Efficient, accurate and comprehensive evaluation of polysaccharides from Fritillaria and their inhibitory responses to mouse inflammation

Abstract

Fritillaria is a perennial herb with bulbs with multiple medicinal usages. Here, we optimized extraction and hydrolysis methods for different species and origins of Fritillaria to study their anti-inflammatory activities. Using ultrasound-assisted hot water extraction after hydrolysis by hydrochloric acid and trifluoroacetate (TFA), monosaccharide derived from 1-phenyl-3-methyl-5-pyrazolone (PMP) was separated by using HPLC. The optimal conditions for hot water extraction were found via single factor analysis and orthogonal experiments. The PMP derivatization HPLC method could accurately determine the Mannose (Man), Glucose (Glu), Galactose (Gal), Xylose (Xyl), and Fucose (Fuc) content values in Fritillaria. This pre-column derivatization method is simple and rapid, providing less variation, high sensitivity and good reproducibility for saccharide separation. To study Fritillaria's anti-inflammatory effects in a mouse swelling model, HE-staining was performed to observe morphological changes in liver and kidney samples. Fritillaria's active ingredients could significantly inhibit mouse ear swelling induced by xylene and inhibit toe swelling induced by egg white. They mainly inhibited the secretion of inflammatory cytokines inside the body and maintained the dynamic balance between pro-inflammatory and anti-inflammatory factors in the body. According to histopathological analysis, Fritillaria had good anti-inflammatory activity against LPS-induced inflammation, reducing the expression of inflammatory cytokines.