Hepatoprotective and anti-carcinogenic potential of mercury-containing drug, Lokanatha Rasa against hepatocellular carcinoma: In vitro cytotoxic, anti-inflammatory and genotoxic evaluation

Hepatocellular carcinoma (HCC) is one of the most prevalent and lethal malignancies worldwide and it is one of the top causes of cancer-related deaths. It is also closely linked to chronic inflammation, oxidative damage and persistent hepatocellular injury, which are all factors that contribute to the development of cancer. The conventional treatments for cancer, such as chemotherapy, radiation therapy and targeted biologic therapies, are often plagued by systemic toxicity, resistance and poor response rates, thus underlining the need for multi-targeted therapies. Rasa Shastra is a specialized branch of Ayurveda that lists classical herbo-mineral and mercurial compounds for the treatment of liver ailments (Yakrit Vikara) and chronic metabolic disorders. Lokanatha Rasa (LKN) is a classical mercurial drug that is made up of purified mercury (Shoditha Parada), sulphur (Shoditha Gandhaka), calcined minerals, and cowry shells, which are further processed by classical methods of purification (Shodhana) and incineration (Marana). This work assessed the in vitro cytotoxic, anti-inflammatory and genotoxic properties of LKN on HepG2 human hepatocellular carcinoma cells. Cytotoxicity was assessed by the MTT assay, anti-inflammatory activity by the Human Red Blood Cell (HRBC) membrane stabilization assay and genotoxicity by the comet assay. LKN exhibited dose-dependent cytotoxicity with an IC₅₀ of 1.6 mg/mL, while anti-inflammatory activity increased with concentration, although diclofenac had higher membrane-stabilizing activity. The comet assay showed DNA strand breaks at medium and high concentrations, indicating possible apoptosis induction. The combined results suggest that LKN has multi-targeted biological activity, which includes moderate cytotoxicity, membrane-stabilizing anti-inflammatory activity, and DNA damage. These findings offer preliminary scientific evidence for the traditional use of Rasa Shastra drugs in liver diseases and emphasize the importance of traditional processing methods in improving their efficacy. LKN has potential as an adjuvant in HCC treatment and requires further investigations.