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Bacterial small molecule metabolites such as adenosine-diphosphate-d-glycero-ß-d-manno-heptose (ADP-heptose) and their derivatives act as effective innate immune agonists in mammals. We show that functional nucleotide-diphosphate-heptose biosynthetic enzymes (HBEs) are distributed widely in bacteria, archaea, eukaryotes, and viruses. We identified a conserved STTR5 motif as a hallmark of heptose nucleotidyltransferases that can synthesize not only ADP-heptose but also cytidine-diphosphate (CDP)- and uridine-diphosphate (UDP)-heptose. Both CDP- and UDP-heptoses are agonists that trigger stronger alpha-protein kinase 1 (ALPK1)-dependent immune responses than ADP-heptose in human and mouse cells and mice. We also produced ADP-heptose in archaea and verified its innate immune agonist functions. Hence, the ß-d-manno-heptoses are cross-kingdom, small-molecule, pathogen-associated molecular patterns that activate the ALPK1-dependent innate immune signaling cascade.
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Heptosas , Nucleotidiltransferasas , Moléculas de Patrón Molecular Asociado a Patógenos , Animales , Humanos , Ratones , Secuencias de Aminoácidos , Archaea/enzimología , Bacterias/enzimología , Bacterias/metabolismo , Heptosas/biosíntesis , Heptosas/inmunología , Inmunidad Innata , Nucleotidiltransferasas/química , Nucleotidiltransferasas/clasificación , Nucleotidiltransferasas/genética , Moléculas de Patrón Molecular Asociado a Patógenos/inmunología , Moléculas de Patrón Molecular Asociado a Patógenos/metabolismo , Proteínas Quinasas/metabolismo , Virus/enzimologíaRESUMEN
Cytotoxic lymphocyte-derived granzyme A (GZMA) cleaves GSDMB, a gasdermin-family pore-forming protein1,2, to trigger target cell pyroptosis3. GSDMB and the charter gasdermin family member GSDMD4,5 have been inconsistently reported to be degraded by the Shigella flexneri ubiquitin-ligase virulence factor IpaH7.8 (refs. 6,7). Whether and how IpaH7.8 targets both gasdermins is undefined, and the pyroptosis function of GSDMB has even been questioned recently6,8. Here we report the crystal structure of the IpaH7.8-GSDMB complex, which shows how IpaH7.8 recognizes the GSDMB pore-forming domain. We clarify that IpaH7.8 targets human (but not mouse) GSDMD through a similar mechanism. The structure of full-length GSDMB suggests stronger autoinhibition than in other gasdermins9,10. GSDMB has multiple splicing isoforms that are equally targeted by IpaH7.8 but exhibit contrasting pyroptotic activities. Presence of exon 6 in the isoforms dictates the pore-forming, pyroptotic activity in GSDMB. We determine the cryo-electron microscopy structure of the 27-fold-symmetric GSDMB pore and depict conformational changes that drive pore formation. The structure uncovers an essential role for exon-6-derived elements in pore assembly, explaining pyroptosis deficiency in the non-canonical splicing isoform used in recent studies6,8. Different cancer cell lines have markedly different isoform compositions, correlating with the onset and extent of pyroptosis following GZMA stimulation. Our study illustrates fine regulation of GSDMB pore-forming activity by pathogenic bacteria and mRNA splicing and defines the underlying structural mechanisms.
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Gasderminas , Proteínas Citotóxicas Formadoras de Poros , Animales , Humanos , Ratones , Línea Celular Tumoral , Microscopía por Crioelectrón , Cristalografía por Rayos X , Gasderminas/química , Gasderminas/genética , Gasderminas/metabolismo , Gasderminas/ultraestructura , Proteínas de Neoplasias/química , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Proteínas de Neoplasias/ultraestructura , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Proteínas Citotóxicas Formadoras de Poros/ultraestructura , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Isoformas de Proteínas/ultraestructura , Piroptosis , Shigella flexneri , Especificidad de la Especie , Empalme AlternativoRESUMEN
Bacterial biofilm widely exists in all kinds of bacteria, and is related to about 80 percent of bacterial infections. It is one of the main reasons for bacterial tolerance and resistance to existing antibiotics. Therefore, there is unmet clinical need for new anti-biofilm drugs. At present, there are three kinds of anti-biofilm agents under research, including biofilm inhibitors, biofilm dispersal agents and biofilm eradication agents. Among them, the biofilm eradication agent is unique, which can not only kill bacteria in biofilm but also eliminate biofilm as a monotherapy. Based on modifications of natural products with antibacterial activity, a variety of compounds with biofilm eradicating activities have been obtained, such as, acyldepsipeptides, pyrrolomycins, halogenated phenazines and halogenated 8-hydroxyquinolines. In this review, we summarize several major biofilm eradication agents above according to their structures and mechanisms.
