The "time-response" effect of Wenyang Pingchuan Formula on miR-19a in asthmatic mice experiment.

BACKGROUND: Wenyang Pingchuan Formula (WPCF) is an empirical formula for the treatment of acute childhood asthma. However, the "time-effect" relationship of this prescription is not clear. This paper explores the relationship between Janus activated kinase signal transducer and activator of transcriptions (JAK/STAT), nuclear factor-κB (NF-κB), and microRNA (miR-19a), and also preliminarily determines the best time-effect relationship of WPCF in reducing the airway inflammation in asthmatic mice. METHOD: 80 BALB/c mice were randomly divided into four groups: control (CON) group, model (MDL) group, dexamethasone (DEX) group, and WPCF group. MDL group was established through intraperitoneal injection of 10% ovalbumin (OVA) and Al(OH)3 solution and the inhalation of aerosolized 5% OVA solution. Enzyme-linked immunosorbent assay (ELISA), real-time PCR and Western blot were conducted to determine the levels of interleukin (IL)-4, IL-13, interferon-γ (IFN-γ) in bronchoalveolar lavage fluid (BALF), contents of miR-19a mRNA and STAT6, phosphorylated signal transducers and activators of transcription 6 (p-STAT6), p65, phosphorylated p65 (p-p65), suppressors of cytokine signaling 1 (SOCS1), and tumor necrosis factor α-induced protein-3 (Tnfaip3) proteins after 7 and 28 days of intervention respectively. RESULTS: Significant down-regulation of IL-4 and IL-13 expression (P<0.05) and up-regulation of IFN-γ expression (P<0.05) in BALF have been observed for WPCF group compared with the MDL group. The significant down-regulation of miR-19a mRNA and STAT6, p-STAT6, p65, p-p65 proteins (P<0.05) and up-regulation of SOCS1 and Tnfaip3 proteins (P<0.05) in BALF was also observed for WPCF group compared to the MDL group. During the experiment, the weight of the mice in DEX group significantly decreased (P<0.05) compared with the other groups. CONCLUSIONS: WPCF could restore Th1/Th2 balance. The longer the intervention time, the more effective the treatment. The down-regulation of miR-19a mRNA by activating JAK/STAT and NF-κB signal pathways may be a possible mechanism by which WPCF alleviates airway inflammation.

Pingchuan formula improves allergic asthma in mice through inhibiting nuclear factor-kappa B / mitogen-activated protein kinase signaling pathway.

OBJECTIVE: To examine the role and decipher the mechanism of Pingchuan formula (PCF) in treating allergic asthma. METHODS: The mice were treated with saline, dexamethasone (DXM) and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance (RI), pulmonary dynamic compliance (Cdyn) and maximum voluntary ventilation (MVV) were measured. In addition, cellular changes in bronchoalveolar lavage fluid (BALF) and pathological changes in lung biopsy as well as the expression level of -smooth muscle actin (α-SMA), transforming growth factor-beta1 (TGF-α1) in BALF and interleukin-5 (IL-5), interleukin-13 (IL-13), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), nuclear factor-kappa B-p65 (NF-κBp65), inhibitor-α of nuclear transcription factor κB (IκBα), p38 mitogen-activated protein kinase (p38MAPK), c-jun n-terminal kinase (JNK) and its phosphorylated proteins in lung tissue were also examined and compared among different groups. RESULTS: Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group. Both DXM and PCF effectively decreased the number of eosinophils, lymphocytes, and neutrophils in BAL as well as the secretion of α-SMA and TGF-α1, IL-5, IL-13, while increased the expression of TNF-α and IFN-γ. Furthermore, our study indicated that the NF-κBp65, IκBα, p38MAPK and JNK pathways were inhibited under the treatment of PCF. CONCLUSION: Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway. This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of thediseasepathogenesis.

Pingchuan formula attenuates airway mucus hypersecretion via regulation of the PNEC-GABA-IL13-Muc5ac axis in asthmatic mice.

BACKGROUND/AIMS: Asthma is a common chronic respiratory disease. It has been reported that Pingchuan formula (PCF) can control asthma attacks by reducing airway inflammation, muscle spasm and mucus secretion. However, PCF's mechanism for reducing airway mucus hypersecretion remains unclear. This study aimed to investigate the effect of PCF on airway mucus secretion in asthmatic mice and to explore changes in the PNEC-GABA-IL13-Muc5ac axis. METHODS: Male Babl/c mice were used to establish the asthma model via sensitisation with OVA. Mice were randomly divided into Normal, OVA, DEX, and PCF groups. After treatment, lung histopathology was observed with H&E and PAS staining. BALF levels of IL-5 and IL-13 were detected using ELISA. The levels of mRNA and protein expression for GAD1, GABAARß1, GABAARα1 and Muc5ac in the lung tissue were measured by RT-PCR and Western blot assays. PNECs were observed with AgNOR staining. RESULTS: PCF treatment effectively reduced goblet cell (P < 0.01) and PNEC (P < 0.05) proliferation, lung tissue inflammation and airway mucus hypersecretion. In addition, PCF also markedly downregulated mRNA and protein expression of GAD1, GABAARß1, GABAARα1 and Muc5ac (P < 0.05, compared with OVA), thus inhibiting the GABA-IL-13 pathway in the lung tissue of asthmatic mice. CONCLUSION: These findings suggest that PCF controls asthma attacks by reducing airway inflammation and mucus hypersecretion via the PNEC-GABA-IL13-Muc5ac axis.

