Phyllanthus emblica: biointeraction between seed methanolic extract and glioma cells

This study attempts the scientific validation of the neuroprotective biointeraction of Phyllantus emblica (PE) seed methanolic extract (PESME) against C6 glioma cells. PE fruit pulp and seeds were sequentially extracted, and the free radical scavenging activity of extracts was screened on primary mouse splenocytes and C6 glioma cells using DPPH and MTT assays. PESME was chemically profiled using HPTLC-densitometry. PESME treated C6 cells were subjected to morphological and lysosomal membrane integrity analyses using Giemsa, Coomassie brilliant blue and acridine orange staining. Oxidative stress parameters like lipid peroxidation, glutathione and antioxidant enzymes were assessed. Reactive oxygen species (ROS) and cell proliferative potential were evaluated by DCFDA, click EdU and cell cycle analyses. PESME was evaluated in vivo for short term acute toxicity and histopathology. PESME (50 µg/mL) significantly alleviated the prooxidant properties of C6 cells by reducing the levels of oxidative stress parameters. Besides it inhibited the generation of ROS in C6 cells, with only 25.2% of DCFDA^+ve cells detected after the treatment. Moreover, PESME inhibited C6 cell growth at the 'S' and 'G2' phases, indicating a barrier to DNA synthesis. This suggests that PESME may decrease mitosis of C6 cells. Gross and histopathological evaluation of brain and liver revealed normal architecture in both control and PESME treated groups suggesting no organ and tissue toxicity. The results of this study demonstrate the capacity of PESME to inhibit C6 cell proliferation and its potential application as a therapeutic agent.