ABSTRACT
Objective:To study the regulatory effect of Wumen-Yiji powder on 5-hydroxytryptamine (5-HT) signal transduction system in intestine and hypothalamus of diarrhea irritable bowel syndrome (IBS-D) model rats. Methods:Sixty male SD rats were randomly divided into blank group (10 rats) and diarrhea irritable bowel syndrome group (50 rats). The diarrhea irritable bowel syndrome group formed the diarrhea irritable bowel syndrome model after 2 weeks of senna leaf gavage and restraint stress. They were randomly divided into model group, deshute group (1.5 mg/kg), low, medium and high dose group of Wumen-Yiji San (6, 12, 24 g/kg), with 10 rats in each group. After continuous administration for 2 weeks, the contents of 5-HT in serum, colon and hypothalamus were detected by ELISA; HE staining was used to observe the pathological changes of colon in each group. The protein and mRNA levels of tryptophan hydroxylase 1 (TPH-1), serotonin receptor 3 (5-HT3R), serotonin receptor 4 (5-HT4R), serotonin transporter (SERT) in colon and hypothalamus were detected by Western blot and RT-PCR, respectively. Results:Compared with the model group, the pathological morphology of colon in each treatment group was improved. Compared with the model group, the level of 5-HT in serum and colon significantly decreased ( P<0.05), and the level of 5-HT in hypothalamus of rats in the low, medium, high dose group of Wumen-Yiji San significantly increased ( P<0.05). The expression of TPH-1, 5-HT3R and 5-HT4R protein significantly decreased ( P<0.05), and the expression of SERT protein in the medium, high dose group of Wumen-Yiji San significantly increased ( P<0.05). The expression of TPH-1, 5-HT3R and 5-HT4R protein in hypothalamus increased ( P<0.05), and the expression of SERT protein in the high dose group of Wumen-Yiji San significantly decreased ( P<0.05). The mRNA levels of TPH-1 (4.778 ± 0.604, 3.278 ± 0.668, 1.670 ± 0.361 vs. 6.877 ± 0.148), 5-HT3R (3.807 ± 0.463, 2.697 ± 0.455, 1.132 ± 0.136 vs. 6.322 ± 0.778), 5-HT4R (4.521 ± 0.234, 2.801 ± 0.351, 1.331 ± 0.142 vs. 6.741 ± 0.293) in colon tissue of low, medium and high dose groups of Wumen-Yiji San decreased ( P<0.05). The level of 5-HT4R mRNA (0.616 ± 0.208, 0.726 ± 0.226 vs. 0.521 ± 0.062) increased ( P<0.05), and the level of SERT mRNA (1.563 ± 1.023 vs. 2.612 ± 1.035) in medium, high dose group of Wumen-Yiji San decreased ( P<0.05). Conclusion:The result showed that Wumen-Yiji San could regulate the expression of 5-HT signaling system relating proteins and mRNA in the colon and hypothalamus of IBS-D rats within a certain dose range, so as to improve the symptoms of IBS-D.
ABSTRACT
Staphylococcus utilizes vancomycin-resistance associated sensor/regulator ( VraSR) , a two-component signal transduction system ( TCS) , to sense and respond to cell wall damage and to adapt to environmental changes through regulating transcriptions of downstream genes. It has been indicated that VraSR can regulate the transcription of a series of genes involved in the synthesis of peptidoglycan, drug re-sistance, and virulence in Staphylococcus aureus ( S. aureus) . A similar two-component system, VraSR, is also present in Staphylococcus epidermidis ( S. epidermidis ) , sharing a high homology with the VraSR of S. aureus. Little is known about the functions of VraSR in S. epidermidis and it is not yet clear what the simi-larities and differences in resistance and pathogenicity are. Based on the previous work of our group, a brief review on the regulation mechanism of staphylococcal VraSR was performed.
