重组人促血小板生成素对人肝细胞株L02增殖的影响

doi:10.3877/cma.j.issn.1674-6880.2020.03.002

中华危重症医学杂志(电子版) ›› 2020, Vol. 13 ›› Issue (03) : 165 -169. doi: 10.3877/cma.j.issn.1674-6880.2020.03.002

所属专题：文献；

论著

重组人促血小板生成素对人肝细胞株L02增殖的影响

俞丹丽¹, 蔡玲², 戚东升¹, 郑树森³, 章渭方¹^,^†(

)

^1. 310003　杭州，浙江大学医学院附属第一医院重症医学科
^2. 310007　杭州，杭州市中医院重症医学科
^3. 310003　杭州，浙江大学医学院附属第一医院肝胆胰外科

收稿日期:2019-12-25 出版日期:2020-06-01
通信作者: 章渭方
基金资助:
浙江省自然科学基金项目(LY15H15001)

Effect of recombinant human thrombopoietin on proliferation in human hepatic cell line L02

Danli Yu¹, Ling Cai², Dongsheng Qi¹, Shusen Zheng³, Weifang Zhang¹^,^†()

^1. Department of Critical Care Medicine, the First Affiliated Hospital, College of Medicine, Zhejiang University, 310003 Hangzhou, China
^2. Department of Critical Care Medicine, Hangzhou Hospital of Traditional Chinese Medicine, 310007 Hangzhou, China
^3. Department of Hepatopancreatobiliary Surgery, the First Affiliated Hospital, College of Medicine, Zhejiang University, 310003 Hangzhou, China

Received:2019-12-25 Published:2020-06-01
Corresponding author: Weifang Zhang
About author:
Corresponding author: Zhang Weifang, Email: zhangweifang@zju.edu.cn

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俞丹丽, 蔡玲, 戚东升, 郑树森, 章渭方. 重组人促血小板生成素对人肝细胞株L02增殖的影响[J]. 中华危重症医学杂志(电子版), 2020, 13(03): 165-169.

Danli Yu, Ling Cai, Dongsheng Qi, Shusen Zheng, Weifang Zhang. Effect of recombinant human thrombopoietin on proliferation in human hepatic cell line L02[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2020, 13(03): 165-169.

目的

探究重组人促血小板生成素（rhTPO）对健康人肝细胞株L02的影响。

方法

以健康人肝细胞株L02为研究对象，通过不同剂量rhTPO[对照组（0 U rhTPO）、低剂量组（150 U rhTPO）及高剂量组（750 U rhTPO）]处理L02细胞，观察各组L02细胞的存活率。采用Western-blotting检测各组细胞[A组（0 U rhTPO）、B组（15 U rhTPO）、C组（150 U rhTPO）及D组（1 500 U rhTPO）]的细胞外信号调节激酶（ERK）及其磷酸化产物（p-ERK）蛋白的表达水平，并采用荧光定量PCR检测ERK信使RNA（mRNA）表达水平。

结果

各组L02细胞存活率的比较，差异有统计学意义（F = 8.843，P = 0.016）。进一步两两比较发现，低剂量组及高剂量组的存活率均显著高于对照组[（107.9 ± 4.0）%、（112.0± 1.6）%、（100.0 ± 2.5）%]，且高剂量组细胞的存活率更高（P均< 0.05）。各组L02细胞间ERK蛋白及ERK mRNA表达水平的比较，差异均无统计学意义（F = 1.140，P = 0.392；F = 0.437，P = 0.733），而各组L02细胞间p-ERK蛋白表达水平的比较差异有统计学意义（F = 31.400，P < 0.001），且B组、C组及D组细胞的p-ERK蛋白表达水平均显著高于A组细胞（P均< 0.05），而B组、C组及D组细胞p-ERK蛋白表达水平两两比较，差异均无统计学意义（P均> 0.05）。

结论

rhTPO可通过激活ERK促进L02细胞数量增殖。

关键词: 人肝细胞株, 重组人促血小板生成素, 细胞增殖

Objective

To investigate the effect of recombinant human thrombopoietin (rhTPO) on healthy human hepatic cell line L02.

Methods

The survival rate of L02 cells was observed among a control group (0 U rhTPO), a low-dose group (150 U rhTPO) and a high-dose group (750 U rhTPO) according to different doses of rhTPO. The expression levels of extracellular signal regulated kinase (ERK) and phosphorylation-ERK (p-ERK) protein were detected by Western-blotting, and the expression level of ERK messenger RNA (mRNA) was examined by fluorogenic quantitative PCR in the L02 cells among group A (treated with 0 U rhTPO), group B (treated with 15 U rhTPO), group C (treated with 150 U rhTPO) and group D (treated with 1 500 U rhTPO).

Results

The survival rate of L02 cells was significantly different among the three groups (F = 8.843, P = 0.016). It was much higher in the low-dose group and high-dose group than in the control group, and highest in the high-dose group [(107.9 ± 4.0)%, (112.0 ± 1.6)%, (100.0 ± 2.5)%; all P < 0.05]. However, there were no statistically significant differences in the expression levels of ERK protein and ERK mRNA among the four groups (F = 1.140, P = 0.392; F = 0.437, P = 0.733). Besides, the expression level of p-ERK protein was significantly different among the four groups (F = 31.400, P < 0.001). It increased obviously in group B, group C and group D as compared with group A (all P < 0.05), while no significant differences were noted among group B, group C and group D (all P > 0.05).

Conclusion

rhTPO can promote the proliferation of L02 hepatocytes by activating ERK.

Key words: Human hepatic cell line, Recombinant human thrombopoietin, Cell proliferation

表1 不同浓度rhTPO刺激下各组L02细胞存活率的比较（%，±s）

组别	细胞数（个/ mL）	存活率
对照组	1 × 10⁴	100.0 ± 2.5
低剂量组	1 × 10⁴	107.9 ± 4.0^a
高剂量组	1 × 10⁴	112.0 ± 1.6^ab
F值	?	8.843
P值	?	0.016

表1 不同浓度rhTPO刺激下各组L02细胞存活率的比较（%，±s）

组别	细胞数（个/ mL）	存活率
对照组	1 × 10⁴	100.0 ± 2.5
低剂量组	1 × 10⁴	107.9 ± 4.0^a
高剂量组	1 × 10⁴	112.0 ± 1.6^ab
F值	?	8.843
P值	?	0.016

图1 各组L02细胞ERK及p-ERK蛋白表达水平的比较

图2 各组L02细胞ERK mRNA表达水平的比较

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