组蛋白乙酰化酶抑制剂(DCH36_06)对小鼠急性肝损伤的保护作用及机制研究

肝脏 ›› 2019, Vol. 24 ›› Issue (10): 1125-1128.

组蛋白乙酰化酶抑制剂(DCH36_06)对小鼠急性肝损伤的保护作用及机制研究

彭金金, 黄鹤鸣, 刘彦君, 石翠翠, 范建高, 张元元, 罗成, 李光明

200092 上海交通大学医学院附属新华医院消化内科(彭金金,黄鹤鸣,刘彦君,石翠翠,范建高,李光明);中国科学院上海药物研究所药物研发与设计中心(张元元,罗成)

收稿日期:2019-07-08 发布日期:2020-03-27
通讯作者: 李光明,Email:liguangming@xinhuamed.com.cn

Protective effect and mechanism of histone acetyltransferase inhibitor DCH36_06 on mice with acute liver injury

PENG Jin-jin¹, HUANG He-ming¹, LIU Yan-jun¹, SHI Cui-cui¹, FAN Jian-gao¹, ZHANG Yuan-yuan², LUO Cheng², LI Guang-ming¹

1. Department of Gastroenterology, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, 1665 Kongjiang Road, Shanghai 200092, China;
2. Drug Discovery and Design Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China

Received:2019-07-08 Published:2020-03-27
Contact: LI Guang-ming, Email: liguangming@xinhuamed.com.cn

摘要/Abstract

摘要： 目的探讨P300/CBP组蛋白乙酰化酶抑制剂DCH36_06对LPS/D-Gal诱导的小鼠急性肝损伤的保护作用及其机制。方法 75只C57BL/6小鼠随机分成4组,正常对照组20只、ALI模型组20只、药物干预组20只和药物对照组15只,分别予腹腔注射0.9%氯化钠溶液、LPS/D-Gal、LPS/D-Gal+DCH36_06和DCH36_06处理。前3组小鼠于LPS/D-Gal刺激4 h后每组各处死5只,采集小鼠血清和肝组织,检测肝功能指标并进行HE、TUNEL染色评估肝组织病理学变化和肝细胞凋亡。RT-PCR和ELISA检测肝组织中促炎细胞因子的表达水平。余小鼠继续饲养至24 h,以观察各组小鼠24 h生存率。结果急性肝损伤模型组15只小鼠24 h存活6只,DCH36_06药物干预组15只小鼠24 h存活13只。LPS/D-Gal诱导4 h后模型组肝组织中可见显著肝细胞变性坏死,炎症细胞浸润并见散在肝细胞凋亡,与模型组相比,干预组小鼠肝组织坏死性炎症及凋亡均明显减轻。干预组小鼠血清AST、ALT和TBil水平分别为(281.4±48.1)U/L、(175.2±32.5)U/L和(9.8±0.7)μmol/L,模型组小鼠分别为(1151.0±111.1)U/L、(921.5±47.9)U/L和(21.0±0.4)μmol/L差异有统计学意义(P<0.05)。RT-PCR和ELISA检测结果显示,与模型组肝组织中促炎细胞因子TNFα、IL-1β和IL-6 的mRNA(72.0±9.3、91.4±4.6、175.5±19.6)及蛋白表达量[(25.9±2.3)pg/mL、(816.5±60.8)pg/mL、(305.6±20.1)pg/mL]相比,干预组肝组织内促炎细胞因子TNFα、IL-1β和IL-6 的mRNA(15.4±0.2、6.0±1.6、21.5±0.9)及蛋白表达量[(7.9±1.2)pg/mL、(211.4±22.4)pg/mL、(53.2±7.3)pg/mL]均显著下调(P<0.05)。结论 DCH36_06对LPS/D-Gal诱导的小鼠急性肝损伤具有显著保护作用,其机制可能与下调肝内促炎细胞因子表达,减轻肝组织炎性损伤相关。

关键词: 急性肝损伤, DCH36_06, 细胞因子, 凋亡

Abstract: Objective To investigate the protective effect of histone acetyltransferase inhibitor DCH36_06 on mice with lipopolysaccharide (LPS)/D-galactose (D-Gal)-induced acute liver injury (ALI) and its mechanism. Methods A total of 75 C57BL/6 mice were randomly divided into 4 groups, 20 in normal control group, 20 in LPS/D-Gal-induced ALI group, 20 in DCH36_06 treatment group and 15 in DCH36_06 control group, injected intraperitoneally with normal saline, LPS/D-Gal, LPS/D-Gal + DCH36_06 and DCH36_06, respectively. After 4 hours of LPS/D-Gal injection, 5 mice in each of the first 3 groups were sacrificed. The peripheral serum and liver tissues of the mice were harvest to detect biochemical indexes, to evaluate liver tissue pathological changes and hepatocyte apoptosis by hematine-eosin and terminal deoxynucleotidyl transferase mediated biotinylated deoxyuridine triphosphate nick end labelling staining, and to detect the expression of pro-inflammatory cytokines in liver tissues by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The remaining mice were kept for 24 hours to test the 24-hour survival rate of each group. Results The 24-hour survival rate in the ALI group was only 40% (6/15), which reached 86.6% (13/15) in DCH36_06 treatment group (P<0.05). The necroinflammation and hepatocyte apoptosis of liver tissue were ameliorated in DCH36_06 treatment group compared with those in the ALI group. The serum levels of aspartate aminotransferase, alanine aminotransferase and total bilirubin in the treatment group were significantly lower than those in the ALI group (281.4±48.1 U/L vs 1151.0±111.1 U/L, 175.2±32.5 U/L vs 921.5±47.9 U/L, and 9.8±0.7 μmol/L vs 21.0±0.4 μmol/L, P<0.05). Additionally, the mRNA and protein levels in liver tissues of tumor necrosis factor alpha, interleukin 1β and interleukin 6 in the treatment group (72.0±9.3, 91.4±4.6, 175.5±19.6, 25.9±2.3 pg/mL, 816.5±60.8 pg/mL and 305.6±20.1 pg/mL) were significantly lower than those in the ALI group (15.4±0.2, 6.0±1.6, 21.5±0.9, 7.9±1.2 pg/mL, 211.4±22.4 pg/mL and 53.2±7.3 pg/mL) (P<0.05). Conclusion DCH36_06 can protect mice from LPS/D-Gal-induced ALI, and the mechanism might be related to the inhibition of crucial pro-inflammatory cytokines expression and liver inflammatory injury, indicating the epigenetic regulation could be a promising target for the treatment of ALI.

Key words: Acute liver injury, DCH36_06, Cytokine, Apoptosis

彭金金, 黄鹤鸣, 刘彦君, 石翠翠, 范建高, 张元元, 罗成, 李光明. 组蛋白乙酰化酶抑制剂(DCH36_06)对小鼠急性肝损伤的保护作用及机制研究[J]. 肝脏, 2019, 24(10): 1125-1128.

PENG Jin-jin, HUANG He-ming, LIU Yan-jun, SHI Cui-cui, FAN Jian-gao, ZHANG Yuan-yuan, LUO Cheng, LI Guang-ming. Protective effect and mechanism of histone acetyltransferase inhibitor DCH36_06 on mice with acute liver injury[J]. Chinese Hepatolgy, 2019, 24(10): 1125-1128.

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