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The circadian rhythm of mammals is a physiological phenomena that is about 24 hours produced by genetically encoded molecular clocks, making the physiological process of the body coordinated with the changes of the external environment, and it is a manifestation of adaptation to the environment. In mammals, reproductive physiology is regulated by the circadian clock. The expression of circadian clock genes has been observed in each tissue of the hypothalamic-pituitary-ovarian (HPO) axis, and the biological clock at all levels coordinates and synchronizes with each other to maintain normal reproductive behavior. The production, maintenance, and regulation of circadian rhythms depend on a chain of transcription-translation feedback loops (TTLs), which determine the cycle and amplitude of gene expression in each tissue of the HPO axis. The circadian clock of the ovary is regulated by theneuroendocrine regulation of suprachiasmatic nucleus of the hypothalamus, but it is autonomous. Circadian rhythm disruption caused by environmental factors can seriously impair female fertility and lead to a range of related ovarian diseases. In addition, the circadian clock is also closely related to ovarianaging. Based on existing research, this paper focuses on the mechanism of the circadian clock in ovarian follicular development, ovulation and steroid generation, as well as the latest research progress on the relationship between the circadian clock and ovarian aging. In addition, several common ovarian diseases with decreased fertility due to circadian clock disorders are described.
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Besides UCP1-dependent thermogenesis pathways, UCP1-independent thermogenesis pathways also could increase heat production in adipose tissue to combat obesity. N-Acyl amino acids (NAAs) have been suggested as novel endogenous uncouplers to induce mitochondria UCP1-independent thermogenesis in adipose tissue. Here, we use mouse skeletal muscle C2C12 cells which lack of UCP1 as UCP1 negative cell models. Comparing with its corresponding common fatty acid—oleate, one of the NAAs—N-Oleoylglycine (NOGly), which is highly expressed in the plasma of HFD mice, is selected to study their effects and mechanisms on mitochondrial thermogenesis. We found that 60 μmol / L oleate could induce mitochondrial oxidative phosphorylation protein levels, as well as increase mitochondria thermogenesis-related genes (COX8b, DIO2, UCP3) expression (P < 0. 05) . However, 60 μmol / L NOGly damaged the production and oxidative phosphorylation of mitochondria, significantly down-regulated expression of thermogenic genes (PGC1a, COX8b, COX2, DIO2, UQCRFS1and UCP3) (P< 0. 01), induced the production of reactive oxygen species (ROS) in the mitochondria, and enhanced the oxidative stress in cells. Our study found that oleate can induce UCP1-independent thermogenesis under 60 μmol / L addition dose, whereas NOGly does not due to the induction of oxidative stress in cells.
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Oocytes are the germ cells of female animals, which determine the reproductive ability of female animals. A large amount of lipids are present in oocytes, which are found in lipid droplets mostly in the form of triglycerides. The size, color and distribution pattern of lipid droplets are associated with the developmental ability of oocytes. Triglycerides could be lipolyzed into fatty acids in oocytes. The fatty acid β-oxidation is an important energy source for the development of oocytes and early embryos. However, excessive lipid deposition would increase levels of reactive oxygen species (ROS), resulting in the dysfunction of mitochondria and endoplasmic reticulum, eventually impairing the subsequent oocyte development. By summarizing the positive and negative effects of lipids on oocyte development, this review shows the dual roles of lipids in oocyte development, and discusses the effects of lipids on oocyte development.
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Echinenone and canthaxanthin are important carotenoid pigments with food and industrial applications. Biosynthesis of echinenone and/or canthaxanthin is catalyzed by ß-carotene ketolase (CrtO), with ß-carotene as the substrate. In this study, we generated transgenic Nostoc sp. PCC 7120 overexpressing a heterologous crtO gene from Nostoc flagelliforme and evaluated the productivity of both pigments. Normal (BG11 medium, 30⯰C) and osmotic stress (BG11 medium supplemented with 0.4â¯M mannitol, 30⯰C) conditions were used for cultivation. As compared to control strain, production of echinenone and canthaxanthin in transgenic strain were respectively increased by more than 16 % and 80 %, under either normal or osmotic stress conditions. Especially upon the stress condition, higher proportion of echinenone and canthaxanthin in total pigments was achieved, which should be beneficial for downstream separation and purification. In addition, transgenic strain showed drought tolerance and could revive from desiccation treatment after rewetting. Thus, this study provided technical clues for production of both pigments in engineered cyanobacteria as well as for cyanobacterial anhydrobiotic engineering.
