Yap介导瑞芬太尼预处理促进肝切除小鼠术后肝再生
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摘要
目的观察瑞芬太尼预处理对70%肝脏切除术诱导的小鼠肝再生的作用。方法选取6～8周龄C57雄性小鼠,随机分入对照组和瑞芬太尼组。称量小鼠的基础体重后,瑞芬太尼组小鼠于腹腔内注射瑞芬太尼30μg/kg,对照组注射等量0.9%氯化钠溶液,20～30 min后在异氟烷麻醉下行70%肝脏切除术。在肝切除术后6、24、36、48、120、192 h称量小鼠剩余30%肝湿重并计算肝体比,在肝切除术后24、36、48、120、192 h采用Ki-67免疫组织化学方法观察细胞增殖情况,在肝切除术后6、24、36、48、120 h采用实时定量PCR检测再生肝脏组织表皮生长因子受体(EGFR) mRNA表达水平,在肝切除术后120 h采用Western印迹法检测再生肝脏组织中Yap和磷酸化Yap(p-Yap)蛋白质表达水平。结果瑞芬太尼组肝切除术后48和120 h的肝体比均显著高于对照组同时间点(P值均<0.05)。瑞芬太尼组肝切除术后36、48、120、192 h的Ki-67阳性细胞率均显著高于对照组同时间点(P值分别<0.01、0.05)。瑞芬太尼组肝切除术后6 h的EGFR mRNA水平显著高于对照组同时间点(P<0.05)。瑞芬太尼组肝切除术后120 h的肝组织p-Yap蛋白质相对表达量显著低于对照组同时间点(P<0.05)。结论瑞芬太尼预处理可能通过促进Yap的功能性活化来促进70%肝脏切除后小鼠的肝再生。
Objective To explore the effect of remifentanil preconditioning on liver regeneration induced by 70% partial hepatectomy in mice. Methods C57 male mice, aged 6-8 weeks, were randomly divided into control group and remifentanil group. After weighing, the mice in remifentanil group were injected with remifentanil 30 μg/kg intraperitoneally, while the mice in normal saline group were injected with the same dose of normal saline. Then 70% partial hepatectomy was performed under isoflurane anesthesia within 20 to 30 min. Sampling and weighing the remaining 30% liver at 6, 24, 36, 48, 120 and 192 h after surgery to calculate the liver/body ratio. Immunohistochemical Ki-67 was used to detect cell proliferation at 24, 36, 48, 120 and 192 h after hepatectomy. mRNA levels of epidermal growth factor receptor(EGFR) in the liver were measured by real-time quantitative PCR(q-PCR) at 6, 24, 36, 48 and 120 h after hepatectomy. Yap and phosphorylated Yap(p-Yap) protein expression levels in the liver at 120 h after hepatectomy were detected with western blotting. Results The liver/body ratios of remifentanil group were significantly higher than those in control group at 48 and 120 h after hepatectomy(both P<0.05). The proportions of Ki-67 positive cells in remifentanil group were significantly higher than those in control group at 36, 48, 120 and 192 h after hepatectomy(P<0.01 or 0.05). The mRNA level of EGFR in remifentanil group was significantly higher than that in control group at 6 h after hepatectomy(P<0.05). The relative expression of p-Yap in regenerative liver of remifentanil group decreased significantly at 120 h after hepatectomy as compared to that in control group(P<0.05). Conclusion The preconditioning with remifentanil can promote liver regeneration after 70% partial hepatectomy in mice, which may be mediated by the activation of Yap.(Shanghai Med J, 2019, 42: 368-372)
Objective To explore the effect of remifentanil preconditioning on liver regeneration induced by 70% partial hepatectomy in mice. Methods C57 male mice, aged 6-8 weeks, were randomly divided into control group and remifentanil group. After weighing, the mice in remifentanil group were injected with remifentanil 30 μg/kg intraperitoneally, while the mice in normal saline group were injected with the same dose of normal saline. Then 70% partial hepatectomy was performed under isoflurane anesthesia within 20 to 30 min. Sampling and weighing the remaining 30% liver at 6, 24, 36, 48, 120 and 192 h after surgery to calculate the liver/body ratio. Immunohistochemical Ki-67 was used to detect cell proliferation at 24, 36, 48, 120 and 192 h after hepatectomy. mRNA levels of epidermal growth factor receptor(EGFR) in the liver were measured by real-time quantitative PCR(q-PCR) at 6, 24, 36, 48 and 120 h after hepatectomy. Yap and phosphorylated Yap(p-Yap) protein expression levels in the liver at 120 h after hepatectomy were detected with western blotting. Results The liver/body ratios of remifentanil group were significantly higher than those in control group at 48 and 120 h after hepatectomy(both P<0.05). The proportions of Ki-67 positive cells in remifentanil group were significantly higher than those in control group at 36, 48, 120 and 192 h after hepatectomy(P<0.01 or 0.05). The mRNA level of EGFR in remifentanil group was significantly higher than that in control group at 6 h after hepatectomy(P<0.05). The relative expression of p-Yap in regenerative liver of remifentanil group decreased significantly at 120 h after hepatectomy as compared to that in control group(P<0.05). Conclusion The preconditioning with remifentanil can promote liver regeneration after 70% partial hepatectomy in mice, which may be mediated by the activation of Yap.(Shanghai Med J, 2019, 42: 368-372)
引文
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[15] BAIER P,WOLF-VORBECK G,HEMPEL S,et al.Effect of liver regeneration after partial hepatectomy and ischemia-reperfusion on expression of growth factor receptors[J].World J Gastroenterol,2006,12(24):3835-3840.
