measured to evaluate liver function after ALPPS and PVL. The liver regeneration index was used to evaluate liver regeneration. The expression of Ki-67 protein was measured by immunohistochemistry to evaluate the proliferation of hepatocytes. Western blotting was used to examine the change in the IL-6/STAT3 signaling pathway in liver tissue. Results:At 48 hours after surgery,the ALPPS group had a significantly higher ALT level than the PVL group(P=0.000);from 48 to 96 hours after surgery,both ALPPS and PVL groups had a reduction in ALT level,and there was no significant difference in ALT level between the ALPPS group and the PVL group at 96 hours after surgery(P=0.094). At 48 and 96 hours after surgery,the ALPPS had significantly higher liver regeneration index(P48 h=0.000,P96 h=0.000)and percentage of Ki-67-positive hepatocytes(P48 h=0.001,P96 h=0.000)than the PVL group. At 48 hours after surgery,the ALPPS group had significantly higher expression of IL-6 protein and phosphorylation of STAT3 protein than the SHAM group(PIL-6=0.000,Pp STAT3=0.000)and the PVL group(PIL-6=0.011,Pp STAT3=0.003). Conclusion:A mouse model of ALPPS was successfully established. Compared with PVL,ALPPS can better promote liver regeneration and the IL-6/STAT3 signaling pathway and is closely associated with ALPSS-mediated liver regeneration in mice.
引文
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[12] Schlegel A,Lesurtel M,Melloul E,et al. ALPPS:from human to mice highlighting accelerated and novel mechanisms of liver regeneration[J]. Ann Surg,2014,260(5):839-846.
[13] Fausto N,Campbell JS,Riehle KJ. Liver regeneration[J]. Hepatology,2006,43(2S1):45-53.
[14] Cressman DE,Greenbaum LE,DeAngelis RA,et al. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice[J]. Science,1996,274(5291):1379-1383.
[15] Sakamoto T,Liu Z,Murase N,et al. Mitosis and apoptosis in the liver of interleukin-6-deficient mice after partial hepatectomy[J]. Hepatology,1999,29(2):403-411.
[16] Wüstefeld T,Rakemann T,Kubicka S,et al. Hyperstimulation with interleukin 6 inhibits cell cycle progression after hepatectomy in mice[J]. Hepatology,2000,32(3):514-522.
[17] Blindenbacher A,Wang X,Langer I,et al. Interleukin 6 is important for survival after partial hepatectomy in mice[J]. Hepatology,2003,38(3):674-682.
[18] Heinrich S,Jochum W,Graf R,et al. Portal vein ligation and partial hepatectomy differentially influence growth of intrahepatic metastasis and liver regeneration in mice[J]. J Hepatol,2006,45(1):35-42.
[19] Furrer K,Tian Y,Pfammatter T,et al. Selective portal vein embolization and ligation trigger different regenerative responses in the rat liver[J]. Hepatology,2008,47(5):1615-1623.
[20] Langiewicz M,Schlegel A,Saponara E,et al. Hedgehog pathway mediates early acceleration of liver regeneration induced by a novel two-staged hepatectomy in mice[J]. J Hepatol,2017,66(3):560-570.
[21] Lau WY,Lai EC. Modifications of ALPPS-from complex to more complex or from complex to less complex operations[J]. Hepatobiliary Pancreat Dis Int,2017,16(4):346-352.
[2] Bruix J,Gores GJ,Mazzaferro V. Hepatocellular carcinoma:clinical frontiers and perspectives[J]. Gut,2014,63(5):844-855.
[3] Ratti F,Schadde E,Masetti M,et al. Strategies to increase the resectability of patients with colorectal liver metastases:a multi-center case-match analysis of ALPPS and conventional two-stage hepatectomy[J]. Ann Surg Oncol,2015,22(6):1933-1942.
[4] Sandstrom P,Rsok BI,Sparrelid E,et al. ALPPS improves resectability compared with conventional two-stage hepatectomy in patients with advanced colorectal liver metastasis:results from a scandinavian multicenter randomized controlled trial(LIGRO trial)[J]. Ann Surg,2018,267(5):833-840.
[5] 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.
[6] Chia DKA,Yeo Z,Loh SEK,et al. Greater hypertrophy can be achieved with associating liver partition with portal vein ligation for staged hepatectomy compared to conventional staged hepatectomy,but with a higher price to pay?[J]. Am J Surg,2018,215(1):131-137.
[7] Hunter CA,Jones SA. IL-6 as a keystone cytokine in health and disease[J]. Nat Immunol,2015,16(5):448-457.
[8]Sansone P,Bromberg J. Targeting the interleukin-6/Jak/stat pathway in human malignancies[J]. J Clin Oncol,2012,30(9):1005-1014.
[9] 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.
[10] Zhou Z,Xu M,Lin N,et al. Associating liver partition and portal vein ligation for staged hepatectomy versus conventional two-stage hepatectomy:a systematic review and meta-analysis[J]. World J Surg Oncol,2017,15(1):227.
[11] Matsuo K,Murakami T,Kawaguchi D,et al. Histologic features after surgery associating liver partition and portal vein ligation for staged hepatectomy versus those after hepatectomy with portal vein embolization[J]. Surgery,2016,159(5):1289-1298.
[12] Schlegel A,Lesurtel M,Melloul E,et al. ALPPS:from human to mice highlighting accelerated and novel mechanisms of liver regeneration[J]. Ann Surg,2014,260(5):839-846.
[13] Fausto N,Campbell JS,Riehle KJ. Liver regeneration[J]. Hepatology,2006,43(2S1):45-53.
[14] Cressman DE,Greenbaum LE,DeAngelis RA,et al. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice[J]. Science,1996,274(5291):1379-1383.
[15] Sakamoto T,Liu Z,Murase N,et al. Mitosis and apoptosis in the liver of interleukin-6-deficient mice after partial hepatectomy[J]. Hepatology,1999,29(2):403-411.
[16] Wüstefeld T,Rakemann T,Kubicka S,et al. Hyperstimulation with interleukin 6 inhibits cell cycle progression after hepatectomy in mice[J]. Hepatology,2000,32(3):514-522.
[17] Blindenbacher A,Wang X,Langer I,et al. Interleukin 6 is important for survival after partial hepatectomy in mice[J]. Hepatology,2003,38(3):674-682.
[18] Heinrich S,Jochum W,Graf R,et al. Portal vein ligation and partial hepatectomy differentially influence growth of intrahepatic metastasis and liver regeneration in mice[J]. J Hepatol,2006,45(1):35-42.
[19] Furrer K,Tian Y,Pfammatter T,et al. Selective portal vein embolization and ligation trigger different regenerative responses in the rat liver[J]. Hepatology,2008,47(5):1615-1623.
[20] Langiewicz M,Schlegel A,Saponara E,et al. Hedgehog pathway mediates early acceleration of liver regeneration induced by a novel two-staged hepatectomy in mice[J]. J Hepatol,2017,66(3):560-570.
[21] Lau WY,Lai EC. Modifications of ALPPS-from complex to more complex or from complex to less complex operations[J]. Hepatobiliary Pancreat Dis Int,2017,16(4):346-352.