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[粒细胞集落刺激因子动员的自体骨髓干细胞促进部分肝移植大鼠的肝脏再生:一项实验研究]

[Granulocyte colony-stimulating factor-mobilized autologous bone marrow stem cells promote the liver regeneration of partial liver transplant: an experiment with rats].

作者信息

Liu Feng, Pan Xiao-ben, Chen Guo-dong, Jiang Dong, Cong Xu, Fei Ran, Wei Lai

机构信息

Institute of Hepatology, People's Hospital, Peking University, Beijing 100044, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2005 Dec 14;85(47):3342-5.

Abstract

OBJECTIVE

To investigate the effects of granulocyte colony-stimulating factor (G-CSF)-mobilized autologous bone marrow stem cells on the liver regeneration of partial liver transplant.

METHODS

A 50% partial liver transplantation model as established by transplanting parts of the liver of female rats to the male rats with the equal weights that had had parts of their livers resected. Then the transplanted male rats were randomly divided into 3 equal groups: G-CSF + PLTx group that was injected hypodermically with recombinant human G-CSF (rhG-CSF) daily for 5 days and then underwent liver resection and 50% partial liver transplantation (PLTx); PLTx + G-CSF group that underwent PLTx and 3 hours after the transplantation received injection of rhG-CSF daily for 5 days, and PLTx control group that underwent PLTx and 3 hours later received injection of normal saline daily for 5 days. One, 3, 5, 7, and 14 days after the PLTx blood samples and livers were collected from 6 rats from each group, and one hour before the liver was taken bromodeoxyuridine (BrdU) was injected intraperitoneally to be integrated into the synthesis of DNA in the liver cells. Immunohistochemistry was used to detect the CD34 and BrdU-positive cells. In situ hybridization was used to detect the sry (sex-determining region) gene so as to determine the origin of the proliferating cells in the transplanted liver.

RESULTS

The 14-day survival rate of the G-CSF + PLTx group was 90%, significantly higher than those of the G-CSF + PLTx (60%) and PLTx group (50%) (both P < 0.05) with a significant difference between the latter 2 groups too (P < 0.05). The mitosis index of liver cells 3 days after the transplantation of the PLTx + G-CSF group was 30% +/- 5%, significantly higher than those of the G-CSF + PLTx group (24% +/- 7%) and PLTx group (24% +/- 6%) (both P < 0.05) without a significant difference between the latter 2 groups. The rate of BrdU-positive cells of the PLTx + G-CSF group was 42% +/- 6%, significantly higher than those of the G-CSF + PLTx group (38% +/- 4%) and PLTx group (34% +/- 8%) (both P < 0.05). The transplantation the mitosis index and rate of BrdU-positive cells decreased since the 7th days after transplantation in all 3 groups, however, with the same relationship among them. The numbers of CD34(+) cell around the portal area of the 2 G-CSF groups increased since the 3rd day after transplantation in comparison with the PLTx group, was the highest on the 3rd day for the G-CSF + PLTx group, and continued to increase in the PLTx + G-CSF group. Since the 3rd day after transplantation, sry-positive cells were seen in the hepatic sinusoid and portal area in the 2 G-CSF groups, and were rarely seen in the PLTx group.

CONCLUSION

G-CSF treatment after 50% PLTx significantly promotes liver regeneration and ameliorates the liver damage, thus raising the survival rate.

摘要

目的

探讨粒细胞集落刺激因子(G-CSF)动员的自体骨髓干细胞对部分肝移植后肝脏再生的影响。

方法

通过将雌性大鼠部分肝脏移植到体重相等且部分肝脏已切除的雄性大鼠体内,建立50%部分肝移植模型。然后将移植后的雄性大鼠随机分为3组,每组数量相等:G-CSF + PLTx组,每天皮下注射重组人G-CSF(rhG-CSF)共5天,然后进行肝切除和50%部分肝移植(PLTx);PLTx + G-CSF组,先进行PLTx,移植后3小时开始每天注射rhG-CSF共5天;PLTx对照组,进行PLTx,3小时后每天注射生理盐水共5天。在PLTx后1、3、5、7和14天,从每组6只大鼠采集血液样本和肝脏,在取肝脏前1小时腹腔注射溴脱氧尿苷(BrdU),使其掺入肝细胞DNA合成中。采用免疫组织化学检测CD34和BrdU阳性细胞。采用原位杂交检测sry(性别决定区)基因,以确定移植肝脏中增殖细胞的来源。

结果

G-CSF + PLTx组14天生存率为90%,显著高于PLTx + G-CSF组(60%)和PLTx组(50%)(均P < 0.05),后两组之间也有显著差异(P < 0.05)。PLTx + G-CSF组移植后3天肝细胞有丝分裂指数为30%±5%,显著高于G-CSF + PLTx组(24%±7%)和PLTx组(24%±6%)(均P < 0.05),后两组之间无显著差异。PLTx + G-CSF组BrdU阳性细胞率为42%±6%,显著高于G-CSF + PLTx组(38%±4%)和PLTx组(34%±8%)(均P < 0.05)。3组移植后7天起有丝分裂指数和BrdU阳性细胞率均下降,但它们之间的关系相同。与PLTx组相比,2个G-CSF组门静脉周围CD34(+)细胞数量从移植后第3天开始增加,G-CSF + PLTx组第3天最高,PLTx + G-CSF组持续增加。移植后第3天起,2个G-CSF组肝窦和门静脉区可见sry阳性细胞,PLTx组少见。

结论

50% PLTx后给予G-CSF治疗可显著促进肝脏再生,减轻肝损伤,从而提高生存率。

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