Chebulinic Acid Ameliorates Diethylnitrosamine-Induced Hepatocarcinogenesis in Rats by Inhibiting AKT/mTOR Activation and Inhibits Liver Cancer Cell (HepG2) Proliferation

Hepatocellular carcinoma is an aggressive cancer with a global impact, exacerbated by the increasing rates of metabolic-dysfunction-associated steatotic liver disease and alcohol-related liver disease. Although recent years have seen a stabilization in hepatocellular carcinoma incidence due to effective viral hepatitis treatments and advances in early detection and therapy, the overall burden is projected to rise. Hepatocellular carcinoma treatment requires a multidisciplinary approach since it depends on the tumor stage, patient performance status, and liver function. Significant progress in surgical and locoregional therapies has improved short-term survival rates; however, disease recurrence remains a major challenge. Hence a drug that effectively attenuates carcinogenesis in hepatocytes and prevents the recurrence of cancer cells is required to treat hepatocellular carcinoma. Chebulinic acid, an ellagitannins phytochemical, possess varied pharmacological properties and acts as a multi-targeted antitumor drug with significant therapeutic potential. We evaluated the potency of chebulinic acid attenuating diethylnitrosamine-induced hepatocarcinoma in an animal model. Hepatocellular carcinoma was induced in rats by feeding with potable water consisting of 0.01% diethylnitrosamine for 12 weeks. Chebulinic acid was given to hepatocellular carcinoma-induced rats for 8 weeks. The feed intake and weight gain of rats were assessed periodically during the treatment period. Hepatic enzymes and the antioxidants levels were quantified in the experimental animals to evaluate the hepatoprotective effect of chebulinic acid. Alpha-fetoprotein and carcinoembryonic antigen protein levels were measured to confirm the induction of carcinogenesis and the anticancer potency of chebulinic acid. Cytokines stimulating inflammation were measured to analyze the anti-inflammatory potency of chebulinic acid against diethylnitrosamine-induced inflammation in hepatocytes. The apoptotic and tumor suppressing properties of chebulinic acid were evaluated via quantifying the apoptotic stimulating proteins and tumor suppressor proteins in diethylnitrosamine-treated rats. AKT/mTOR proteins were quantified to assess the attenuating effect of chebulinic acid against diethylnitrosamine-induced abnormal cell proliferation. The anticancer potency of chebulinic acid was confirmed with hepatic histopathological analysis. In in vitro assay, the effect of chebulinic acid was studied against HepG2 cells. Chebulinic acid treatment effectively regulated the hepatic enzymes, increased the antioxidant status, triggered apoptosis, and stimulated tumor suppressor genes in diethylnitrosamine-treated rats. It also effectively attenuated inflammation and cell proliferation signaling in hepatocellular carcinoma-induced rats. The overall results of our investigations prove chebulinic acid is a multi-targeted anticancer drug that effectively ameliorated diethylnitrosamine-induced hepatocellular carcinoma in rats.