不同引流方式对梗阻性黄疸大鼠部分肝切除术后肝再生的影响

Effects of different biliary drainage methods on liver regeneration of obstructive jaundice rats after partial hepatectomy

  • 摘要: 目的:探讨不同引流方式对梗阻性黄疸大鼠部分肝切除术后肝再生的影响。
    方法: 建立梗阻性 黄疸70%部分肝切除SD大鼠模型。随后将120只SD大鼠按照随机数字表法分为对照组:行肝中、左叶切除;内引流组:于扩张胆管和十二指肠间置管引流;外引流组:于扩张胆管置管,导管另一端从腹腔引出。每组40只大鼠。内引流组和外引流组引流7 d后行肝中、左叶切除,于术后0、1、2、4、12、24、48、72 h收集3组大鼠血液及肝脏组织标本,测定肝再生率、有丝分裂指数。采用免疫组织化学染色法观察肝脏组织增殖细胞核抗原(PCNA)及信号传导与转录激活因子3(STAT3)的表达,ELISA法检测血清TNFα、IL6水平,RT PCR测定肝脏组织TNF α mRNA和IL6 mRNA的表达。组间比较采用单因素方差分析,两两比较采用 SNK 检验。
    结果:部分肝切除术后72 h内引流组SD大鼠肝再生率为94.86% ±3.7%。内引流组STAT3表达峰值显著高于对照组和外引流组( P <0.05)。内引流组TNF α和IL 〖KG*9〗6水平均于术后12 h达到高峰,分别为(227±23)U/L和(256±32)U/L;对照组和外引流组TNF α和IL 6水平均于术后24 h达到高峰,分别为(309±41)U/L和(388±40)U/L、(287±30)U/L和(346±33)U/L,内引流组术后0、1、2、4、12、24、48、72 h TNFα和IL 〖KG*9〗6水平显著低于相同时相点对照组和外引流组( P <0.05)。对照组、内引流组和外引流组大鼠肝组织TNF α mRNA 表达均于术后4 h达到高峰,分别为0.92±0.14、0.39±0.05、0.80±0.15,IL 6 mRNA于术后12 h达到高峰,分别为0.79±0.07、0.38±0.06、0.63±0.10,内引流组术后0、1、2、4、12、24、48、72 h TNF α mRNA和IL 6 mRNA表达显著低于相同时相点对照组和外引流组( P<0.05)。
    结论:内、外引流均可改善梗阻性黄疸大鼠剩余肝脏的再生能力,但内引流效果更明显。内引流术可能通过降低TNF α和IL6水平,影响STAT3表达而改善梗阻性黄疸大鼠剩余肝脏的再生能力。

     

    Abstract: Objective:To investigate the effects of internal and external biliary drainage on liver regeneration of the obstructive jaundice rats after partial hepatectomy.
    Methods:The rat models of obstructive jaundice with 70% liver resection were successfully constructed. All the 120 rats were randomly divided into the control group: rats received middle and left hepatic lobectomy; internal drainage group: a drainage tube was placed between the dilated bile duct and the duodenum; external drainage group: a drainage tube was placed in the dilated bile duct. There were 40 rats in each group. Rats in the internal and external drainage groups received middle and left hepatic lobectomy at postoperative day 7. The blood and hepatic tissues were collected at postoperative day 0, 1, 2, 4, 12, 24, 48, 72 hours after operation, and the rate of liver regeneration and mitotic index were determined. The expression of proliferating cell nuclear antigen (PCNA) and signal transducer and activator of transcription 3 (STAT3) in the hepatic tissues were detected by immunohistochemistry, and the levels of tumor necrosis factor α (TNF α) and interleukin6 (IL6) were detected by ELISA, and the mRNA expressions of TNF α and IL6 were detected by RT PCR. All data were analyzed using the one way analysis of variance or SNK test.
    Results:Within 72 hours after partial hepatectomy, the rate of liver regeneration of the internal drainage group was 94.86%±12.72%, which was significant higher than 62.39%±8.01% of the external drainage group and 45.77%±5.41% of the control group ( F=33.62, P <0.05). The mitotoic index and PCNA levels of the 3 groups had obvious increase at postoperative hour 12, and the mitotoic index and PCNA levels of the external drainage group reached peak at postoperative hour 24, which were 24.47%±4.01% and 88.1%±9.2%, respectively. the mitotoic index and PCNA levels of the control group and the external drainage group reached peak at postoperative hour 48, which were 15.80%±1.08%, 58.3%±5.8% and 18.40%± 1.12%, 70.2%±6.9%, respectively. The mitotoic index and PCNA levels of the internal drainage group were significantly higher than those of the control group and the external drainage group ( P <0.05). The expression of STAT3 expression of the internal drainage group reached peak at postoperative hour 4, which was 42.6%±3.6%; the expression of STAT3 expression of the control group and the external drainage group reached peak at postoperative hour 12, which were 22.9%±2.0% and 29.2%±3.7%. The peak level of STAT3 of the internal drainage group was significantly higher than those of the control group and the external drainage group ( P<0.05). The levels of TNF α and IL 6 of the internal drainage group reached peak at postoperative hour 12, which were (227±23)U/L and (256±32)U/L; the levels of TNF α and IL 6 of the control group and the external drainage group reached peak at postoperative hour 24, which were (309±41)U/L and (388±40)U/L, (287±30)U/L and (346±33)U/L, respectively. The levels of TNF α and IL 6 of the internal drainage group at postoperative hour 0, 1, 2, 4, 12, 24, 48, 72 were significantly lower than those of the control group and the external drainage group ( P <0.05). The expressions of TNF α mRNA of the control group, internal drainage group and external drainage group reached peak at postoperative hour 4, which were 0.92±0.14, 0.39±0.05, 0.80±0.15, respectively. The expressions of IL6 mRNA reached peak at postoperative hour 12, which were 0.79±0.07, 0.38±0.06, 0.63±0.10, respectively. The expressions of TNF α mRNA and IL 6 mRNA of the internal drainage group at postoperative hour 0, 1, 2, 4, 12, 24, 48, 72 were significantly lower than those of the control group and the external drainage group ( P<0.05).
    Conclusions:Both internal and external drainage can improve liver regeneration of obstructive jaundice rats following partial hepatectomy, while the effect of internal drainage is superior. Internal biliary drainage has influence on the expression of STAT3 by decreasing the levels of TNF α and IL 6, and help to improve liver regeneration of obstructive jaundice rats following partial hepatectomy.

     

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