[16] ZHAO B,WEI X,LI W,et al.Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control[J].Genes Dev,2007,21(21):2747-2761.
[17] CAMARGO F D,GOKHALE S,JOHNNIDIS J B,et al.YAP1 increases organ size and expands undifferentiated progenitor cells[J].Curr Biol,2007,17(23):2054-2060.
[18] GRIJALVA J L,HUIZENGA M,MUELLER K,et al.Dynamic alterations in Hippo signaling pathway and YAP activation during liver regeneration[J].Am J Physiol Gastrointest Liver Physiol,2014,307(2):G196-G204.
[2] ZHAO G,SHEN X,NAN H,et al.Remifentanil protects liver against ischemia/reperfusion injury through activation of anti-apoptotic pathways[J].J Surg Res,2013,183(2):827-834.
[3] JIANG Q,SONG X,CHEN Z,et al.Effects of remifentanil on hemodynamics,liver function and ICAM-1 expression in liver cancer patients undergoing surgery[J].Oncol Lett,2017,14(1):872-876.
[4] ZHU Y,WANG Y,DU B,et al.Could remifentanil reduce duration of mechanical ventilation in comparison with other opioids for mechanically ventilated patients?A systematic review and meta-analysis[J/OL].Crit Care,2017,21(1):206[2017-08-03].https://doi.org/10.1186/s13054-017-1789-8.
[5] YANG L Q,TAO K M,LIU Y T,et al.Remifentanil preconditioning reduces hepatic ischemia-reperfusion injury in rats via inducible nitric oxide synthase expression[J].Anesthesiology,2011,114(5):1036-1047.
[6] YANG Y,CHEN C,CUI C,et al.Indispensable role of β-arrestin2 in the protection of remifentanil preconditioning against hepatic ischemic reperfusion injury[J/OL].Sci Rep,2019,9(1):2087[2019-02-14].https://www.nature.com/articles/s41598-018-38456-9.
[7] SHEN J T,LI Y S,XIA Z Q,et al.Remifentanil preconditioning protects the small intestine against ischemia/reperfusion injury via intestinal δ- and μ-opioid receptors[J].Surgery,2016,159(2):548-559.
[8] HOU J,WANG H,LI X,et al.Remifentanil functions in the adaptive protection of cardiac function following ischemia[J].Exp Ther Med,2017,13(4):1514-1520.
[9] 方舒东,朱也森,徐辉,等.瑞芬太尼对大鼠全脑缺血再灌注后神经元凋亡和半胱天冬酶-3表达的影响[J].上海医学,2010,33(2):123-127,后插1.
[10] CUI C,YU F,YIN S,et al.Remifentanil preconditioning attenuates hepatic ischemia-reperfusion injury in rats via neuronal activation in dorsal vagal complex[J/OL].Mediators Inflamm,2018,2018:3260256[2018-05-10].https://doi.org/10.1155/2018/3260256.
[11] MAO S A,GLORIOSO J M,NYBERG S L.Liver regeneration[J].Transl Res,2014,163(4):352-362.
[12] MICHALOPOULOS G K,KHAN Z.Liver regeneration,growth factors,and amphiregulin[J].Gastroenterology,2005,128(2):503-506.
[13] HONG F,NGUYEN V A,SHEN X,et al.Rapid activation of protein kinase B/Akt has a key role in antiapoptotic signaling during liver regeneration[J].Biochem Biophys Res Commun,2000,279(3):974-979.
[14] NATARAJAN A,WAGNER B,SIBILIA M.The EGF receptor is required for efficient liver regeneration[J].Proc Natl Acad Sci U S A,2007,104(43):17081-17086.
[15] BAIER P,WOLF-VORBECK G,HEMPEL S,et al.Effect of liver regeneration after partial hepatectomy and ischemia-reperfusion on expression of growth factor receptors[J].World J Gastroenterol,2006,12(24):3835-3840.
[16] ZHAO B,WEI X,LI W,et al.Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control[J].Genes Dev,2007,21(21):2747-2761.
[17] CAMARGO F D,GOKHALE S,JOHNNIDIS J B,et al.YAP1 increases organ size and expands undifferentiated progenitor cells[J].Curr Biol,2007,17(23):2054-2060.
[18] GRIJALVA J L,HUIZENGA M,MUELLER K,et al.Dynamic alterations in Hippo signaling pathway and YAP activation during liver regeneration[J].Am J Physiol Gastrointest Liver Physiol,2014,307(2):G196-G204.
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