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甘草酸单铵对体外油酸钠诱导的小牛原代肝细胞脂肪沉积变性的保护作用。

Protective effects of monoammonium glycyrrhizinate on fatty deposit degeneration induced in primary calf hepatocytes by sodium oleate administration in vitro.

机构信息

Henan International Joint Laboratory of Animal Welfare and Health Breeding, Henan University of Science and Technology, Luoyang 471023, China.

Henan International Joint Laboratory of Animal Welfare and Health Breeding, Henan University of Science and Technology, Luoyang 471023, China.

出版信息

Res Vet Sci. 2022 Dec 5;150:213-223. doi: 10.1016/j.rvsc.2022.05.011. Epub 2022 Jul 3.

DOI:10.1016/j.rvsc.2022.05.011
PMID:35868152
Abstract

In this study we investigated the effect of MAG on fatty deposit-induced degeneration of primary calf hepatocytes induced by sodium oleate. Primary hepatocytes were isolated from dairy calves and cultured before allocation to the following treatment groups: control (untreated), model (starved for 12 h before treatment with 0.25 mM sodium oleate to induce steatosis-like changes, and the MAG group pretreated with MAG (0.1, 0.25, 0.5, and 1.5 mM) for 12 h before sodium oleate treatment (0.25 mM) for 12 h). To evaluate the effect of MAG on fat-induced degradation of primary hepatocytes, we evaluated lipid deposition, cell viability, apoptosis rate, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity in the culture supernatant, and expression of oxidant and antioxidant enzymes (LDH, MDA, GSH, and CAT), as well as the expression of lipid metabolism-related genes (PPARα, SREBP-1c, ChREBP, CPT1, CPT2, and MTP) and apoptosis-related genes (Cyt-c, Caspase 9, Caspase 8, Caspase 3, Bax, and Bcl-2). MAG significantly reduced lipid accumulation in hepatocytes induced by sodium oleate (P < 0.05), increased cell viability, decreased the apoptosis rate (P < 0.05), and significantly decreased ALT and AST activity in the culture supernatant (P < 0.05). MAG significantly decreased MDA levels in cells and LDH levels in the culture supernatant, while GSH and CAT levels were increased (P < 0.05). MAG significantly increased the expression of the lipid transport- and metabolism-related genes MTP, PPARα, CPT1 and CPT2, and decreased ChREBP expression (P < 0.05). At concentrations higher than 0.25 mM, MAG significantly decreased SREBP-1c expression (P < 0.05). MAG significantly decreased the expression of the apoptosis-related genes Cyt-c, Caspase 9, Caspase 8, Caspase3 and Bax, while Bcl-2 expression was increased (P < 0.05). These findings demonstrate that MAG improves the antioxidant capacity of hepatocytes and effectively reduces lipid deposition by inhibiting the expression of lipid metabolism- and apoptosis-related genes.

摘要

在这项研究中,我们研究了 MAG 对油酸钠诱导的初生小牛肝细胞脂肪沉积诱导的变性的影响。原代肝细胞从奶牛犊牛中分离出来,并在分配到以下治疗组之前进行培养:对照组(未处理)、模型组(在用 0.25mM 油酸钠处理前饥饿 12 小时,以诱导类似脂肪变性的变化,以及 MAG 组在用 0.25mM 油酸钠处理前用 MAG(0.1、0.25、0.5 和 1.5mM)预处理 12 小时)。为了评估 MAG 对脂肪诱导的原代肝细胞降解的影响,我们评估了脂质沉积、细胞活力、细胞凋亡率、培养液上清液中丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)活性,以及氧化应激和抗氧化酶(LDH、MDA、GSH 和 CAT)的表达,以及脂质代谢相关基因(PPARα、SREBP-1c、ChREBP、CPT1、CPT2 和 MTP)和凋亡相关基因(Cyt-c、Caspase 9、Caspase 8、Caspase 3、Bax 和 Bcl-2)的表达。MAG 显著减少了油酸钠诱导的肝细胞中的脂质堆积(P<0.05),增加了细胞活力,降低了细胞凋亡率(P<0.05),并显著降低了培养液上清液中的 ALT 和 AST 活性(P<0.05)。MAG 显著降低了细胞内 MDA 水平和培养液上清液中的 LDH 水平,同时增加了 GSH 和 CAT 水平(P<0.05)。MAG 显著增加了脂质转运和代谢相关基因 MTP、PPARα、CPT1 和 CPT2 的表达,并降低了 ChREBP 的表达(P<0.05)。在浓度高于 0.25mM 时,MAG 显著降低了 SREBP-1c 的表达(P<0.05)。MAG 显著降低了凋亡相关基因 Cyt-c、Caspase 9、Caspase 8、Caspase3 和 Bax 的表达,同时增加了 Bcl-2 的表达(P<0.05)。这些发现表明,MAG 通过抑制脂质代谢和凋亡相关基因的表达,提高了肝细胞的抗氧化能力,并有效减少了脂质沉积。

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