E型前列腺素受体4对CCL4诱导的肝损伤小鼠肝组织肝纤维化相关蛋白表达的影响*

doi:10.3969/j.issn.1672-5069.2023.01.005

实用肝脏病杂志 ›› 2023, Vol. 26 ›› Issue (1): 15-18.doi: 10.3969/j.issn.1672-5069.2023.01.005

E型前列腺素受体4对CCL₄诱导的肝损伤小鼠肝组织肝纤维化相关蛋白表达的影响^*

李蕊, 秦秋实, 张玥, 曹颖, 闫杰, 朱鏐娈

100015 北京市首都医科大学附属北京地坛医院传染病研究所/新发突发传染病重点实验室/北京市感染性疾病研究中心(李蕊,张玥,朱鏐娈);肝病中心(闫杰);北京大学地坛医院教学医院(秦秋实); 清华大学附属北京清华长庚医院肝胆内科(曹颖)

收稿日期:2022-07-28 出版日期:2023-01-10 发布日期:2023-02-07
通讯作者: 朱鏐娈,E-mail: zhuliuluan@ccmu.edu.cn
作者简介:李蕊,女,38岁,医学硕士,助理研究员。E-mail: lirui1418@163.com
基金资助:
^*北京市优秀人才培养基金资助项目(编号:2018000021223ZK27)

Role of E series of prostaglandin receptor 4 on liver fibrosis-related protein expression in mice with carbon tetrachloride-induced liver injuries

Li Rui, Qin Qiushi, Zhang Yue, et al

Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Ditan Hospital, Capital Medical University, Beijing 100015, China

Received:2022-07-28 Online:2023-01-10 Published:2023-02-07

摘要/Abstract

摘要： 目的探讨E型前列腺素受体4(EP4)在CCL₄诱导的肝损伤小鼠肝纤维化发生过程中的作用。方法取30只C57BL/6N小鼠,随机分为对照组、模型组和CCL₄联合E7046处理组(n=10),采用CCL₄注射法建立肝损伤模型,分别给予甲基纤维素或EP4特异性拮抗剂E7046溶液灌胃干预。采用Western blot和qPCR检测小鼠肝组织EP4、α-SMA蛋白及其mRNA水平。常规行HE染色、Masson染色和天狼星红染色,鉴定小鼠肝组织病理学变化。结果模型组小鼠肝组织EP4蛋白表达比对照组显著增高,经E7046干预后,肝组织EP4蛋白表达降低,同样地,模型组小鼠肝组织EP4编码基因Ptger 4 mRNA水平为(3.5±0.1),显著高于对照组【(1.0±0.1),P<0.05】,而CCL₄联合E7046处理组为(2.4±0.2),较模型组显著降低(P<0.05);模型组小鼠血清ALT和AST水平分别为(1753.5±328.6)U/L和(1586.2±204.1)U/L,显著高于对照组(P<0.05),而在CCL₄联合E7046处理组,两种酶水平较模型组显著降低【分别为(885.9±269.6)U/L和(892.4±208.6)U/L,P<0.05】;模型组小鼠肝组织α-SMA表达较对照组增高,而在CCL₄联合E7046组α-SMA水平较模型组降低,模型组小鼠肝组织Acta2和Col1a1 mRNA 水平较对照组显著上调,而CCL₄联合E7046组则较模型组显著降低(P<0.05)。结论 EP4在CCL₄诱导的小鼠肝纤维化发生过程中发挥重要作用,阻断EP4与其配体结合有助于改善肝纤维化。

关键词: 肝纤维化, 四氯化碳, E型前列腺素受体4, Ptger 4基因, 小鼠

Abstract: Objective This experiment aimed at exploring the role of E series of prostaglandin receptor 4 (EP4) on liver fibrosis-related protein expression in mice with CCL₄-induced liver injuries. Methods 30 C57BL/6N mice were randomly divided into control, CCL₄-intervened, and CCL₄ and E7046 conbination-intervened group, with 10 mice in each group. The liver injury model was established by CCL₄ intraperitoneal injection, with methylcellulose or EP4-specific antagonist, E7046 solution gavage. After 27 day experiment, the mice were sacrificed under anesthesia, and sera and liver tissues were obtained. The EP4 and α-SMA protein expression and their mRNA levels were detected by Western blot and real-time PCR. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical instrument. The liver histopathological examination was performed by HE, Masson and Sirius red staining. Results The hepatic expression of EP4 protein in model mice increased greatly compared to in the control, while it decreased obviously in the combination-intervened mice compared to in the model; similarly, the EP4 encoded Ptger 4 mRNA level in the model group was (3.5±0.1), significantly higher than [(1.0±0.1), P<0.05] in the control, while that was (2.4±0.2) in the combination group, significantly decreased compared to in the model(P<0.05); serum ALT and AST levels in the model were (1753.5±328.6)U/L and (1586.2±204.1)U/L, much higher than in the control(P<0.05), while they decreased greatly in CCL₄ and E7046 combination group, e.g., [(885.9±269.6)U/L and (892.4±208.6)U/L, respectively, P<0.05]; the hepatic α-SMA expression in the model was stronger than in the control, while it became obviously weaker in CCL₄ and E7046 combination group, and the hepatic Acta2 and Col1a1 mRNA levels in the model up-regulated greatly compared to in the control, and also they became decreased in the CCL₄ and E7046-intervened group (P<0.05). Conclusion EP4 might play an pivotal role in CCL₄-induced liver fibrosis, which seem to be a target for ameliorating liver fibrosis.

Key words: Liver fibrosis, Carbon tetrachloride, E-type prostaglandin receptor 4, Ptger 4 gene, Mice

李蕊, 秦秋实, 张玥, 曹颖, 闫杰, 朱鏐娈. E型前列腺素受体4对CCL₄诱导的肝损伤小鼠肝组织肝纤维化相关蛋白表达的影响^*[J]. 实用肝脏病杂志, 2023, 26(1): 15-18.

Li Rui, Qin Qiushi, Zhang Yue, et al. Role of E series of prostaglandin receptor 4 on liver fibrosis-related protein expression in mice with carbon tetrachloride-induced liver injuries[J]. Journal of Practical Hepatology, 2023, 26(1): 15-18.

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E型前列腺素受体4对CCL₄诱导的肝损伤小鼠肝组织肝纤维化相关蛋白表达的影响^*

Role of E series of prostaglandin receptor 4 on liver fibrosis-related protein expression in mice with carbon tetrachloride-induced liver injuries

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