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Nostoc/genética , Organismos Modificados Genéticamente/crecimiento & desarrollo , Organismos Modificados Genéticamente/metabolismo , Oxigenasas/genética , Adaptación Fisiológica , Proteínas Bacterianas/genética , Cantaxantina/biosíntesis , Carotenoides/metabolismo , Clonación Molecular , Sequías , Genes Bacterianos , Ingeniería Metabólica/métodos , Nostoc/crecimiento & desarrollo , Nostoc/metabolismo , Organismos Modificados Genéticamente/genética , Oxigenasas/metabolismo , beta Caroteno/biosíntesisRESUMEN
BACKGROUND: Periodontitis was reported to be associated with inflammatory bowel disease (IBD). However, the association between them has not been firmly established in the existing literature. Therefore, this meta-analysis was conducted to evaluate the relationship between periodontitis and IBD. METHODS: Electronic databases were searched for publications up to August 1, 2019 to include all eligible studies. The pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were estimated to determine the association between periodontal disease and IBD using a random or fixed effects model according to heterogeneity. RESULTS: Six eligible studies involving 599 IBD patients and 448 controls were included. The pooled OR between periodontitis and IBD was 3.17 (95% CI: 2.09-4.8) with no heterogeneity observed (I2 = 0.00%). The pooled ORs were 3.64 (95% CI: 2.33-5.67) and 5.37 (95% CI: 3.30-8.74) for the associations between periodontitis and the two sub-categories of IBD, Crohn' s disease and ulcerative colitis, respectively. CONCLUSIONS: The results demonstrated that periodontitis was significantly associated with IBD. However, the mechanisms underlying periodontitis and IBD development are undetermined. Further studies are needed to elucidate this relationship.
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Colitis Ulcerosa/epidemiología , Enfermedad de Crohn/epidemiología , Enfermedades Inflamatorias del Intestino/epidemiología , Periodontitis/epidemiología , Adulto , Humanos , Persona de Mediana Edad , Higiene Bucal , Prevalencia , Factor de Necrosis Tumoral alfa , Adulto JovenRESUMEN
Chemical cross-linking of proteins coupled with mass spectrometry analysis (CXMS) is widely used to study protein-protein interactions (PPI), protein structures, and even protein dynamics. However, structural information provided by CXMS is still limited, partly because most CXMS experiments use lysine-lysine (K-K) cross-linkers. Although superb in selectivity and reactivity, they are ineffective for lysine deficient regions. Herein, we develop aromatic glyoxal cross-linkers (ArGOs) for arginine-arginine (R-R) cross-linking and the lysine-arginine (K-R) cross-linker KArGO. The R-R or K-R cross-links generated by ArGO or KArGO fit well with protein crystal structures and provide information not attainable by K-K cross-links. KArGO, in particular, is highly valuable for CXMS, with robust performance on a variety of samples including a kinase and two multi-protein complexes. In the case of the CNGP complex, KArGO cross-links covered as much of the PPI interface as R-R and K-K cross-links combined and improved the accuracy of Rosetta docking substantially.