ABSTRACT
Vibrio vulnificus is an estuarine bacterium that opportunistically infects humans with underlying hepatic diseases, heavy alcohol drinking habits, and other immunocompromised conditions. A locus in the V. vulnificus genome was cloned and sequenced, which showed similarities to the bacterial two-component signal transduction system. The locus encoded a putative sensor kinase, designated vvgS, and a divergently transcribed putative response regulator, designated vvgR. VvgS was predicted to be a protein belonging to the tripartite hybrid sensor kinase subfamily such as ArcB and BvgS. VvgR showed similarities to well known response regulators. An insertional mutation of vvgS in a V. vulnificus strain led to a remarkable decrease of cytotoxicity to HeLa cells, while the production of exotoxins, the hemolysin and the protease, slightly increased by the vvgS mutation. Adhesion to HeLa cells only slightly decreased. However, the vvgS mutation had no effect on the motility of V. vulnificus. The vvgS mutation resulted in a significant decrease of lethality to mice. These results indicate that vvgRS two-component system plays a very important role in regulating novel virulence factor(s) of V. vulnificus associated with cytotoxicity other than hemolysin and protease during the infection process.
Subject(s)
Animals , Humans , Mice , Alcohol Drinking , Clone Cells , Exotoxins , Genome , HeLa Cells , Phosphotransferases , Signal Transduction , Vibrio vulnificus , Vibrio , VirulenceABSTRACT
Vibrio vulnificus is an estuarine bacterium that opportunistically infects humans with underlying hepatic diseases, heavy alcohol drinking habits, and other immunocompromised conditions. A locus in the V. vulnificus genome was cloned and sequenced, which showed similarities to the bacterial two-component signal transduction system. The locus encoded a putative sensor kinase, designated vvgS, and a divergently transcribed putative response regulator, designated vvgR. VvgS was predicted to be a protein belonging to the tripartite hybrid sensor kinase subfamily such as ArcB and BvgS. VvgR showed similarities to well known response regulators. An insertional mutation of vvgS in a V. vulnificus strain led to a remarkable decrease of cytotoxicity to HeLa cells, while the production of exotoxins, the hemolysin and the protease, slightly increased by the vvgS mutation. Adhesion to HeLa cells only slightly decreased. However, the vvgS mutation had no effect on the motility of V. vulnificus. The vvgS mutation resulted in a significant decrease of lethality to mice. These results indicate that vvgRS two-component system plays a very important role in regulating novel virulence factor(s) of V. vulnificus associated with cytotoxicity other than hemolysin and protease during the infection process.
Subject(s)
Animals , Humans , Mice , Alcohol Drinking , Clone Cells , Exotoxins , Genome , HeLa Cells , Phosphotransferases , Signal Transduction , Vibrio vulnificus , Vibrio , VirulenceABSTRACT
0.05).After treatment,the nipple height was increased in the model group(P
ABSTRACT
The signal transduction system induced by epidermal growth factor receptor(EGFR) regulates cell cycle ,modulates cell growth and differentiation and improves damage repair.The overexpression of EGFR in several epithelial tumors including non small cell lung cancer(NSCLC) forecasts low survival rate, poor prognosis and metastasis. Thereby EGFR can be a potential target for gene therapy. Tyrosine kinase inhibitors(TKIs) selectively inhibit tyrosine kinase activity,supress tumor growth ,and increase the sensitivity of radio chemotherapy. [
ABSTRACT
Oncogenes are known to be involved in normal cellular growth and proliferation as well as in carcinogenesis. It is reported that stimulation of quiescent cells with growth factors makes the oncogenes produce protein products as an early and immediate response, and these protein products induce or control the cell growth. Among those oncogenes, c-fos and c-myc are well known for its generalized expressions. An experiment was performed in order to prove the hypothesis that oncogene expressions would have the same pattern with that of cellular growth by growth factors in cultured normal rat thyroid cell line(FRTL-5). Ribonucleic acids of FRTL-5 cells were extracted time-sequentially at 15, 30, 60 and 120 minutes after administration of growth factors to the media of quiescent FRTL-5 cells. Extracted ribonucleic acids were blotted to the nitrocellulose membrane, and then hybridized with radiolabelled c-fos and c-myc oligonucleotide probes, and -actin probes. Hybridized dot blots on nitrocellulose membrane were autoradiographed on X-ray films, and the amount of radioactivity were measured by densitometry. Densitometric readings were used as the indices of oncogene expressions. The concentrations of TSH, IGF-I and IgG from patients with Graves' disease, which showed the maximum expressions of c-fos and c-myc in quiescent FRTL-5 cells, were TSH 10 mU/ml, IGF-I 100 ng/ml and IgG from patients with Graves' disease 1 mg/ml, respectively. Expressions of c-fos and c-myc were more prominent in combined administrations of TSH and IGF-I, or IgG from patients with Graves' disease and IGF-I than in case of TSH and IgG from patients with Graves' disease. IgG from patients with primary myxedema suppressed oncogene expressions provoked by TSH or IgG from patients with Graves' disease, but not by IGF-I. Expressions of c-fos and c-myc were more prominent by the combined administration of TSH with TPA which stimulated the phosphoinositide turnover-protein kinase C-calcium system than by those of TSH with dBcAMP, forskolin, IBMX or cholera toxin which stimulated adenylate cyclase system. From the above results, following conclusions were obtained. 1) Expressions of c-fos and c-myc by growth factors have similar patterns with those of cell growth by growth factors in FRTL-5 cells. 2) It is suggested that the actions of TSH and IgG from patients with Graves' disease would be manifested through the same signal transduction system, and that of IGF-I would be manifested through its own, but the oncogenes would be expressed mainly through adenylate cyclase system.
