Supplementary material from "Band Gap Energy shift as a Biophysical Marker of Puerarin’s Neuroprotective Role in Epilepsy-Induced Demyelination "

The study aims to determine the neuroprotective effect of Puerarin, extracted from Pueraria Lobata roots, against Pentylenetetrazole (PTZ)-induced epileptic seizures and concomitant demyelination and low conductance through Diffuse Reflectance Spectroscopic (DRS) technique. Mice groups (n=12) were pretreated with vehicle and Puerarin (100 mg/Kg, intraperitoneal (i.p.)) for 42 days (alternate days). PTZ (35 mg/Kg, i.p.) was given after 30 minutes to all groups except to the vehicle control group. Puerarin remarkably reduced seizures scoring, hyperaglesia and blood brain barrier disruption, and improved muscle coordination. High anti-Myelin Oligodendrocyte Glycoprotein (anti-MOG) levels in PTZ-treated mice confirmed demyelination, with low molar absorption coefficient (µa) and high scattering coefficient (µs) and absorption. Puerarin treatment increased µa and decreased anti-MOG and µs, similar to vehicle control group. This indicates protective effect of Puerarin against demyelination. Band gap energy (Ebg) calculated by Kubelka-Munk model and incorporated in current study first time to evaluate pathology. Ebg Blue shift showed compromised conductance capacity of nerves in PTZ-induced epilepsy than Puerarin treated animals. Hematoxylin and Eosin (H & E) and nissl staining were used to further authenticate the neuroprotective effect of Puerarin. In conclusion, Puerarin countered demyelination and low conductance of nerve due to PTZ induced epilepsy assessed via DRS.