Study on the anti-inflammatory effect of 4’-O-β-D-glucosyl-5-O-methylvisamminol in Radix Saposhnikoviae on lipopolysaccharide -induced RAW 264.7 cell injury#br#
MENG Li1 ZHENG Wei2 LIU Pengpeng2 JIA Tianzhu2
1.Liaoning Traditional Chinese Medicine Institute, Liaoning Province, Shenyang 110034, China;
2.College of Pharmacy, Liaoning University of Traditional Chinese medicine, Liaoning Province, Shenyang 110032, China
Abstract：Objective To study the protective effect and mechanism of 4’-O-β-D-glucosyl-5-O-methylvisamminol in Radix Saposhnikoviae on inflammation of mice RAW264.7 cells induced by lipopolysaccharide (LPS). Methods RAW264.7 mononuclear macrophages were induced cellular inflammatory model by LPS. The proliferation effect of 4’-O-β-D-glucosyl-5-O-methylvisamminol with different concentrations(160, 80, 40, 20, 10 μmol/L) on mice RAW264.7 cells was detected by MTT assay. The experiment was divided into blank group, LPS group, positive control group (Indomethacin, 10 μmol/L), low (20 μmol/L), medium (40 μmol/L) and high (80 μmol/L) dose groups of 4’-O-β-D-glucosyl- 5-O-methylvisamminol. Blank medium was added to blank group and LPS group, and corresponding drugs were added to other dosing groups. LPS (10 μg/ml) was added after 2 h, the secretion of nitric oxide (NO) in supernatant of mice RAW264.7 was detected by Griess reagent method, the contents of prostaglandin E2 (PGE2) and tumor necrosis factor α (TNF-α) in the supernatant of mice RAW264.7 cells were determined by enzyme linked immunosorbent assay. The mRNA expressions of nuclear factor κB (NF-κB p65), interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were detected by RT-PCR. Toll like receptor 4 (TLR4) and NF-κB p65 protein expressions were detected by Western blot. Results 4’-O-β-D- glucosyl-5-O- methylvisamminol (10-80 μmol/L) had no effect on RAW264.7 cell viability. Compared with LPS group, the contents of NO, PGE2 and TNF-α in mice RAW264.7 cells were decreased in low, medium and high doses of 5-O-methylvisamminol side groups (P＜0.01). The mRNA expressions of NF-kB p65, IL-1β, INOS and COX-2 and the protein expressions of TLR4 and NF-kB p65 were decreased (P＜0.01). Conclusion 4’-O-β-D-glucosyl-5-O- methylvisamminol in Radix Saposhnikoviae inhibit the inflammation of RAW264. 7 cells induced by LPS has anti- inflammatory effect.
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