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Arginina/química , Reactivos de Enlaces Cruzados/química , Lisina/química , Espectrometría de Masas/métodos , Proteínas/química , Algoritmos , Arginina/metabolismo , Lisina/metabolismo , Modelos Moleculares , Estructura Molecular , Péptidos/química , Péptidos/metabolismo , Conformación Proteica , Mapas de Interacción de Proteínas , Proteínas/metabolismoRESUMEN
BACKGROUND: The protein encoded by the selenoprotein S gene is considered to be an anti-inflammatory and antioxidant protein and is involved in a variety of diseases. Therefore, we want to study the distribution characteristics of this gene in Chinese diabetic population. METHODS: A total of 170 patients with DM (including 100 patients with T2DM and 70 patients with diabetic nephropathy [DN]) and 100 healthy controls (HC) were selected from Haikou People's Hospital (China) between January 2017 and July 2017. The polymorphisms of three SEPS1 genes (SNP ID: rs4965814, rs28665122, and rs34713741) were measured by massARRAY method, while the polymorphisms of SEPS1 genes (SNP ID: rs4965373) were detected by Sanger sequencing. RESULTS: Comparing three groups, the results were the following: (a) There was a significant difference in the genotype and allele distribution of rs34713741 between DN group and HC group and between T2DM group and DN group; For this gene locus, the risk of diabetic nephropathy in healthy individuals with T allele was 0.6 times higher than that in individuals with GG genotype (OR = 0.60, 95% CI: 0.46 ~ 0.77). (b) There was a significant difference in the distribution of rs4975814 genotype between DN group and HC group; for this gene locus, the risk of diabetic nephropathy in healthy individuals with T allele was 2.71 times higher than that in individuals with GG genotype (OR = 2.71, 95% CI: 1.66 ~ 4.45). CONCLUSION: We conclude that rs34713741 (GT + TT) may be a protective gene for DN and the rs4975814 (GT + TT) may be a susceptibility gene for DN.
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Pueblo Asiatico/genética , Diabetes Mellitus Tipo 2/complicaciones , Nefropatías Diabéticas/diagnóstico , Predisposición Genética a la Enfermedad , Proteínas de la Membrana/genética , Polimorfismo de Nucleótido Simple , Selenoproteínas/genética , Adulto , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2/genética , Nefropatías Diabéticas/etiología , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Factores de RiesgoRESUMEN
The present study aimed to investigate the expression of pyrroline-5-carboxylate reductase 1 (P5CR1) protein in lung adenocarcinoma and paracancerous tissues and to explore the effect of silencing the encoding gene PYCR1 on the proliferation, migration, invasion, and cisplatin sensitivity in lung adenocarcinoma cells, thereby providing a novel therapeutic target for the treatment of the disease. Immunohistochemistry staining was used to detect the P5CR1 protein expression in lung adenocarcinoma and paracancerous tissues, and statistical analysis evaluated the correlation between P5CR1 protein expression and gender, age, tissue part, or pathological grade. The CCK8 assay was performed to detect the proliferation and cisplatin sensitivity, while the effect of PYCR1 on the migration and invasion of lung adenocarcinoma cells was detected by scratch test and transwell chamber assay. The findings demonstrated that the P5CR1 protein expression was significantly elevated in lung adenocarcinoma tissues and correlated with the pathological grade, whereas no significant correlation was established between the protein expression and gender, age, or tissue part. Furthermore, after PYCR1 gene silencing, the proliferation and invasion were significantly suppressed, while the sensitivity to cisplatin was significantly enhanced. Therefore, it can be speculated that the PYCR1 gene affects the biological behavior of lung adenocarcinoma and cisplatin resistance, serving as a potential therapeutic target for lung adenocarcinoma.
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Immune recognition of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors often activates proinflammatory NF-κB signalling1. Recent studies indicate that the bacterial metabolite D-glycero-ß-D-manno-heptose 1,7-bisphosphate (HBP) can activate NF-κB signalling in host cytosol2-4, but it is unclear whether HBP is a genuine PAMP and the cognate pattern recognition receptor has not been identified. Here we combined a transposon screen in Yersinia pseudotuberculosis with biochemical analyses and identified ADP-ß-D-manno-heptose (ADP-Hep), which mediates type III secretion system-dependent NF-κB activation and cytokine expression. ADP-Hep, but not other heptose metabolites, could enter host cytosol to activate NF-κB. A CRISPR-Cas9 screen showed that activation of NF-κB by ADP-Hep involves an ALPK1 (alpha-kinase 1)-TIFA (TRAF-interacting protein with forkhead-associated domain) axis. ADP-Hep directly binds the N-terminal domain of ALPK1, stimulating its kinase domain to phosphorylate and activate TIFA. The crystal structure of the N-terminal domain of ALPK1 and ADP-Hep in complex revealed the atomic mechanism of this ligand-receptor recognition process. HBP was transformed by host adenylyltransferases into ADP-heptose 7-P, which could activate ALPK1 to a lesser extent than ADP-Hep. ADP-Hep (but not HBP) alone or during bacterial infection induced Alpk1-dependent inflammation in mice. Our findings identify ALPK1 and ADP-Hep as a pattern recognition receptor and an effective immunomodulator, respectively.
