Isolation of Saponins with the Inhibitory Effect on Nitric Oxide, Prostaglandin E2 and Tumor Necrosis Factor-.ALPHA. Production from Pleurospermum kamtschaticum

Abstract

Pleurospermum kamtschaticum HOFFMANN (Umbelliferae) is a perennial herb distributed in Kangwon province Korea.1)The aerial part of P. kamtschaticum are traditionally used to treat colds, arthritis, atherosclerosis and impotence.2) The constituents of P. kamtschaticum have not been reported though Luo et al. 3) reported on the isolation of saponins from another Pleurospermum spp. Activity-directed fractionation led to the isolation of the two saponins compounds 1 and 2 from P. kamtschaticum, which were identified as saikogenin (buddlejasaponin IV) and 3b,16b,23, by physicochemical and spectroscopic data. The structures of the two saponins are shown in Fig. 1. Compounds 1 and 2 have not been previously isolated from Pleurospermum spp. These two compounds were tested for their inhibitory effects on nitric oxide (NO) production in an lipopolysaccharide (LPS)-stimulated macrophage cell line (Raw 264.7 cells). NO has diverse physiological roles and also contributes to immune defenses against viruses, bacteria, and other parasites. However, excessive NO production is associated with various diseases such as arthritis, diabetes, stroke, septic shock, autoimmune diseases, chronic inflammation, and atherosclerosis. 4)Previously, we reported on the cytotoxic effects of triterpenes and triterpenoidal saponins against cancer cell lines and on the inhibition of NO production in LPS-activated macrophage cell lines. 5-7)In general, sapogenins are bioactive principles rather than the saponins themselves. Therefore, saikosapogenin A (3) and -H (4), which are artifacts rather than genuine aglycones, were obtained from the hydrolysis of the two isolated saponins. However, it was found that these significant NO inhibitory effect of two sapogenins was due to their cytotoxic effect on RAW 264.7 cells. Thus, as a prelude to reveal the underlying mechanisms for the anti-inflammatory effect of compounds 1 and 2, we evaluated and compared the effects of these compounds isolated from the aerial portion of P. kamtschaticum on LPS-induced NO, prostaglandin E 2 (PGE 2 ) and tumor necrosis factor-a (TNF-a) release by the macrophage cell line RAW 264.7 in the present study. MATERIALS AND METHODSInstruments Melting Points were determined on an Electrothermal digital melting point apparatus. Optical rotations were measured on a JASCO DIP-360 digital polarimeter and IR spectra recorded on a Bomem MB-100 FT-IR spectrometer. FAB-MS were obtained using a JMS HX-110 As an attempt to search for bioactive natural products exerting antiinflammatory activity, we have isolated two saponins were isolated from the aerial portion of Pleurospermum kamtschaticum (Umbelliferae) by nitrite assay activity-directed chromatographic fractionation. They were identified as saikogenin . Compound 1 significantly inhibited nitric oxide (NO) production, and it also significantly decreased prostaglandin E 2 (PGE 2 ) and tumor necrosis factor-a a (TNF-a a) release in the lipopolysaccharide (LPS)-activated macrophage Raw 264.7 cells whereas compound 2 was much less active....

Pleurospermum kamtschaticumExtract Induces ApoptosisviaMitochondrial Pathway and NAG-1 Expression in Colon Cancer Cells

“…PKE, including 0.93 g of buddlejasaponin IV/100 g PKE, was generously provided by Professor Hee-Jun Park, a coauthor of this work. 3) The following antibodies were purchased from the following sources: Bax, Bcl-2, procaspase-9, and horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA); PARP and procaspase-3 (Cell Signaling Technology, Beverly, MA); NAG-1 (Upstate Biotechnology, Lake Placid, NY); and -actin (Sigma, St. Louis, MO).…”

“…2) Additionally, isolated buddlejasaponin IV evidenced potent anti-inflammatory activity, blocking the production of nitric oxide, prostaglandin E 2 , and tumor necrosis factor-in lipopolysaccharide-stimulated Raw 264.7 cells. 3) Colorectal cancer remains one of the most frequent malignant neoplasms worldwide. Epidemiological studies have suggested that the incidence and progression of colorectal cancer can be prevented by dietary factors, most notably a high intake of fruits and vegetables.…”

Anti‐inflammatory effect of buddlejasaponin IV through the inhibition of iNOS and COX‐2 expression in RAW 264.7 macrophages via the NF‐κB inactivation

“…During our on‐going screening program designed to identify the anti‐inflammatory potentials of natural compounds, we previously isolated buddlejasaponin IV from the aerial portion of Pleurospermum kamtschaticum by activity‐directed fractionation and characterized its structural identity spectroscopically ( 1 H and 13 NMR, IR, MS), as described previously (Jung et al ., 2005). P. kamtschaticum Hoffmann (Umbelliferae) is a perennial herb and is found in Kangwon province Korea (Koh & Jeon, 2003).…”

Preoperative Oral Administration of Kikyo-To, a Kampo Medicine, Alleviates Postoperative Sore Throat: A Prospective, Double-Blind, Randomized Study

“…13 In addition, saponin, a component of KKT, has been reported to reduce the level of TNF-a or prostaglandin E2, which is a key molecule involved in inflammatory pain. 14,15 These reports suggest that KKT might be beneficial in treating inflammation-related pain.…”