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Screening for Protective Effect Target of Deproteinized Extract of Calf Blood and Its Mechanisms in Mice with CCl4-Induced Acute Liver Injury

ABSTRACT: Liver injury is the common pathological basis of various liver diseases, and its existence for a long time is often a important initiation factor to lead to liver fibrosis, and even liver cirrhosis and hepatocellular carcinoma (HCC), so that the prevention and treatment of liver injury is one of the key links in the clinical therapy of liver diseases, and the timely control of the occurrence and development of liver injury will have an important clinical significance in the treatment of liver disorders. It has been reported that deproteinized extract of calf blood (DECB) can inhibit the replication of hepatitis B virus and has a protective effect on the liver function after a traumatic liver injury. However, there are few studies on the regulatory factors and mechanisms of DECB effects. In this study, an acute mouse liver injury model was established with carbon tetrachloride (CCl4), mRNA expression microarray was used to screen the differentially expressed genes and related pathways in mice in the DECB group and model group, and the target of protective effect of DECB and its related mechanism were found in mice with acute liver injury induced by carbon tetrachloride, which may provide an important theoretical basis for the further research and development of DECB.

ORGANISM(S): Mus musculus

PROVIDER: GSE82771 | GEO | 2017/09/18

SECONDARY ACCESSION(S): PRJNA324828

REPOSITORIES: GEO

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Project description:Since xenobiotics enter the organism via the liver, hepatocytes must cope with numerous perturbations. In particular, exposure to hepatotoxic agents resulting in liver injury triggers a strong alteration in gene expression. The contribution of these transcriptional regulatory networks to the propagation of injury (i.e. by modulation of metabolic pathways, inflammation, proliferation) are not fully understood. Therefore, we conducted a time-resolved gene array study on mouse liver after exposure to a single intraperitoneal injection of the hepatotoxic compound carbon tetrachloride (CCl4). This model induces a transient injury which reaches its maximum between 2 to 3 days after injection, followed by a precise regenerative response. The experiment included early time points (i.e. 2 and 8h) and late time points (1 to 16 days) to cover the initial events induced by CCl4 metabolization and cell stress, and the regenerative phase between 2-6 days after intoxicaiton. C57BL/6N male mice (8-12 weeks old) were obtained from Charles Rivers (Sulzfeld, Germany). The local Ethics committee approved the experiments. 4-5 mice were used for each time point. Mice received a single intraperitoneal injection of CCl4 (1.6 g/kg body weight) dissolved in 0.5 ml olive oil. The liver tissue was collected after 2h, 8h, and 1, 2, 4, 6, 8 and 16 days following CCl4 administration. As control, mice received 0.5 ml of olive oil intraperitonealy for 1 or 8 days. Healthy liver was collected from untreated mice. At the indicated time points, mice were anesthetized and blood was collected from the heart with a 25 gauge needle, using EDTA as anticoagulant, for analysis of the liver transaminases GOT and GPT in serum. Afterwards, the liver was resected, washed in ice cold PBS and sectioned for further analysis. Liver tissue sections of about 0.5 cm2 were snap frozen in liquid nitrogen and stored at -80ﾰC for subsequent isolation of proteins or RNA. RNA was isolated from mouse liver tissue using the Phenol/Chloroform method (Trizolﾮ, Qiagen, Hilden, Germany) according to the manufacturerﾒs instructions. RNA concentration and integrity were determined spectrophotometrically in a Nanodropﾮ2000 (ThermoScientific, Waltham MA, USA) and in a Bioanalyzerﾮ (Agilent, Waldbronn, Germany), respectively.

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Screening for Protective Effect Target of Deproteinized Extract of Calf Blood and Its Mechanisms in Mice with CCl4-Induced Acute Liver Injury

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