Shen Mei-Jia, Sun Li-Chao, Liu Xiao-Yu, Xiong Meng-Chen, Li Shan, Tang A-Ling, Zhang Guo-Qiang
Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing 100193, China.
Emergency Department, China-Japan Friendship Hospital, Beijing 100029, China.
World J Emerg Med. 2022;13(3):182-188. doi: 10.5847/wjem.j.1920-8642.2022.056.
Sepsis-induced liver injury is a fatal complication of sepsis. Trichostatin A (TSA) regulates inflammation and autophagy in some human diseases, and forkhead box O3a (FoxO3a) has been shown to regulate autophagy. The present study aims to investigate whether TSA exerts its effects on septic liver injury through the FoxO3a/autophagy signaling pathway.
A sepsis mouse model was constructed by the cecal ligation and puncture (CLP) method, and AML12 cells were pretreated with lipopolysaccharide (LPS) (1 μg/mL) to establish a sepsis cell model. Forty mice were divided into four groups, namely control group, TSA group, CLP group, and CLP+TSA group, with 10 mice in each group. Cells were divided into control group, TSA group, LPS group, and LPS+TSA group. Hematoxylin-eosin (H&E) staining and biochemical methods were used to evaluate liver tissue injury. Enzyme-linked immunosorbent assay (ELISA) was applied to detect the expression of proinflammatory cytokines, and Western blotting and immunofluorescence were used to measure autophagy-related protein expression.
Compared with the CLP group (mice), the proinflammatory cytokines (interleukin-β [IL-β] 2,665.27±324.90 pg/mL to 2,080.26±373.66 pg/mL; interleukin-6 [IL-6] 399.01±60.98 pg/mL to 221.90±46.89 pg/mL) and the hepatocyte injury markers (aspartate transaminase [AST] from 198.18±27.07 U/L to 128.42±20.55 U/L; alanine aminotransferase [ALT] from 634.98±74.10 U/L to 478.60±32.56 U/L) were notably decreased after TSA intervention. Moreover, LC3 II and FoxO3a showed an obvious increase and P62 showed an obvious decrease in the CLP+TSA group. Cell experiment results showed the similar trend. After gene was knocked down in AML12 cells, the promotion of autophagy and the improvement of liver enzyme index and inflammation by TSA were weakened.
TSA may improve the inflammatory response and liver injury in septic mice through FoxO3a/autophagy.
脓毒症诱导的肝损伤是脓毒症的一种致命并发症。曲古抑菌素A（TSA）在一些人类疾病中调节炎症和自噬，并且已表明叉头框O3a（FoxO3a）可调节自噬。本研究旨在探讨TSA是否通过FoxO3a/自噬信号通路对脓毒症肝损伤发挥作用。
采用盲肠结扎穿孔（CLP）法构建脓毒症小鼠模型，并用脂多糖（LPS）（1μg/mL）预处理AML12细胞以建立脓毒症细胞模型。40只小鼠分为四组，即对照组、TSA组、CLP组和CLP + TSA组，每组10只小鼠。细胞分为对照组、TSA组、LPS组和LPS + TSA组。采用苏木精-伊红（H&E）染色和生化方法评估肝组织损伤。应用酶联免疫吸附测定（ELISA）检测促炎细胞因子的表达，并用蛋白质免疫印迹法和免疫荧光法检测自噬相关蛋白的表达。
与CLP组（小鼠）相比，TSA干预后促炎细胞因子（白细胞介素-β[IL-β]从(2,665.27±324.90) pg/mL降至(2,080.26±373.66) pg/mL；白细胞介素-6[IL-6]从(399.01±60.98) pg/mL降至(221.90±46.89) pg/mL）和肝细胞损伤标志物（天冬氨酸转氨酶[AST]从(198.18±27.07) U/L降至(128.42±20.55) U/L；丙氨酸转氨酶[ALT]从(634.98±74.10) U/L降至(478.60±32.56) U/L）显著降低。此外，CLP + TSA组中LC3 II和FoxO3a明显增加，P62明显减少。细胞实验结果显示出类似趋势。在AML12细胞中敲低基因后，TSA对自噬的促进作用以及对肝酶指标和炎症的改善作用减弱。
TSA可能通过FoxO3a/自噬改善脓毒症小鼠的炎症反应和肝损伤。
