小分子多肽LK-5对CCl4致急性肝损伤小鼠的保护作用研究

doi:10.16431/j.cnki.1671-7236.2023.09.039

中国畜牧兽医 ›› 2023, Vol. 50 ›› Issue (9): 3833-3841.doi: 10.16431/j.cnki.1671-7236.2023.09.039

小分子多肽LK-5对CCl₄致急性肝损伤小鼠的保护作用研究

陈俊^1,4, 廖彭莹^2,3,5, 张新锐², 黄秀欢², 杨宇静², 刘华宇²

1. 广西中医药大学, 广西高校中药神经-代谢及免疫药理重点实验室, 南宁 530200;
2. 广西中医药大学药学院, 南宁 530200;
3. 广西高校中药提取纯化与质量分析重点实验室, 南宁 530200;
4. 广西中医药大学教学实验实训中心, 南宁 530200;
5. 中药学国家级实验教学示范中心(广西中医药大学), 南宁 530200

收稿日期:2023-02-16 出版日期:2023-09-05 发布日期:2023-08-24
通讯作者: 廖彭莹 E-mail:gxlpy@163.com
作者简介:陈俊,E-mail:543767193@qq.com。
基金资助:
国家自然科学基金项目(81960698);2022年自治区中药学研究生联合培养基地开放项目(桂学位[2021]6号);中药学广西一流学科建设项目(桂教科研[2022]1号);2017年广西中医药大学青年基金项目(2017QN011);广西高等学校千名中青年骨干教师培育计划项目(桂教师范[2019]81号);广西壮瑶药重点实验室科研任务项目(GXZYKF2022-10);广西中医药大学'桂派中医药传承创新团队’资助项目(2022A005);2023年度广西高校中青年教师科研基础能力提升项目(2023KY0289);广西高校中药药理重点实验室资助项目(05J16074)

Study on Protective Effects of Polypeptide LK-5 on Acute Liver Injury Induced by CCl₄ in Mice

CHEN Jun^1,4, LIAO Pengying^2,3,5, ZHANG Xinrui², HUANG Xiuhuan², YANG Yujing², LIU Huayu²

1. Key Laboratory of TCM Neuro-metabolism and Immunopharmacology of Guangxi Education Department, Guangxi University of Chinese Medicine, Nanning 530200, China;
2. College of Pharmacy, Guangxi University of Chinese Medicine, Nanning 530200, China;
3. Key Laboratory of TCM Extraction and Purification and Quality Analysis, Nanning 530200, China;
4. Teaching Experiment and Training Centre, Guangxi University of Chinese Medicine, Nanning 530200, China;
5. National Demonstration Center for Experimental Traditional Chinese Pharmacology(Guangxi University of Chinese Medicine), Nanning 530200, China

Received:2023-02-16 Online:2023-09-05 Published:2023-08-24

摘要/Abstract

摘要： 【目的】研究小分子多肽LK-5(氨基酸序列为LHMFK)对CCl₄致急性肝损伤模型小鼠的保护作用,探讨其作用机制。【方法】72只小鼠随机分为正常组、模型组、联苯双酯组(150 mg/kg)及LK-5低、中、高剂量组(30、60、120 mg/kg),每组12只。连续给药10 d,1 次/d,每次10 mL/kg。末次给药2 h后,各组(除正常组外)腹腔注射0.1% CCl₄花生油溶液,建立CCl₄致小鼠急性肝损伤模型,16 h后,按照试剂盒方法,检测小鼠血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、总胆汁酸(TBA)和碱性磷酸酶(AKP)水平,检测肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)、IL-10、血管紧张素1-7(Ang 1-7)和血管紧张素Ⅱ(Ang Ⅱ)水平。制作肝脏病理组织切片,苏木精-伊红染色(HE)后观察肝脏组织病理学变化。【结果】与正常组比较,模型组小鼠肝细胞有明显的脂肪变性,炎细胞分散分布于肝小叶血管周围和肝实质内,血清ALT、AST、AKP活性和TBA水平均显著上升,肝组织SOD、GSH-Px活性显著下降,MDA水平显著上升(P＜0.05),说明成功建立急性肝损伤模型,TNF-α、IL-6、Ang Ⅱ水平显著升高(P＜0.05)。与模型组比较,LK-5中、高剂量组能够显著降低血清ALT、AST活性(P＜0.05),各剂量组均可降低血清TBA水平和AKP活性(P＜0.05),LK-5高剂量组能够显著提高小鼠肝组织SOD、GSH-Px活性(P＜0.05),LK-5中、高剂量组能够显著降低肝组织MDA水平(P＜0.05),LK-5高剂量组TNF-α水平显著降低(P＜0.05),各剂量组能够显著降低肝组织IL-6、Ang Ⅱ水平(P＜0.05)。病理切片结果显示,LK-5各剂量组炎细胞均有不同程度减少,LK-5高剂量组肝小叶血管周围和肝实质内无炎细胞浸润。【结论】LK-5各剂量组对CCl₄致急性肝损伤小鼠具有保护作用,其中60、120 mg/kg LK-5效果更为明显,其作用机制可能与抗氧化、抗炎及对肾素-血管紧张素系统(RAS)的调控有关。