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Azúcares de Adenosina Difosfato/inmunología , Burkholderia cenocepacia , Citosol , Inmunidad Innata , Moléculas de Patrón Molecular Asociado a Patógenos/inmunología , Proteínas Quinasas/metabolismo , Yersinia pseudotuberculosis , Azúcares de Adenosina Difosfato/metabolismo , Animales , Infecciones por Burkholderia/enzimología , Infecciones por Burkholderia/inmunología , Infecciones por Burkholderia/patología , Burkholderia cenocepacia/genética , Burkholderia cenocepacia/inmunología , Burkholderia cenocepacia/metabolismo , Sistemas CRISPR-Cas , Cristalografía por Rayos X , Citocinas/biosíntesis , Citosol/enzimología , Citosol/inmunología , Disacáridos/metabolismo , Activación Enzimática , Femenino , Edición Génica , Factores Inmunológicos/inmunología , Factores Inmunológicos/metabolismo , Inmunomodulación , Inflamación/enzimología , Inflamación/inmunología , Inflamación/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Moleculares , FN-kappa B/metabolismo , Moléculas de Patrón Molecular Asociado a Patógenos/metabolismo , Yersinia pseudotuberculosis/genética , Yersinia pseudotuberculosis/inmunología , Yersinia pseudotuberculosis/metabolismoRESUMEN
Inflammatory caspases cleave the gasdermin D (GSDMD) protein to trigger pyroptosis, a lytic form of cell death that is crucial for immune defences and diseases. GSDMD contains a functionally important gasdermin-N domain that is shared in the gasdermin family. The functional mechanism of action of gasdermin proteins is unknown. Here we show that the gasdermin-N domains of the gasdermin proteins GSDMD, GSDMA3 and GSDMA can bind membrane lipids, phosphoinositides and cardiolipin, and exhibit membrane-disrupting cytotoxicity in mammalian cells and artificially transformed bacteria. Gasdermin-N moved to the plasma membrane during pyroptosis. Purified gasdermin-N efficiently lysed phosphoinositide/cardiolipin-containing liposomes and formed pores on membranes made of artificial or natural phospholipid mixtures. Most gasdermin pores had an inner diameter of 1014 nm and contained 16 symmetric protomers. The crystal structure of GSDMA3 showed an autoinhibited two-domain architecture that is conserved in the gasdermin family. Structure-guided mutagenesis demonstrated that the liposome-leakage and pore-forming activities of the gasdermin-N domain are required for pyroptosis. These findings reveal the mechanism for pyroptosis and provide insights into the roles of the gasdermin family in necrosis, immunity and diseases.
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Permeabilidad de la Membrana Celular/efectos de los fármacos , Proteínas/antagonistas & inhibidores , Proteínas/química , Piroptosis , Animales , Biomarcadores de Tumor/antagonistas & inhibidores , Biomarcadores de Tumor/química , Biomarcadores de Tumor/metabolismo , Cardiolipinas/metabolismo , Caspasas/metabolismo , Línea Celular , Membrana Celular/química , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Cristalografía por Rayos X , Proteínas de Unión al ADN/antagonistas & inhibidores , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular , Liposomas , Lípidos de la Membrana/metabolismo , Ratones , Modelos Moleculares , Necrosis , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Proteínas de Unión a Fosfato , Fosfatidilinositoles/metabolismo , Porosidad/efectos de los fármacos , Estructura Terciaria de Proteína , Proteínas/metabolismo , Proteínas/farmacología , Piroptosis/efectos de los fármacos , Piroptosis/inmunologíaRESUMEN
Thirteen benzopyran derivatives were synthesized and their activity stimulating the differentiation of preadipocytes into adipocytes were evaluated with 3T3-L1 cells. Compound 8 was also tested for its hypoglycemic activity on db diabetes mice model. Results indicated that compounds 3, 8 and 11 exhibited strong differentiation-stimulating activity on 3T3-L1 cells model, and compound 8 can reduce the blood-sugar level of db diabetes mice dramatically.
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Animales , Ratones , Células 3T3-L1 , Adipocitos , Benzopiranos , Química , Farmacología , Glucemia , Metabolismo , Diferenciación Celular , Diabetes Mellitus Experimental , Sangre , Hipoglucemiantes , Química , FarmacologíaRESUMEN
Ten novel compounds were designed and synthesized on the basis of compound 1, their insulin-sensitizing activities were evaluated in 3T3-L1 cells. Results showed that compound 10 exhibited strong differentiation-stimulating activity on 3T3-L1 cells model, which indicated that compound 10 may possess well insulin-sensitizing activity.
