四氯化碳诱导的大鼠肝纤维化肝组织中SHP2表达与在体肝星状细胞活化及增殖的关系

肝脏 ›› 2023, Vol. 28 ›› Issue (5): 549-553.

四氯化碳诱导的大鼠肝纤维化肝组织中SHP2表达与在体肝星状细胞活化及增殖的关系

张朋垒, 张明婷, 郝礼森, 靳丽敏, 潘恩亮, 何宇, 苗笑佳, 王薇

063000 唐山华北理工大学附属医院消化内科

收稿日期:2022-05-06 出版日期:2023-05-31 发布日期:2023-08-29
通讯作者: 郝礼森,Email:haolisen125@163.com
基金资助:
河北省自然科学基金面上项目(H2018209366)

Relationship between SHP2 expression and the activation and proliferation of hepatic stellate cells in liver tissues of rats with CCl₄-induced liver fibrosis

ZHANG Peng-lei, ZHANG Ming-ting, HAO Li-sen, JIN Li-min, PAN En-liang, HE Yu, MIAO Xiao-jia, WANG Wei

Department of Gastroenterology, the Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, China

Received:2022-05-06 Online:2023-05-31 Published:2023-08-29
Contact: HAO Li-sen,Email:haolisen125@163.com

摘要/Abstract

摘要： 目的探讨四氯化碳诱导的大鼠肝纤维化肝组织及在体肝星状细胞(HSC)的含SH2结构域的蛋白酪氨酸磷酸酶2(SHP2)表达变化与在体HSC活化及增殖的关系。方法随机将50只健康雄性SD大鼠分为对照组(10只)、模型组(40只),采用腹腔注射四氯化碳法构建大鼠肝纤维化模型,Masson三色及HE染色检测大鼠肝组织的病理组织学变化,免疫组织化学染色检测大鼠肝组织的α-平滑肌肌动蛋白(α-SMA)及SHP2表达,SHP2与α-SMA免疫荧光双标记检测大鼠肝组织中活化HSC的SHP2表达。结果与对照组大鼠肝组织的α-SMA阳性表达积分光密度值 (IOD) (0.09±0.01)相比,造模不同时间(2周、4周、6周、8周)大鼠纤维化肝组织的α-SMA阳性表达IOD (0.13±0.01、0.18±0.01、0.24±0.02、0.28±0.02)显著增加(P<0.05),并随着造模时间延长逐渐升高(P<0.05) ,即在体HSC的活化及增殖逐渐加快(α-SMA是HSC的活化标志)。造模不同时间(2周、4周、6周、8周)大鼠纤维化肝组织的SHP2阳性表达IOD (0.23±0.01、0.27±0.01、0.30±0.01、0.33±0.01)较对照组(0.19±0.01)显著增加(P<0.05),且逐渐升高(P<0.05)。SHP2与α-SMA免疫荧光双标记检测显示,造模不同时间(2周、4周、6周、8周)大鼠纤维化肝组织中表达SHP2的活化HSC占总的活化HSC的百分比(54%±3%、 62%±2%、73%±4%、 86%±3%)逐渐升高(P<0.05)。结论在四氯化碳诱导的大鼠肝纤维化模型中,肝组织及在体HSC 的SHP2表达与在体HSC的活化及增殖呈显著正相关。

关键词: 肝纤维化, 肝星状细胞, 含SH2结构域的蛋白酪氨酸磷酸酶2, 细胞增殖

Abstract: Objective To explore the relationship between SH2 domain-containing protein tyrosine phosphatase 2 (SHP2)expression and the activation and proliferation of hepatic stellate cells (HSC)in liver tissues.Methods A liver fibrosis rat model was established by using CCl₄ treatment. 50 healthy male SD rats were randomly divided into control group (10)and model group (40). Masson's trichrome staining and Hematoxylin and eosin (HE)staining were used to detect histological changes in liver tissues of rats. The expressions of SHP2 and alpha-smooth muscle action (α-SMA)in hepatic tissues of rats were analyzed through immunohistochemical staining. The expressions of SHP2 in HSC in vivo were analyzed through α-SMA and SHP2 immunofluorescence double-labeling.Results Compared with the integrated optical density (IOD)of α-SMA in the liver tissues of the control group (0.09±0.01), the IOD of α-SMA in the fibrotic liver tissues of rats at weekly time points (2, 4, 6, 8 weeks)during making model (0.13±0.01, 0.18±0.01, 0.24±0.02, 0.28±0.02)notably increased (P<0.05), and the more severe liver fibrosis, the higher α-SMA expression of hepatic tissues in rats (P<0.05). Since the α-SMA is a marker of activated HSC, the activation and proliferation of HSC in vivo gradually accelerated with the progress of liver fibrosis. Compared with the IOD of SHP2 in the liver tissues of the control group (0.19±0.01), the IOD of SHP2 in the fibrotic liver tissues of rats in the model group (0.23±0.01, 0.27±0.01, 0.30±0.01, 0.33±0.01)significantly increased (P<0.05), and the more severe liver fibrosis, the higher SHP2 expression in hepatic tissues of rats (P<0.05). The α-SMA and SHP2 immunofluorescence double-labeling showed that, at weekly time points (2, 4, 6, 8 weeks)during model formation, the percentage of activated HSC expressing SHP2 to the all activated HSC (54%±3%, 62%±2%, 73%±4%, 86%±3%)gradually heightened (P<0.05).Conclusion In rats with CCl₄-induced liver fibrosis, the expressions of SHP2 in both liver tissues and HSC in vivo have an evident positive correlation with the activation and proliferation of HSC.

Key words: Liver fibrosis, Hepatic stellate cells, SHP2, Proliferation

张朋垒, 张明婷, 郝礼森, 靳丽敏, 潘恩亮, 何宇, 苗笑佳, 王薇. 四氯化碳诱导的大鼠肝纤维化肝组织中SHP2表达与在体肝星状细胞活化及增殖的关系[J]. 肝脏, 2023, 28(5): 549-553.

ZHANG Peng-lei, ZHANG Ming-ting, HAO Li-sen, JIN Li-min, PAN En-liang, HE Yu, MIAO Xiao-jia, WANG Wei. Relationship between SHP2 expression and the activation and proliferation of hepatic stellate cells in liver tissues of rats with CCl₄-induced liver fibrosis[J]. Chinese Hepatolgy, 2023, 28(5): 549-553.

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