RESUMO
The gene fragment of PFC0460w was amplified from RNA of Plasmodium falciparum 3D7 strain with RT-PCR, and cloned into pGEM-T easy vector. The recombinant plasmid was transformed into E. coli DH5alpha and the positive clones were selected, which were identified by PCR and sequencing. The results showed that there were three sequences of PFC0460w fragment, respectively with length of 618, 597 and 543 bp. The 618 bp fragment was completely consistent with the sequence published in PlasmoDB (GenBank No. XM_001351147), and the 597 bp and 543 bp fragments were submitted to GenBank with Accession No. of JF799872 and JF799873, respectively. 205 amino acids were encoded by the 618 bp fragment, and five kinds of protein structure were predicted by Robetta.
Assuntos
Biologia Computacional , Genes de Protozoários , Plasmodium falciparum/genética , Animais , Clonagem Molecular , Vetores Genéticos , Dados de Sequência Molecular , PlasmídeosRESUMO
Many species from Senecio genus have been used in traditional medicine, and their pharmacological activities have been demonstrated. This study investigated the chemical composition and anti-inflammatory activities of essential oils from Senecio flammeus. A total of 48 components representing 98.41 % of the total oils were identified. The main compounds in the oils were α-farnesene (11.26 %), caryophyllene (8.69 %), n-hexadecanoic acid (7.23 %), and α-pinene (6.36 %). The anti-inflammatory activity of the essential oils was evaluated in rodents (10-90 mg/kg bw) in classical models of inflammation [carrageenan-induced paw edema, 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced ear edema, and cotton pellet-induced granuloma]. The essential oils at doses of 10, 30, and 90 mg/kg bw significantly reduced carrageenan-induced paw edema by 17.42 % (P < 0.05), 52.90 % (P < 0.05), and 66.45 % (P < 0.05) 4 h after carrageenan injection, respectively, and significantly reduced myeloperoxidase activity (P < 0.05). The essential oils (10, 30, and 90 mg/kg) also produced a significant dose-dependent response to reduce TPA-induced ear edema by 20.27 % (P < 0.05), 33.06 % (P < 0.05), and 53.90 % (P < 0.05), respectively. The essential oils produced significant dose-response anti-inflammatory activity against cotton pellet-induced granuloma that peaked at the highest dose of 90 mg/kg (49.08 % wet weight and 47.29 % dry weight). Results demonstrate that the essential oils of S. flammeus were effective in the treatment of both acute and chronic inflammatory conditions, thereby supporting the traditional use of this herb.
RESUMO
Selagin-7-O-(6''-O-acetyl-)-ß-D-glycoside, a new flavone glycoside isolated from Cancrinia discoidea, is known to exhibit anti-inflammatory activity in vivo. This study aimed to investigate the protection of this flavone glycoside on inflammation in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The effects of selagin-7-O-(6''-O-acetyl-)-ß-D-glycoside on inflammatory cytokines and signaling pathways were analyzed by enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, and western blot. Results show that selagin-7-O-(6''-O-acetyl-)-ß-D-glycoside protected LPS-induced macrophage RAW 264.7 cells from injury. The flavone glycoside markedly inhibited the LPS-induced production of tumor necrosis factor-α, interleukin-1ß, and interleukin-6 and increased interleukin-10 release in a concentration-dependent manner. Furthermore, treatment with the flavone glycoside decreased nitric oxide and prostaglandin E2 in LPS-challenged RAW 264.7 cells. These decreases were associated with the down-regulation of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), and nuclear factor kappa B (NF-κB) activity. These findings suggest that the anti-inflammatory effects of selagin-7-O-(6''-O-acetyl-)-ß-D-glycoside were associated with the adjustment of inflammatory cytokines, and attributed to the down-regulation of NF-κB and consequent suppression of the expression of iNOS and COX-2.