ABSTRACT
Three compounds, including scolosprine C(1), uracil(2) and hypoxanthine(3), were isolated and purified from the ethyl acetate fraction of centipede by silica gel normal-phase column chromatography, reversed-phase medium pressure preparation chromatography, and high-pressure semi-preparative HPLC. The structure was elucidated through a combination of spectroscopic analyses [such as nuclear magnetic resonance(NMR) and mass spectrometry(MS)] and literature review. Among them, compound 1 was a new quinoline alkaloid. In previous reports, we have described the isolation and structure elucidation of one new and two known quinoline alkaloids. In this paper, we would report the isolation and structure elucidation of scolosprine C in detail.
Subject(s)
Animals , Alkaloids , Arthropods , Chilopoda , QuinolinesABSTRACT
Objective: Scolopendra was a traditional Chinese medicine(TCM) with a good medicinal value. Nowadays, there have been increasingly more adulterates of Scolopendra in the medicine market. To ensure the safe and effectiveness of clinical medicines,a convenient and accurate specific polymerase chain reaction(PCR) method for identification of medicinal Scolopendra from its common adulterates was established. Method: Based on the differences of COI gene DNA sequences among Scolopendra subspinipes mutilans and adulterants,the specific primer was designed,the reaction conditions were optimized,and the PCR method for identification was explored and verified in terms of tolerance and feasibility. Besides,the original animal samples and medicine of Scolopendra were collected. Result: Through the established PCR reaction system,the bright and simple fragments of 500 bp was amplified from DNA templates of S. subspinipes mutilans. All of the adulterants were negative by the multiplex PCR assay,such as S. multidens,S. subspinipes,S. dehaani,S. hainanum. Conclusion: The identified primer is highly specific,and the specific PCR method established in this paper can accurately identify Scolopendra and its adulterants, so as to provide an excellent scientific basis for the identification of TCM Scolopendra. The method is simple and intuitive, and facilitates wide promotion and application of the method, with a broad application prospect in the identification of TCM.
ABSTRACT
Objective: To study the quinoline alkaloids from the ethanol extract of Scolopendra subspinipes mutilans (SSM). Methods: The chemical constituents were isolated and purified by macroporous resin column, medium pressure preparation chromatography, and semi-preparative HPLC. Their structures were elucidated by IR, MS, and NMR experiments. Results: Three quinolone alkaloids were obtained and identified as 3-hydroxy-4-methoxyquinolin-8-yl hydrogen sulfate (1), jineol-8-sulfate (2), and jineol (3), respectively. Conclusion: Compound 1 is a new compound from SSM.
ABSTRACT
PURPOSE: The dried body of Scolopendra subspinipes mutilans has long been used as a traditional Korean medicinal food, but little is known about its mechanisms of action. In this study, we investigated the anti-inflammatory activities of Scolopendra subspinipes mutilans and possible mechanisms in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. METHODS: Cytotoxicity of Scolopendra subspinipes mutilans extract (SSME) was measured by MTT assay, anti-inflammatory activities were analyzed by nitric oxide (NO) production, the expression of inducible NO synthase (iNOS) and the mRNA level of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and interleukin-6 (IL-6). Nuclear translocation of nuclear factor-kappa B (NF-κB) p65 subunit and degradation of inhibitory kappa B (IκB) were examined by western blot. RESULTS: SSME inhibited LPS-induced NO production and iNOS expression without cytotoxicity. Up-regulation of LPS-induced pro-inflammatory cytokines, IL-1β and IL-6 was dose dependently attenuated by SSME. Exposure of pyrrolidine dithiocarbamate, an NF-κB specific inhibitor, accelerated the inhibitory effects of SSME on NO production and iNOS expression in LPS-stimulated cells. Moreover, translocation of NF-κB from the cytosol to the nucleus and degradation of IκB were decreased by treatment with SSME in LPS-induced cells. CONCLUSION: These results suggest that the SSME might have the inhibitory effects on inflammation, partly through inhibition of the NF-κB signaling pathway.
Subject(s)
Blotting, Western , Cytokines , Cytosol , Inflammation , Interleukin-6 , Nitric Oxide , Nitric Oxide Synthase , RNA, Messenger , Up-RegulationABSTRACT
Objective: To observe the effect of Scolopendra subspinipes extracts (SSE) on signal transducer and activator of transcription 3 (STAT3) signaling pathway and phosphorylation of its protein expression as well as the regulation mechanisms of HepG2 cells proliferation, invasion, and metastasis after SSE exposure. Methods: HepG2 cells were processed with SSE of gradient concentration (300, 600, 1 200, and 2 400 μg/mL). The inhibitory effect of SSE on HepG2 cell proliferation was evaluated by CCK8 method. Subsequent experimental concentration was set from IC50 result of CCK8 methods. HepG2 cells were divided into control, SSE (250 and 500 μg/mL), and 5-FU groups. After HepG2 cells were treated with SSE for 48 h, Transwell Chambers detected the invasion of HepG2 cells and Western blotting demonstrated expression and activation of STAT3, p-STAT3 and VEGF, MMP-2 protein. Results: CCK8 method showed that SSE had obvious inhibition effect on human HepG2 cells proliferation with dose dependent effect (P < 0.05). The IC50 of SSE was 508.3 μg/mL at 48 h. Transwell result showed invasive ability of human HepG2 cells was significantly reduced compared with control group after SSE worked to cells for 48 h (P < 0.05). Compared with 250 μg/mL SSE group, the number of membrane cells in 500 μg/mL SSE and 5-FU groups were less (P < 0.05). Western blotting analysis showed that STAT3 signaling pathway was mainly down regulated by p-STAT3 expression after SSE worked to cells for 48 h. Compared with control group, the p-STAT3 expression of SSE was down-regulated (P < 0.05). The MMP-2 and VEGF protein expression of 500 μg/mL SSE decreased compared with control group (P < 0.05). The MMP-2 protein expression of 500 μg/mL SSE group had obvious difference compared with the 250 μg/mL SSE group (P < 0.05). Conclusion: SSE regulate the activation of human HepG2 cells STAT3 signaling pathway by STAT3 phosphorylation down-regulation and reduce the expression of MMP-2 and VEGF downstream target protein, thus inhibited HepG2 cells proliferation, invasion, and metastasis ability in vitro.
ABSTRACT
Aim The aim is to purify fibrinolytic enzyme from Scolopendra subspinipes mutilans L.Koch and to study the thrombolytic and anticoagulant effect.Methods Scolopendra subspinipes mutilans L.Koch fibrinolytic enzyme(SSFE) was purified by ammonium sulphate precipitation,DEAE-cellulose and SephadexG-75 column chromatography from Scolopendra subspinipes mutilans L.Kochby.And fibrinolytic activity was determined by fibrin plate.The anticoagulant effect was measured on mice with haemolytic test and hemorrhagic test.The thrombolytic effect was measured with rats In vitro and in vivo,and the activated partial thromboplastin time(APTT),plasma prothrombin time(PT),thrombin time(TT) were measured.Results SSFE was single component with fibrinolytic activity and without any hemolyzation and hemorrhagic activity.All doses of SSFE(2,5,10 mg?kg-1) could obviously prolong activated partial thromboplastin time(APTT) and thrombin time(TT) ;Middle dose of SSFE(5 mg?kg-1) could prolong plasma prothrombin time(PT) while high dose of SSFE(10 mg?kg-1) didn't prolong obviously.Conclusion SSFE has obvious thrombolytic effect and anticoagulant effect.