珍菊汤对小鼠急性肝损伤保护作用研究

doi:10.16431/j.cnki.1671-7236.2023.11.039

摘要/Abstract

摘要： 【目的】探讨自组方剂珍菊汤对3种不同诱因导致的急性肝损伤模型小鼠的保护作用。【方法】利用高效液相色谱（HPLC）测定珍菊汤主要成分及其含量。采用1，1-二苯基-2-三硝基苯肼（DPPH）清除率、羟基自由基清除率、总还原能力测定和对5-脂氧合酶（5-LOX）活性的抑制率探究珍菊汤的抗氧化和抗炎作用。选取42只小鼠随机分为7组进行24 h经口急性毒性试验及21 d蓄积毒性试验对其安全性进行评价。另选取113只小鼠，其中随机选取8只为空白对照组（Control），其余105只随机平均分配至3种药物诱导的急性肝损伤试验，每个试验35只。各急性肝损伤试验内再随机分为模型组（Model）、阳性药物水飞蓟宾组（Silybin）及珍菊汤高（H）、中（M）和低（L）剂量组，每组7只。Control和Model组连续灌胃生理盐水14 d；阳性药物Silybin组按50 mg/kg剂量连续灌胃14 d；珍菊汤高、中、低剂量组分别按照生药浓度10.0、5.0和2.5 g/kg连续灌胃珍菊汤14 d；第15天分别以四氯化碳（CCl₄）、对乙酰氨基酚（APAP）和酒精（Alcohol）诱导急性肝损伤模型，于第16或20天剖解，检测肝脏病理，血清生化指标、抗氧化指标、炎性因子及肝脏特异性基因miRNA（miR-122）变化。【结果】高效液相色谱测定得出每1 g珍菊汤组方中含有芦丁0.27 mg，木犀草苷2.72 mg，异绿原酸A 0.26 mg，盐酸小檗碱0.28 mg。珍菊汤DPPH自由基清除半数抑制浓度（IC₅₀）和羟自由基清除IC₅₀分别为2.394和104 mg/mL，总还原能力为3.73±0.04，抗炎率为75.97%±3.11%。经口急性毒性试验和蓄积毒性试验表明珍菊汤安全无毒。CCl₄、APAP和Alcohol所致急性肝损伤病理模型中，Model组小鼠肝组织出现不同程度病变，不同剂量珍菊汤组和阳性药物Silybin组均有一定程度改善。与Model组相比，珍菊汤高剂量干预后，肝脏血清生化指标谷草转氨酶（AST）、谷丙转氨酶（ALT）、甘油三酯（TG）和总胆固醇（T-CHO）显著降低（P<0.05）；抗氧化指标丙二醛（MDA）显著降低（P<0.05），超氧化物歧化酶（SOD）、总抗氧化能力（T-AOC）和谷胱甘肽过氧化物酶（GSH-Px）显著升高（P<0.05）；炎性因子白细胞介素-1β（IL-1β）、IL-6和肿瘤坏死因子-α（TNF-α）和肝脏miR-122基因显著降低（P<0.05）。【结论】珍菊汤是一种安全无毒的组方药物，灌胃10.0 g/kg能够显著改善小鼠肝脏病变，提高小鼠肝脏抗氧化能力，对CCl₄、APAP和Alcohol所致急性肝损伤具有较好的保护作用。

关键词: 珍菊汤; 急性肝损伤; 抗氧化; 抗炎; miR-122

Abstract: 【Objective】 The aim of this study was to explore the protective effect of self-constituted prescription Zhen-Ju decoction on mice with acute liver injury caused by three different causative factors.【Method】 The main components and contents of Zhen-Ju decoction were determined by high-performance liquid chromatography (HPLC).The antioxidant and anti-inflammatory effects of Zhen-Ju decoction were investigated using 1, 1-diphenyl-2-picrylhydrazyl radical(DPPH) scavenging rate, hydroxyl radical scavenging rate, total reducing capacity and inhibition of 5-lipoxygenase(5-LOX) activity.42 mice were randomly divided into 7 groups to perform 24 h oral acute toxicity test and 21 d accumulation toxicity test to evaluate its safety.An additional 113 mice were selected, of which 8 were randomly selected as the blank control group (Control), and the remaining 105 were randomly assigned equally to three drug-induced acute liver injury trials, with 35 mice in each trial.Each acute liver injury test group was randomly divided into Model group, positive drug Silybum marianum group (Silybin) and high, medium and low dose Zhen-Ju decoction groups, with 7 rats in each group.Control and Model groups were oral administrated with saline for 14 d, Silybin group was oral administrated with 50 mg/kg Silybin for 14 d, the high, medium, and low dose groups were oral administrated with 10.0, 5.0, and 2.5 g/kg of Zhen-Ju decoction for 14 d, respectively. On the 15th day, acute liver injury models were induced with carbon tetrachloride (CCl₄), acetaminophen (APAP), and alcohol (Alcohol), respectively. Mice were executed at 16 or 20 d, then detected changes in liver pathology, serum biochemical parameters, antioxidant parameters, inflammatory factors and liver-specific gene miRNA (miR-122).【Result】 According to the determination by HPLC, each 1 g of Zhen-Ju decoction formula contained 0.27 mg of rutin, 2.72 mg of lignan, 0.26 mg of isochlorogenic acid A and 0.28 mg of berberine hydrochloride.The DPPH radical scavenging half maximal inhibitory concentration(IC₅₀) and hydroxyl radical scavenging IC₅₀ of Zhen-Ju decoction were 2.394 and 104 mg/mL, respectively, with a total reducing capacity of 3.73±0.04 and an anti-inflammatory rate of 75.97%±3.11%.The oral acute toxicity test and accumulation toxicity test showed that Zhen-Ju decoction was safe and non-toxic.In the pathological model of acute liver injury caused by CCl₄, APAP and Alcohol, the liver tissues of mice in Model group showed different degrees of lesions, and different doses of Zhen-Ju decoction and positive drug Silybin group showed some degree of improvement.Compared with Model group, after the high-dose intervention of Zhen-Ju decoction, the liver serum biochemical indicators glutamic oxalyl transaminase (AST), glutamic alanine transaminase (ALT), triglyceride (TG) and total cholesterol (T-CHO) were significantly lower (P<0.05), the antioxidant index malondialdehyde (MDA) was significantly lower (P<0.05) and superoxide dismutase (SOD), total antioxidant capacity (T-AOC) and glutathione peroxidase (GSH-Px) were significantly higher (P<0.05), the inflammatory factors interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) and miR-122 were significantly lower (P<0.05).【Conclusion】 Zhen-Ju decoction was a safe and non-toxic formula, and oral administrated with 10.0 g/kg Zhen-Ju decoction could significantly improve liver lesions and increase the antioxidant capacity of mouse liver, and had a good protective effect against acute liver injury caused by CCl₄, APAP and Alcohol.

Key words: Zhen-Ju decoction; acute liver injury; antioxidant; anti-inflammatory; miR-122

中图分类号:

S859.7

夏招彬, 孙艳, 韩其君, 汪露, 赵星, 仁科, 唐木克, 索郎扎西, 陈朝喜. 珍菊汤对小鼠急性肝损伤保护作用研究[J]. 中国畜牧兽医, 2023, 50(11): 4724-4736.

XIA Zhaobin, SUN Yan, HAN Qijun, WANG Lu, ZHAO Xing, REN Ke, TANG Muke, SUO Langzhaxi, CHEN Chaoxi. Study on the Protective Effect of Zhen-Ju Decotion on Acute Liver Injury in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4724-4736.

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