ABSTRACT
Background: Gegen Qinlian decoction (GGQLD) is a typical traditional Chinese medicine (TCM) prescription documented in Shang Han Lun. Clinically, GGQLD has been utilized to manage the inflammatory symptoms of metabolic diseases and to protect against renal damage in China. In the present study, a hypothesis was proposed that the multi-target solution of GGQLD produced anti-inflammatory effects on ameliorating hyperuricemia (HUA). Methods: A total of 30 primary HUA patients receiving GGQLD treatment (two doses daily) for 4 weeks were selected. Then, differences in uric acid (UA) levels and expression of peripheral blood mononuclear cells (PBMCs) and urinary exosomes before and after treatment were analyzed. The therapeutic indexes for the active ingredients in GGQLD against HUA were confirmed through pharmacological subnetwork analysis. Besides, the HUA rat model was established through oral gavage of potassium oxonate and treated with oral GGQLD. In addition, proximal tubular epithelial cells (PTECs) were stimulated by UA and intervened with GGQLD for 48 h. Subsequently, RNA-seq, flow cytometry, and confocal immunofluorescence microscopy were further conducted to characterize the differences in UA-mediated inflammation and apoptosis of human renal tubular epithelial cells pre- and post-administration of GGQLD. In the meanwhile, quantitative real-time PCR (qPCR) was carried out to determine gene expression, whereas a western blotting (WB) assay was conducted to measure protein expression. Results: Our network analysis revealed that GGQLD treated HUA via the anti-inflammatory and antiapoptotic pathways. Additionally, NLPR3 expression significantly decreased in PBMCs and urinary exosomes of HUA patients after GGQLD treatment. In vivo, GGQLD treatment alleviated HUA-induced renal inflammation, which was associated with decreased expression of NLRP3 inflammasomes and apoptosis-related mRNAs. Moreover, GGQLD promoted renal UA excretion by inhibiting the activation of GSDMD-dependent pyroptosis induced by NLRP3 inflammasomes and by reducing apoptosis via the mitochondrial apoptosis signaling pathway in vitro. Conclusion: This study indicates that GGQLD efficiently reduces inflammatory responses while promoting UA excretion in HUA. Our findings also provide compelling evidence supporting the idea that GGQLD protects against the UA-mediated renal tubular epithelial cell inflammation through the mitochondrial apoptosis signaling pathways. Taken together, these findings have demonstrated a novel therapeutic method for the treatment of HUA.
ABSTRACT
Microbial fermentation is a promising technology for hydrogen (H(2)) production. H(2) producers in marine geothermal environments are thermophilic and halotolerant. However, no one has surveyed an environment specifically for thermophilic bacteria that produce H(2) through Fe-Fe hydrogenases (H(2)ase). Using heterotrophic medium, several microflora from a seaweed bed associated with marine hot springs were enriched and analyzed for H(2) production. A H(2)-producing microflora was obtained from Sargassum sp., 16S rRNA genes and Fe-Fe H(2)ase diversities of this enrichment were also analyzed. Based on 16S rRNA genes analysis, 10 phylotypes were found in the H(2)-producing microflora showing 90.0-99.5 % identities to known species, and belonged to Clostridia, Gammaproteobacteria, and Bacillales. Clostridia were the most abundant group, and three Clostridia phylotypes were most related to known H(2) producers such as Anaerovorax odorimutans (94.0 % identity), Clostridium papyrosolvens (98.4 % identity), and Clostridium tepidiprofundi (93.1 % identity). For Fe-Fe H(2)ases, seven phylotypes were obtained, showing 63-97 % identities to known Fe-Fe H(2)ases, and fell into four distinct clusters. Phylotypes HW55-3 and HM55-1 belonged to thermophilic and salt-tolerant H(2)-producing Clostridia, Halothermothrix orenii-like Fe-Fe H(2)ases (80 % identity), and cellulolytic H(2)-producing Clostridia, C. papyrosolvens-like Fe-Fe H(2)ases (97 % identity), respectively. The results of both 16S rRNA genes and Fe-Fe H(2)ases surveys suggested that the thermophilic and halotolerant H(2)-producing microflora in seaweed bed of hot spring area represented previously unknown H(2) producers, and have potential application for H(2) production.