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Animales , Ratones , Células 3T3-L1 , Benzopiranos , Farmacología , Diseño de Fármacos , Hipoglucemiantes , Farmacología , Insulina , FarmacologíaRESUMEN
To design and synthesis a series of novel L-amino acid esters prodrugs of acyclic nucleoside phosphonates with more potent anti-HBV activity, adefovir dipivoxil was used as lead compound, according to the results of enhanced oral bioavailability and antiviral activities of nucleoside L-amino acid ester prodrugs. Eleven novel L-amino acid ester prodrugs of acyclic nucleoside phosphonates were designed and synthesized, their anti-HBV activities were evaluated in HepG2 2.2.15 cells. Eight compounds exhibited antiviral activity, and compound 11 showed the most potent anti-HBV activity and highest selective index in vitro (EC50 0.0952 micromol x L(-1), SI 69523). Moreover, by analyzing the primary structure and activity relationship of these compounds, it could be suggested that L-amino acid ester strategy has significant potential in the acyclic nucleoside phosphonates prodrug design.
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Humanos , Aminoácidos , Química , Antivirales , Farmacología , Línea Celular Tumoral , Virus de la Hepatitis B , Neoplasias Hepáticas , Patología , Virología , Nucleósidos , Farmacología , Organofosfonatos , Farmacología , Profármacos , FarmacologíaRESUMEN
The specific expression of TGH and HSL genes in different tissues of Bamei pig was investigated by RT-PCR and Western blot in this study. The result of RT-PCR showed that the expression of HSL could be detected in all these seven tissues examined, and which was higher expressed in fat, lower in heart, liver, lung, spleen and kidney. Expression of TGH gene could also be detected in seven tissues, and higher in liver and fat, lower in heart and kidney and lowest in spleen and lung. The result of Western blot showed that, HSL gene was highest expressed in epiploica fat and subcutaneous fat, higher in other tissues, but couldn' t be detected in kidney. Expression of TGH was detected in epiploica fat, subcutaneous fat, liver, lung and spleen, and highest in fat and liver, but it hadn't be found in heart and kidney. These results suggested that both HSL and TGH could be regulated by post-transcriptional, and their function was involved in different tissues.
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Animales , Masculino , Regulación de la Expresión Génica , Lipasa , Genética , Metabolismo , Esterol Esterasa , Genética , Metabolismo , Porcinos , Distribución TisularRESUMEN
<p><b>OBJECTIVE</b>To study the effect of emodin (EMO) on the proliferation and differentiation of rat preadipocytes.</p><p><b>METHOD</b>Separating and culturing rat preadipocytes, grouping the wells that preadipocytes were growing according to certain concentration such as 0, 5, 10, 20, 40, 80, 160 micromol x L(-1) randomly, MTT spectrophotometry and flow cytometry (FCM) were adopted to determine the effect of EMO on proliferation of rat preadipocytes. The accumulation of TG (triglyceride) in adipocytes was assayed by oil red O staining, and the morphological changes of the adipocytes were determined by morphology observation.</p><p><b>RESULT</b>EMO in the range of 20-160 micromol x L(-1) could inhibit the proliferation and differentiation of preadipocytes in a dose and time dependent manner, and induce apoptosis of preadipocytes in a certain degree.</p><p><b>CONCLUSION</b>EMO should have a potential to serve as a fat-reducing drug.</p>
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Animales , Masculino , Ratas , Adipocitos , Biología Celular , Apoptosis , Ciclo Celular , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Relación Dosis-Respuesta a Droga , Emodina , Farmacología , Distribución Aleatoria , Ratas Sprague-DawleyRESUMEN
Total RNA was isolated from kidney of BaMei pig, a local strain of Chinese pig, and then the cDNA sequence of SOCS-2 gene was cloned by RT-PCR (GenBank accepted number is EF121242). Then the cloned SOCS-2 gene was inserted into PMD19-T vector by T/A cloning, transformed into DH-5alpha, tested by PCR and sequenced. The data show that the homology of the cloned porcine SOCS-2, including 822 bp, is more than 93% and that of the deduced amino acid sequence is 89% when compared with human, rat and mice. And the molecular weight of SOCS-2 protein is about 22.25 kD and PI is 8.03. The cloning of SOCS-2 gene is useful for the further research on the molecular mechanism by which regulating growth and development of organism.