1.Screening and Characterization of Effective Components of Immunopotentiating Activity in Senecionis cannabifolii Herba
Tingting ZHOU ; Difu ZHU ; Shengjun JI ; Chunchi WANG ; Dongxu JIA ; Yanru LI ; Yan TANG
China Pharmacy 2019;30(4):523-527
OBJECTIVE: To screen and characterize effective components of immunopotentiating activity in Senecionis cannabifolii Herba. METHODS: The polysaccharide components were obtained by water extraction and alcohol precipitation method to yield 50% alcohol precipitation sample (SCHE-1) and 80% alcohol precipitation sample (SCHE-2). The cells from mice mononuclear macrophage line RAW264.7 were divided into blank group (medium without serum), negative control group (medium with serum), lipopolysaccharide group (LPS, positive control drug, 1 μg/mL), SCHE-1 and SCHE-2 low-dose and high-dose groups (0.5, 1 mg/mL). The cell viability of RAW264.7 cells was detected by MTT assay. The levels of IL-1β, IL-6 and TNF-α in RAW264.7 were detected by ELISA. These were used to investigate the effects of SCHE-1 and SCHE-2 on the immunological enhancing activity of RAW264.7 cells. The molecular weight and distribution of SCHE-1 were determined by size exclusion chromatography; the monosaccharide composition of SCHE-1 was determined by HPLC pre-column derivatization. Methylation analysis of SCHE-1 was conducted by NaOH method. RESULTS: Compared with negative control group, the activity of RAW264.7 cells was enhanced significantly in SCHE-1 groups and LPS group, which also significantly increased the levels of IL-1β, IL-6 and TNF-α in cell culture fluids (P<0.01). SCHE-1 was an effective component with immunopotentiating activity. The neutral sugar content of SCHE-1 was 40.05%, the uronic acid was 35.62%, and the protein was 8.89%. SCHE-1 was a mixture, molecular weight of which was 62-6 119 Da; monosaccharide was mainly composed of galacturonic acid, arabinose (Ara) and galactose (Gal). The results of methylation analysis showed that the backbone was composed of 1→3, 1→4 and 1→6 linked Gal, and branches were on the O-6 position of the 1→3 linked Gal, and the non-reducing terminals were Ara. CONCLUSIONS: SCHE-1 may be the effective component of immuno potentiating activity, and main component of SCHE-1 is polysaccharide. SCHE-1 may regulate the immune function by activating macrophages to release IL-1β, IL-6 and TNF-α.