Research on Protective Effect and Mechanism of Idazoxan on lps Attacked Acute Hepatic Injury

2017 3rd International Conference on Environmental Science and Material Application

生态环境科学;材料科学

Zhu, Junyu^1 ; Ying, Shangqi^2 ; Kang, Wenyuan^3 ; Huang, Wenjuan^4 ; Liang, Huaping^1

State Key Laboratory of Trauma, Burns and Combined Injury, Research Institute of Surgery, Daping Hospital, Third Military Medical University, Chongqing

400042, China^1

Department of Clinical Laboratory, Affiliated Hospital of Chongqing, Medical University, Chongqing

400016, China^2

State Key la (Boratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha

410082, China^3

Department of Severe Medicine, Hospital of Zunyu, Medical College, Zunyi

563000, China^4

关键词: Aspartate aminotransferase;    Concentration dependence;    Enzyme linked immunosorbent assay;    Inflammatory mediators;    Intraperitoneal injections;    MRNA expression level;    Reactive oxygen species;    Tumor necrosis factor alpha;   

Others  :  https://iopscience.iop.org/article/10.1088/1755-1315/108/4/042089/pdf
DOI  :  10.1088/1755-1315/108/4/042089

来源: IOP

【 摘 要 】

Objective: To observe the protection effect of Idazoxan (IDA) on LPS induced acute hepatic injury, and to explore its action mechanism. Methods: 60 adult C57BL/6 mice were divided into a control group (20 mice, intraperitoneal injection of phosphate buffer), a model group (20 mice, intraperitoneal injection of LPS 10 mg/kg) and a agmatine group (20 mice, intraperitoneal injection of LPS 10 mg/kg and agmatine 200 mg/kg) according to random number table method. Blood and liver tissue were collected for preparation of tissue homogenate. Enzyme-linked immunosorbent assay (ELISA) was adopted for detecting tumor necrosis factor-α (TNF-α) and interleukin (IL- 1β and IL - 6) contents in the serum and liver tissue at 24h after molding. Automatic biochemical analyzer is used for determining alanine transaminase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) level at 24h after molding; The liver tissue pathology changes were observed at 24h after molding. Macrophage RAW264.7 cells were stimulated by 10 μg/mL LPS and with or without IDA (100 μmol/L). 2', 7'-dichlorofluoresce in diacetate (DCFH-DA) was used as a fluorescent probe for detection of intracellular reactive oxygen species (ROS) level; qRT - PCR method was used for detecting antioxidant enzymes HO-1 and NQO-1 mRNA expression level at 2h, 4h and 8 h. Results: mice in the model group suffered from depression, curling and food water forbidding at 6h after molding. Mice in the Idazoxan group have obviously better spirit and activity than that of model group. The serum ALT, AST and LDH level of LPS attacked acute hepatic injury mice can be effectively alleviated after Idazoxan treatment. The expression of proinflammatory factor TNF-α and IL-6 in the liver can be reduced. The liver showed obvious pathological changes at 24 h after injection, such as liver cell swelling, necrosis, congestion, inflammatory cell infiltration, etc.; The liver cell injury was prominently alleviated in IDA treatment group. Compared with the control group, LPS significantly increased ROS level in RAW264.7 cells. The ROS level was decreased with concentration dependence after IDA intervention. IDA increased HO-1 mRNA expression of RAW264.7 cells. It had no influence on NQO-1mRNA. Conclusion: IDA significantly reduces the serum liver injury indexes and contents of TNF-α, IL-6 and other inflammatory mediators in liver tissues. It can alleviate the liver pathology change, thereby it can generate protection function on LPS attacked acute hepatic injury. Its action mechanism may be related to IDA-enhanced liver macrophage antioxidant function.