Genome-wide identification and evolution of TC1/Mariner in the silkworm (Bombyx mori) genome.

TC1/Mariner transposons belong to class II transposable elements (TEs) that use DNA-mediated "cut and paste" mechanism to transpose, and they have been identified in almost all organisms. Although silkworm (Bombyx mori) has a large amount of TC1/Mariner elements, the genome wide information of this superfamily in the silkworm is unknown. In this study, we have identified 2670 TC1/Mariner (Bmmar) elements in the silkworm genome. All the TEs were classified into 22 families by means of fgclust, a tool of repetitive sequence classification, seven of which was first reported in this study. Phylogenetic and structure analyses based on the catalytic domain (DDxD/E) of transposase sequences indicated that all members of TC1/Mariner were grouped into five subgroups: Mariner, Tc1, maT, DD40D and DD41D/E. Of these five subgroups, maT rather than Mariner possessed most members of TC1/Mariner (51.23%) in the silkworm genome. In particular, phylogenetic analysis and structure analysis revealed that Bmmar15 (DD40D) formed a new basal subgroup of TC1/Mariner element in insects, which was referred to as bmori. Furthermore, we concluded that DD40D appeared to intermediate between mariner and Tc1. Finally, we estimated the insertion time for each copy of TC1/Mariner in the silkworm and found that most of members were dramatically amplified during a period from 0 to 1 mya. Moreover, the detailed functional data analysis showed that Bmmar1, Bmmar6 and Bmmar9 had EST evidence and intact transposases. These implied that TC1/Mariner might have potential transpositional activity. In conclusion, this study provides some new insights into the landscape, origin and evolution of TC1/Mariner in the insect genomes.

Unexpected invasion of miniature inverted-repeat transposable elements in viral genomes.

BACKGROUND: Transposable elements (TEs) are common and often present with high copy numbers in cellular genomes. Unlike in cellular organisms, TEs were previously thought to be either rare or absent in viruses. Almost all reported TEs display only one or two copies per viral genome. In addition, the discovery of pandoraviruses with genomes up to 2.5-Mb emphasizes the need for biologists to rethink the fundamental nature of the relationship between viruses and cellular life. RESULTS: Herein, we performed the first comprehensive analysis of miniature inverted-repeat transposable elements (MITEs) in the 5170 viral genomes for which sequences are currently available. Four hundred and fifty one copies of ten miniature inverted-repeat transposable elements (MITEs) were found and each MITE had reached relatively large copy numbers (some up to 90) in viruses. Eight MITEs belonging to two DNA superfamilies (hobo/Activator/Tam3 and Chapaev-Mirage-CACTA) were for the first time identified in viruses, further expanding the organismal range of these two superfamilies. TEs may play important roles in shaping the evolution of pandoravirus genomes, which were here found to be very rich in MITEs. We also show that putative autonomous partners of seven MITEs are present in the genomes of viral hosts, suggesting that viruses may borrow the transpositional machinery of their cellular hosts' autonomous elements to spread MITEs and colonize their own genomes. The presence of seven similar MITEs in viral hosts, suggesting horizontal transfers (HTs) as the major mechanism for MITEs propagation. CONCLUSIONS: Our discovery highlights that TEs contribute to shape genome evolution of pandoraviruses. We concluded that as for cellular organisms, TEs are part of the pandoraviruses' diverse mobilome.

The Antioxidant Inhibition of Clove Effective Fraction on Lipid, Protein and Spectra Variation of LDL.

Oxidation of low density lipoprotein (LDL) has been considered as the critical factor which led to atherosclerosis (AS). Lipid and protein in LDL were oxidized to cause change of spectra during oxidation. Clove has been demonstrated to possess the strongest antioxidant capacity among 87 both medical and edible plants proclaimed by China. However, whether LDL oxidation is inhibited by clove? If so, whether it inhibits the oxidation of lipid and protein in LDL, and how does spectral characteristic of LDL change during oxidation when clove was added into LDL. Currently, these questions are still unclear. Therefore, the inhibition efficiency of the effective fraction from clove (EFC) on oxidation of LDL was studied by UV-Visible spectra and fluorescence spectra. The results indicated that EFC might effectively delay propagation of conjugated diene (CD) during LDL oxidation and postpone reaching maximum of its content. Likewise, it might inhibit cholesterol degradation, tryptophan (Trp) fluorescence quenching, lysine (Lys) residues from being modified, and lipofuscins from being generated in peroxidatic reaction among lipid and protein. Besides, EFC also might affect variation of UV-Visible spectra of LDL during oxidation. This study provides reference for future research and development of clove functional food inhibiting AS.