基于ERK1/2信号通路探讨SHP2表达变化对人肝星状细胞LX-2活化的影响

肝脏 ›› 2025, Vol. 30 ›› Issue (12): 1674-1677.

基于ERK1/2信号通路探讨SHP2表达变化对人肝星状细胞LX-2活化的影响

郝礼森, 高莹莹, 展宗媛, 连俊, 刘甜, 王金妹, 羊东杰, 毛佳淇

063000 唐山华北理工大学附属医院消化内科(郝礼森,高莹莹,刘甜,王金妹,羊东杰,毛佳淇);310023 杭州杭州市西溪医院消化内科(展宗媛);725000 安康安康市中心医院超声医学科(连俊)

收稿日期:2025-01-30 发布日期:2026-02-10
基金资助:
河北省自然科学基金面上项目(H2018209366)

Effects of changes in SHP2 expression on the activation of human hepatic stellate cell LX-2 based on ERK1/2 signaling pathway

HAO Li-sen¹, GAO Ying-ying¹, ZHAN Zong-yuan², LIAN Jun³, LIU Tian¹, WANG Jin-mei¹, YANG Dong-jie¹, MAO Jia-qi¹

1. Department of Gastroenterology, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, China;
2. Department of Gastroenterology, Hangzhou Xixi Hospital, Hangzhou 310023, China;
3. Department of Ultrasound the Central, Hospital of Ankang City, Ankang 725000, China

Received:2025-01-30 Published:2026-02-10

摘要/Abstract

摘要： 目的探讨含SH2结构域的蛋白酪氨酸磷酸酶2(SHP2)表达变化对人肝星状细胞LX-2活化及细胞外信号调节激酶1/2(ERK1/2)的影响。方法将携带野生型SHP2基因的腺病毒Ad-SHP2及携带靶向SHP2的shRNA的腺病毒Ad-shRNA/SHP2分别转染人肝星状细胞LX-2;Western blot及实时荧光定量PCR检测LX-2细胞的SHP2及ERK1/2表达;免疫细胞化学染色及Western blot检测肝星状细胞活化标志物α-平滑肌肌动蛋白(α-SMA)表达;Western blot检测LX-2细胞的磷酸化ERK1/2(p-ERK1/2)表达。实验分组:①Control组:DMEM代替腺病毒转染LX-2细胞;②Ad-GFP组:空病毒Ad-GFP转染LX-2细胞;③Ad-shRNA/SHP2组:Ad-shRNA/SHP2转染LX-2细胞;④Ad-SHP2组:Ad-SHP2转染LX-2细胞。结果野生型SHP2及靶向SHP2的shRNA显著升高,LX-2细胞的SHP2表达降低(P<0.05)。免疫细胞化学染色检测显示,与control组及Ad-GFP组LX-2细胞α-SMA蛋白阳性表达IOD(0.073±0.003、0.074±0.004)比较,Ad-shRNA/SHP2组(0.048±0.003)显著降低(P<0.05),Ad-SHP2组(0.127±0.004)明显升高(P<0.05);Ad-shRNA/SHP2组LX-2细胞的α-SMA及p-ERK1蛋白表达量(0.154±0.033、0.134±0.023)明显低于control组(0.342±0.043、0.302±0.035)及Ad-GFP组(0.308±0.038、0.315±0.037,P<0.05),而Ad-SHP2组(0.517±0.036、0.522±0.053)显著高于control组及Ad-GFP组(P<0.05);四组LX-2细胞的ERK1 mRNA及蛋白表达差异无统计学意义(P>0.05)。结论 SHP2过表达促进LX-2细胞活化,而SHP2低表达则抑制其活化,ERK1/2信号通路介导了SHP2对LX-2细胞活化的调控。

关键词: 人肝星状细胞LX-2, SHP2, 肝星状细胞活化, 细胞外信号调节激酶1/2

Abstract: Objective To explore the effects of changes in protein tyrosine phosphatase 2 (SHP2) expression on activation and extracellular signal regulated kinase 1/2 (ERK1/2) in human hepatic stellate cell LX-2. Methods Adenovirus Ad-SHP2 with wild-type SHP2 gene and Ad-shRNA/SHP2 with shRNA targeting SHP2 were introduced into LX-2 cells cultured in vitro, respectively. Application of real-time fluorescence quantitative PCR and Western blot to analyze expression of SHP2 and ERK1/2 in LX-2 cells. Expression of phosphorylated ERK1/2 (p-ERK1/2) in LX-2 cells was analyzed by Western blot analysis. The expression of α-smooth muscle actin (α-SMA), an activated marker of hepatic stellate cells was detected immunocytochemical staining and Western blot. DMEM was used instead of adenovirus to transfect LX-2 cells in control group, the LX-2 cells was transfected with empty virus Ad-GFP in Ad-GFP group. Ad-SHP2 was transfected into LX-2 cells in Ad-shRNA/SHP2 group. Results Wild-type SHP2 and shRNA targeting SHP2 significantly increased and decreased the expression of SHP2 in LX-2 cells (P<0.05). Immunocytochemical staining showed that compared with the positive expression integral optical density (IOD) of α-SMA protein in LX-2 cells in control group and Ad-GFP group (0.073±0.003, 0.074±0.004), that in Ad-shRNA/SHP2 group (0.048±0.003) showed a significant (P<0.05) decrease and in Ad-SHP2 group (0.127±0.004) remarkably (P<0.05) increased. Expression levels of α-SMA and p-ERK1 protein in LX-2 cells of Ad-shRNA/SHP2 group (0.154±0.033, 0.134±0.023) significantly (P<0.05) lowered and in Ad-SHP2 group (0.517±0.036, 0.522±0.053) exhibited a significant (P<0.05) increase compared with those in control group (0.342±0.043, 0.302±0.035) and Ad-GFP group (0.308±0.038, 0.315±0.037). There was no significant difference in ERK1 mRNA and protein expression of LX-2 cells among 4 groups (P<0.05). Conclusion Overexpressed SHP2 can promote activation of LX-2 cells, while low expression of SHP2 can suppress activation of LX-2 cells, and the ERK1/2 signaling pathway mediates the effects of SHP2 on activation of LX-2 cells.

Key words: Human hepatic stellate cell LX-2, SHP2, Hepatic stellate cell activation, ERK1/2

郝礼森, 高莹莹, 展宗媛, 连俊, 刘甜, 王金妹, 羊东杰, 毛佳淇. 基于ERK1/2信号通路探讨SHP2表达变化对人肝星状细胞LX-2活化的影响[J]. 肝脏, 2025, 30(12): 1674-1677.

HAO Li-sen, GAO Ying-ying, ZHAN Zong-yuan, LIAN Jun, LIU Tian, WANG Jin-mei, YANG Dong-jie, MAO Jia-qi. Effects of changes in SHP2 expression on the activation of human hepatic stellate cell LX-2 based on ERK1/2 signaling pathway[J]. Chinese Hepatolgy, 2025, 30(12): 1674-1677.

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