Development of phytosterol loaded silver nanoparticles for ameliorating hemorrhoidal complications via AMPK pathway- a mechanistic approach.

The current study was aimed to synthesize silver nanoparticles (PLSNPs) using green technology by means of phytosterol enriched fractions (EAF) from Blumea lacera extracts and evaluate its toxicological and anti-hemorrhoidal potential. The peak at 410 nm wavelength in the UV-Visible spectrophotometer verified the fabrication of PLSNPs because of surface plasmon resonance. Average size of synthesized particle was found to be 85.64 nm supported by SEM and TEM. EDS analysis showed the elemental composition of synthesized PLSNPs having carbon (C) 12.59% besides silver (Ag) 87.41%, indicating the capping of phytochemicals on the PLSNPs. The PLSNPs were also standardized for total phytosterol content using chemical and HPLC methods. The PLSNPs were found to be safe upto 1000 mg/kg as no toxicity was observed in the acute and sub-acute toxicity study performed as per OECD guidelines. After inducing haemorrhoids, the experimental animals were treated with different doses of EAF, PLSNPs and standard drug (Pilex) for seven days and on eighth day, the ameliorative potential was assessed by evaluating the hemorrhoidal, biochemical parameters (TNF-α and IL-6) and histology of recto-anal tissue. The results showed that treatment with PLSNPs and Pilex significantly (P<0.05) reduced haemorrhoidal (inflammatory severity index, recto-anal coefficient) and biochemical parameters. This was further supported by restoration of altered antioxidant status. Further, a marked reduction in the inflammatory zones along with minimal dilated blood vessels were observed in histopathological study. The results of molecular docking studies also confirmed the amelioration of haemorrhoids via AMPK mediated reduction of inflammation and endothelin B receptor modification by PLSNPs.