Vol. 1, Issue 1, Part A (2024)

Background: Tinospora cordifolia (Willd.) Miers, a traditional medicinal plant extensively used in Ayurvedic medicine, has demonstrated various pharmacological activities, including anticancer potential. Among its phytochemical constituents, alkaloids have been associated with potent biological effects. This study aimed to evaluate the cytotoxic and apoptotic effects of the alkaloid-rich fraction of T. cordifolia stems on HeLa cervical cancer cells.

Methods: Alkaloids were extracted using acid-base fractionation from dried stem powder and characterized via phytochemical screening and FTIR analysis. HeLa cells were cultured and treated with various concentrations (25-400 µg/mL) of the extract for 24 and 48 hours. Cell viability was assessed using MTT assay, and apoptotic morphology was observed using phase-contrast microscopy. Nuclear changes were analyzed via DAPI staining. Apoptotic induction was confirmed using Annexin V-FITC/PI flow cytometry, and caspase-3 activity was measured colorimetrically. Data were analyzed using one-way ANOVA and t-tests with significance at p<0.05.

Results: The alkaloid fraction reduced HeLa cell viability in a dose- and time-dependent manner, with IC₅₀ values of 132.5 µg/mL (24h) and 91.3 µg/mL (48h). Microscopic analysis revealed features consistent with apoptosis. DAPI-stained cells showed chromatin condensation and nuclear fragmentation. Flow cytometry revealed a substantial increase in early and late apoptotic populations, while caspase-3 activity increased nearly threefold compared to control. Statistical analysis confirmed the significance of these findings (p<0.001).

Conclusion: The alkaloid fraction of T. cordifolia exhibits strong cytotoxic effects on HeLa cells by inducing apoptosis, primarily through caspase-3-mediated pathways. These results highlight the therapeutic potential of this natural extract and recommend further in vivo and mechanistic studies for clinical translation. The study underscores the value of herbal alkaloids in developing cost-effective anticancer treatments.