Subject(s)
Hot Springs/microbiology , Hydrogen/metabolism , Hydrogenase/metabolism , Iron-Sulfur Proteins/metabolism , Seaweed/metabolism , Seaweed/microbiology , Bacillales/classification , Bacillales/genetics , Clostridium/classification , Clostridium/genetics , Gammaproteobacteria/classification , Gammaproteobacteria/genetics , Hydrogenase/genetics , Indonesia , Iron-Sulfur Proteins/genetics , Metagenome , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/geneticsABSTRACT
Calcium-activated potassium channels on plasma membrane enable potassium influx into the cell with ensuing changes in plasma membrane potential and consequent effects on cellular metabolic functions. Recently, this potassium channel was reported to regulate the cellular responses of mammalian immune cells. We have postulated the presence of such a channel in fish immune cells and its potential role in immunoregulation in fish. Employing specific primers and RNA template, we cloned a segment of a novel gene from turbot blood sample and subsequently obtained a full cDNA sequence using RACE approaches. Bioinformatic analysis revealed structural and phylogenetic characteristics of a novel small conductance calcium-activated potassium channel gene, we called TSKCa, which exhibits homologous domains to other species particularly in the transmembrane regions. Full-length TSKCa cDNA is 1698 bp with a 1632 bp open reading frame encoding a protein of 544 amino acids. TSKCa gene is expressed in majority of the tested organs and tissues of turbot. To assess the postulated immune function of TSKCa, we infected turbot with the pathogen Vibrio anguillarum. Here, semi-quantitative RT-PCR analysis demonstrated increased mRNA expression of TSKCa in head kidney, spleen and blood, indicating an important role of TSKCa in these organ tissues that mediate the immune defense response of turbot. In contrast, there was less change in expression in the turbot intestines and liver which were less implicated in the immune response in present study.
Subject(s)
Flatfishes/genetics , Flatfishes/metabolism , Gene Expression Regulation , Small-Conductance Calcium-Activated Potassium Channels/genetics , Small-Conductance Calcium-Activated Potassium Channels/metabolism , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Fish Diseases/immunology , Gene Expression Profiling , Molecular Sequence Data , Phylogeny , RNA, Messenger/metabolism , Sequence Alignment , Vibrio/physiology , Vibrio Infections/immunologyABSTRACT
Volatiles emitted from the leaves of Lycopersicon esculentum at the two-, ten-leaf and anthesis periods were collected by a gas absorbing method and analyzed by gas chromatography (GC)-mass spectrometry. In total, 33 compounds of volatiles emitted from three developmental stage plants were separated and identified, and quantitatively analyzed by the internal standard addition method. All of the samples of volatile were found to be rich in monoterpenes and sesquiterpenes. beta-phellandrene and caryophyllene predominated in the volatiles of the leaves of plants at the two- and ten-leaf stages. Furthermore, (E)-2-hexenal were the dominant components in the volatiles emitted from anthesis plants. The results of volatiles analyzed show that the compositions varied depending on the developmental stages. The volatiles emitted from crushed tomato leaves of plants at the anthesis stage had the most strongly inhibitory activity against the spore germination and hyphal growth of Botrytis cinerea and Fusarium oxysporum, followed by ten- and two-leaf plants. However, the activity of volatiles, emitted from the leaves of plants at the two-leaf stage, in inhibiting F. oxysporum was greater than B. cinerea.
Subject(s)
Botrytis/drug effects , Fusarium/drug effects , Plant Exudates/pharmacology , Plant Leaves/metabolism , Solanum lycopersicum/metabolism , Botrytis/physiology , Cyclohexane Monoterpenes , Cyclohexenes/chemistry , Cyclohexenes/metabolism , Cyclohexenes/pharmacology , Fusarium/physiology , Gas Chromatography-Mass Spectrometry , Hexobarbital/chemistry , Hexobarbital/metabolism , Hexobarbital/pharmacology , Hyphae/drug effects , Hyphae/growth & development , Monoterpenes/chemistry , Monoterpenes/metabolism , Monoterpenes/pharmacology , Plant Exudates/chemistry , Plant Exudates/metabolism , Polycyclic Sesquiterpenes , Sesquiterpenes/chemistry , Sesquiterpenes/metabolism , Sesquiterpenes/pharmacology , Spores, Fungal/drug effects , Spores, Fungal/physiology , VolatilizationABSTRACT
Wheat (Triticum aestivum L.) seedlings were grown in taurine solution at concentrations of 10, 100, 500, 1000 and 5000 mg/L, and the photochemistry efficiency, the relative permeability of membrane, membrane lipid peroxidation and the growth indexes in wheat seedlings were determined. The results showed that taurine treatments distinctly promoted the growth of wheat seedlings and increased root length, plant height, dry weight and fresh weight single plant of wheat seedlings. In addition, the photochemistry efficiency in the leaf was also increase, and the membrane relative permeability and the content of membrane lipid peroxidation product MDA were decreased, with an optimum treatment concentration at about 500 mg/L. It suggests that taurine has protective effects on the cell membrane of wheat seedlings to the certain extent.