Indole-3-Carbinol Derivative DIM Mitigates Carbon Tetrachloride-Induced Acute Liver Injury in Mice by Inhibiting Inflammatory Response, Apoptosis and Regulating Oxidative Stress

Abstract

3,3′-Diindolylmethane (DIM), a metabolic product of indole-3-carbinol extracted from cruciferous vegetables exhibits anti-inflammatory and anti-cancer properties. Earlier, the product has been demonstrated to possess anti-fibrotic properties; however, its protective effects on liver injury have not been clearly elucidated. In this study, we postulated the effects and molecular mechanisms of action of DIM on carbon tetrachloride (CCl4)-induced liver injury in mice. Acute liver injury was induced by a single intraperitoneal administration of CCl4 (1 ml/kg) into mice. DIM was injected via subcutaneous route for three days at various doses (2.5, 5 and 10 mg/kg) before CCl4 injection. Mice were sacrificed and serum was collected for quantification of serum transaminases. The liver was collected and weighed. Treatment with DIM significantly reduced serum transaminases levels (AST and ALT), tumor necrosis factor-α (TNF-α) and reactive oxygen species (ROS). CCl4- induced apoptosis was inhibited by DIM treatment by the reduction in the levels of cleaved caspase-3 and Bcl2 associated X protein (Bax). DIM treated mice significantly restored Cytochrome P450 2E1, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression in CCl4 treated mice. In addition, DIM downregulated overexpression of hepatic nuclear factor kappa B (NF-κB) and inhibited CCl4 mediated apoptosis. Our results suggest that the protective effects of DIM against CCl4- induced liver injury are due to the inhibition of ROS, reduction of pro-inflammatory mediators and apoptosis.

Polydeoxyribonucleotide Exerts Protective Effect Against CCl4-Induced Acute Liver Injury Through Inactivation of NF-κB/MAPK Signaling Pathway in Mice

“…Serum activity such as ALT and AST is a very sensitive marker used in the diagnosis of liver disease [ 1 ]. In particular, an increased level of serum AST and ALT is a major indicator of CCl 4 -induced liver toxicity with cell membrane damage and loss of hepatocyte functional integrity [ 10 ].…”

“…Induction of excessive reactive oxygen species by CCl 4 activates Kupffer cells, which mediate liver inflammatory processes through secretion of TNF-α, IL-1β and IL-6 [ 16 , 26 ]. Overproduction of pro-inflammatory cytokines causes inflammation-related liver damage and plays a critical role in the development process [ 10 ]. In the current results, increased secretion of TNF-α, IL-1β and IL-6 in serum and liver tissue is due to the damaging effect of CCl 4 .…”

“…Excessive apoptosis exacerbates liver damage, and treatment of acute liver disease requires relief of apoptosis [ 10 ]. The ratio of Bax vs. Bcl-2 is an important determinant for apoptosis, and DNA fragmentation after exposure to CCl 4 was observed in liver tissue [ 22 ].…”

“…These inflammatory cells secrete pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6, and these pro-inflammatory cytokines play a role in promoting ALI progression. During ALI progression, various damaging factors contribute to the inflammatory response and promote liver cell apoptosis, thereby increasing liver tissue damage [ 9 , 10 ].…”

Anticancer effects of Poncirus fructus on hepatocellular carcinoma through regulation of apoptosis, migration, and invasion

“…The cells were washed with PBS and lysed with lysis buffer (iNtRON Biotechnology) containing a cocktail of phosphatase inhibitor-1. Western blotting was performed as previously described ( 38 ). The membranes were incubated overnight with primary antibodies specific for anti-rabbit cleaved caspase-3 (cat.…”