关键词: 小分子多肽LK-5; CCl₄; 急性肝损伤; 保护作用

Abstract: 【Objective】 To investigate the protective effect and potential mechanism of polypeptide LK-5 (the amino acids sequence:LHMFK) on acute liver injury induced by carbon tetrachloride (CCl₄) in mice.【Method】 72 mice were randomly divided into normal group,model group,biphenyl diester group (150 mg/kg),LK-5 low-dose,medium-dose and high-dose groups (30,60,120 mg/kg),with 12 mice in each group.Continuous administration of 10 mL/kg once a day for 10 days.Two hours after the last administration,each group (except for normal group) was intraperitoneally injected 0.1% CCl₄ peanut oil solution to establish CCl₄-induced acute liver injury model.16 h later,the serum levels of aspartate aminotransferase (AST),alanine aminotransferase (ALT),total bile acid (TBA),and alkaline phosphatase (AKP),the superoxide dismutase (SOD),glutathioneperoxidase (GSH-Px),malondialdehyde (MDA),tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),IL-10,angiotensin 1-7 (Ang 1-7), and angiotensin Ⅱ (Ang Ⅱ) in liver were measured with kit methods.The pathological sections of liver were made,and the histological changes of liver were observed after hematoxylin and eosin (HE) staining.【Result】 Compared with normal group,the hepatocytes in model group showed obvious steatosis,and inflammatory cells were scattered around the blood vessels of liver lobule and in liver parenchyma,the ALT,AST,AKP activities and TBA level in serum were significantly increased,the SOD and GSH-Px activities in liver were significantly decreased,the MDA level was significantly increased (P＜0.05),indicating the acute liver damage model was established successfully,and the levels of TNF-α,IL-6 and Ang Ⅱ were significantly increased (P＜0.05).Compared with model group,the ALT and AST activities in serum were significantly decreased in LK-5 medium- and high-dose groups (P＜0.05),the serum TBA level and AKP activity in each group were significantly decreased (P＜0.05),the SOD and GSH-Px activities in liver were significantly increased in LK-5 high-dose group (P＜0.05),the MDA level in liver was significantly decreased in LK-5 medium- and high-dose groups (P＜0.05),the TNF-α level in liver was significantly decreased in high-dose group (P＜0.05), and the levels of IL-6 and Ang Ⅱ were significantly decreased in each group (P＜0.05).The pathological section results showed that inflammatory cells in each dose group of LK-5 decreased,while in the high-dose group,there was no infiltration of inflammatory cells around the hepatic lobular blood vessels and in the liver parenchyma.【Conclusion】 LK-5 showed protective effect on acute liver injury induced by CCl₄ in mice in all dose groups,and 60, 120 mg/kg LK-5 had more significant effects. Its mechanism of action was related with the antioxidant,anti-inflammatory and regulation of renin-angiotensin system (RAS).

Key words: polypeptide LK-5; CCl₄; acute liver injury; protective effects

中图分类号:

S852
R965.1

陈俊, 廖彭莹, 张新锐, 黄秀欢, 杨宇静, 刘华宇. 小分子多肽LK-5对CCl₄致急性肝损伤小鼠的保护作用研究[J]. 中国畜牧兽医, 2023, 50(9): 3833-3841.

CHEN Jun, LIAO Pengying, ZHANG Xinrui, HUANG Xiuhuan, YANG Yujing, LIU Huayu. Study on Protective Effects of Polypeptide LK-5 on Acute Liver Injury Induced by CCl₄ in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3833-3841.

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小分子多肽LK-5对CCl₄致急性肝损伤小鼠的保护作用研究

Study on Protective Effects of Polypeptide LK-5 on Acute Liver Injury Induced by CCl₄ in Mice

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