Subject(s)
Animals , Humans , Rats , 1-Methyl-3-isobutylxanthine , Adenylyl Cyclases , Bucladesine , Carcinogenesis , Cell Line , Cholera Toxin , Colforsin , Collodion , Densitometry , Genes, myc , Graves Disease , Immunoglobulin G , Insulin-Like Growth Factor I , Intercellular Signaling Peptides and Proteins , Membranes , Myxedema , Oligonucleotide Probes , Oncogenes , Phosphotransferases , Radioactivity , Reading , RNA , Signal Transduction , Thyroid Gland , X-Ray FilmABSTRACT
This study was performed to prove the hypothesis that oncogene expressions would have the same patterns with those of cellular growth to growth factors in FRTL-5 cells. Ribonucleic acids of FRTL-5 were extracted at 15', 30', 60' and 120' after administration of growth factors to quiescent FRTL-5, and blotted to the nitrocellulose membrane. They were hybridized with radiolabelled c-fos, c-myc and beta-actin probes. Hybridized dot blots were autoradiographed and the amount of radioactivity was measured by densitometry. Densitometric readings were used as the indices of oncogene expressions. Expressions of c-fos and c-myc were more prominent in combined administrations of TSH (10 mU/ml) and IGF-I (100 ng/ml) or IgG of Graves' disease (Graves' IgG; 1 mg/ml) and IGF-I than in combined administration of TSH and Graves' IgG. IgG of primary myxedema suppressed oncogene expressions by TSH or Graves' IgG, but not by IGF-I. From the above results, it was suggested that expressions of c-fos and c-myc to growth factors would have similar patterns with those of cell growth to growth factors in FRTL-5, and the actions of TSH and Graves' IgG would be manifested through same signal transduction system, but IGF-I would be manifested by its own.
Subject(s)
Rats , Animals , Cell Division/drug effects , Cell Line/cytology , Gene Expression/drug effects , Graves Disease/immunology , Growth Substances/genetics , Immunoglobulin G/pharmacology , Insulin-Like Growth Factor I/pharmacology , Myxedema/immunology , Proto-Oncogene Proteins c-fos/genetics , Proto-Oncogene Proteins c-myc/genetics , RNA/analysis , Rats, Inbred F344 , Thyroid Gland/cytology , Thyrotropin/pharmacology , Time FactorsABSTRACT
To construct gene knock-out mutant of a two-component signal transduction system named 2148hk/rr in Streptococcus suis type 2 virulent strain 05ZYH33.Recombinant gene knock-out vector was constructed consisting of Spc~r cassette with flanking homology regions to the target genes,the isogenic 2148hk/rr-deficient mutant was screened by allelic replacement.PCR analysis and Southern hybridization confirmed that the coding genes of 2148hk/rr were replaced completely by spc~r cassette.Conclusion The mutant of 05ZYH33 2148hk/rr system was successfully constructed,which laid the foundation for farther research on the role of this two-component signal transduction system during infection.