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nodosin促进大鼠部分肝移植肝细胞再生的实验研究

周昕 马铁祥 李丰 刘翔 曾剑 李绍杰 廖国庆

周昕, 马铁祥, 李丰, 等. nodosin促进大鼠部分肝移植肝细胞再生的实验研究[J]. 器官移植, 2016, 7(2): 132-138. doi: 10.3969/j.issn.1674-7445.2016.02.011
引用本文: 周昕, 马铁祥, 李丰, 等. nodosin促进大鼠部分肝移植肝细胞再生的实验研究[J]. 器官移植, 2016, 7(2): 132-138. doi: 10.3969/j.issn.1674-7445.2016.02.011
Zhou Xin, Ma Tiexiang, Li Feng, et al. Effect of nodosin on accelerating hepatocyte regeneration after partial liver transplantation in rats[J]. ORGAN TRANSPLANTATION, 2016, 7(2): 132-138. doi: 10.3969/j.issn.1674-7445.2016.02.011
Citation: Zhou Xin, Ma Tiexiang, Li Feng, et al. Effect of nodosin on accelerating hepatocyte regeneration after partial liver transplantation in rats[J]. ORGAN TRANSPLANTATION, 2016, 7(2): 132-138. doi: 10.3969/j.issn.1674-7445.2016.02.011

nodosin促进大鼠部分肝移植肝细胞再生的实验研究

doi: 10.3969/j.issn.1674-7445.2016.02.011
基金项目: 

湖南省科技厅计划项目 2012SK3190

详细信息
    通讯作者:

    周昕, Email:zhx0161@sina.com

  • 中图分类号: R617;R-332

Effect of nodosin on accelerating hepatocyte regeneration after partial liver transplantation in rats

More Information
    Corresponding author: Zhou Xin, Email:zhx0161@sina.com
  • 摘要:   目的  探讨溪黄草有效成分nodosin对大鼠减体积肝移植术后肝细胞再生的影响。  方法  以Wistar大鼠作为供体, SD大鼠作为受体, 采用改良的二袖套法建立大鼠部分肝移植模型, 将24只受体大鼠随机分为nodosin组和对照组。nodosin组于肝移植术后经尾静脉注射nodosin 100μg/ml。在术后3 d和7 d分别处死大鼠取外周血浆及肝脏组织标本。分光光度法检测外周血浆中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和白蛋白(ALB)含量; 光学显微镜下观察肝组织形态学改变; 采用免疫组织化学法检测肝组织增殖细胞核抗原(PCNA)抗体的表达水平; 采用蛋白印迹法(Western blot)检测肝组织中磷酸化蛋白激酶(p-AKT)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)、细胞周期素D1(cyclin D1)、血红素加氧酶(HO)-1蛋白的表达水平; 采用磷脂酰丝氨酸外翻分析(Annexin V法)和dUTP缺口末端标记(TUNEL)法检测肝细胞的凋亡情况。  结果  术后3 d和7 d, 与对照组比较, nodosin组大鼠血清中ALT、AST含量较低, ALB含量较高(均为P < 0.05), 同时nodosin能够减轻大鼠肝移植术后肝组织病理损伤。nodosin组的PCNA阳性表达细胞数明显多于对照组(P < 0.05)。nodosin组增殖相关蛋白p-AKT、p-mTOR、cyclin D1和HO-1蛋白表达水平均明显高于对照组(均为P < 0.05)。nodosin组发生凋亡的肝细胞数量和比例明显低于对照组(P < 0.05)。  结论  大鼠肝移植术后应用nodosin具有减少肝细胞凋亡数量, 促进肝细胞增殖的作用。

     

  • 图  1  两组大鼠术后3 d和7 d肝组织病理变化(HE,×200)

    注:A图为对照组术后3 d;B图为nodosin组术后3 d;C图为对照组术后7 d;D图为nodosin组术后7 d

    Figure  1.  Pathological changes of liver tissues of rats in two groups at 3 d and 7 d after operation

    图  2  两组大鼠术后3 d和7 d肝细胞PCNA阳性细胞数的比较

    注:A图为对照组术后3 d肝细胞PCNA的免疫组化染色切片(DAB,×100);B图为nodosin组术后3 d肝细胞PCNA的免疫组化染色切片(DAB,×100);C图为对照组术后7 d肝细胞PCNA的免疫组化染色切片(DAB,×100);D为nodosin组术后7 d肝细胞PCNA的免疫组化染色切片(DAB,×100);E图为两组PCNA蛋白阳性表达数量对比,与对照组相比,aP < 0.05

    Figure  2.  Comparison of positive cell count of PCNA in hepatocyte of rats in two groups at 3 d and 7 d after operation

    图  3  两组大鼠肝细胞中p-AKT、p-mTOR、cyclin D1和HO-1蛋白表达水平的变化

    Figure  3.  Alteration of protein expression levels of p-AKT, p-mTOR, cyclin D1 and HO-1 in hepatocyte of rats in two groups

    图  4  两组大鼠术后3 d和7 d肝细胞的凋亡情况

    注:A图为对照组术后3 d流式细胞图;B图为nodosin组术后3 d流式细胞图;C图为对照组术后7 d流式细胞图;D图为nodosin组术后7 d流式细胞图;E图为两组大鼠肝细胞凋亡比例的比较,与对照组相比,aP < 0.05;F图为对照组术后3 d肝细胞凋亡情况(TUNEL,×100);G图为nodosin组术后3 d肝细胞凋亡情况(TUNEL,×100);H图为对照组术后7 d肝细胞凋亡情况(TUNEL,×100);I图为nodosin组术后7 d肝细胞凋亡情况(TUNEL,×100);J图为两组大鼠肝细胞凋亡指数的比较,与对照组相比,aP < 0.05

    Figure  4.  Apoptosis of hepatocyte of rats in two groups at 3 d and 7 d after operation

    表  1  两组大鼠术后3 d和7 d血清中ALT、AST和ALB的含量

    Table  1.   Contents of ALT, AST and ALB in serum of rats in two groups at 3 d and 7 d after operation (x±s)

    组别 n ALT(mmol/L) AST(mmol/L) ALB(g/L)
    术后3 d 术后7 d 术后3 d 术后7 d 术后3 d 术后7 d
    nodosin组 12 136±8a 86±9a 167±8a 100±9a 19.7±1.9a 23.6±1.4a
    对照组 12 226±12 135±6 281±10 160±21 17.4±1.6 19.1±0.7
    注:与对照组比较,aP < 0.05
    下载: 导出CSV
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  • 收稿日期:  2016-01-15
  • 网络出版日期:  2021-01-19
  • 刊出日期:  2016-03-15

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