To determine whether the cyclic adenosine monophosphate (cAMP) mediated protein kinase signal transduction pathway is involved in the pyrogenic action of corticotropin releasing hormone (CRH) in rats.Methods Corticot...To determine whether the cyclic adenosine monophosphate (cAMP) mediated protein kinase signal transduction pathway is involved in the pyrogenic action of corticotropin releasing hormone (CRH) in rats.Methods Corticotropin releasing hormone, 2', 3 '-dideoxyadenosine (DDA) and adenosine-3', 5'(cyclic) monophosphorothionate, Rp-lsomer (Rp-cAMPS), were administered intracerebroventricularly (i.c.v.). The colonic temperature was measured using a thermistor, and the content of cAMP in the hypothalamus was determined by radioimmunoassay. Hypethalemic incubation was used to assess the effects of CRH on the content of cAMP in the hypothalamus in vitro.Results Microinjection (i.c.v.) of CRH (2.5 μg, 5.0 μg and 10 μg) caused increases in colonic temperature and the hypothalemus cAMP level in conscious rats. CRH increased hypothalemus cAMP level in vitro. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (i. c. v. ) of an adenylate cyclase inhibitor, DDA (30 μg), or an inhibitor of cAMP-dependent protein kinase, Rp-cAMPS (15 μg).Conclusion cAMP mediates the pyrogenic action of centrally administered of CRH in rats, and protein kinase A may play an important role in the central CRH-induced fever. The cAMP-dependent protein kinase signal transduction pathway may be involved in the central mechanisms of the pyrogenic action of CRH in rats.展开更多
Objective To examine whether urocortin is produced locally to regulate utero-placental vascular tone during pregnancy.Methods We examined the distribution of urocortin in human placenta, fetal membranes and uterine ...Objective To examine whether urocortin is produced locally to regulate utero-placental vascular tone during pregnancy.Methods We examined the distribution of urocortin in human placenta, fetal membranes and uterine tissue at term in the presence and absence of labor using a urocortin antibody produced in our laboratory and the immunoperoxidase staining method. Subsequently, we tested urocortin secretion from chorio-decidual cells in vitro using an immunoblot technique. Then, we tested whether urocortin is present in maternal plasma throughout gestation using a radioimmunoassay. A Sephadex G-50 column was used to examine whether immunoreactive urocortin (IR-urocortin) in maternal plasma is the same as synthetic urocortin.Results IR-urocortin was observed in vascular smooth muscle of myometrium decidual stromal cells, syncytiotrophoblast and amnion epithelium. No differences in staining intensity for urocortin were detected between tissues obtained in the absence or presence of labor. Staining intensity for IR-urocortin was greatest in the decidua, suggesting this may be the main site of urocortin production. Positive staining for urocortin was observed in 40% of chorio-decidual cells with 34% of these cells secreting urocortin under basal conditions. Urocortin was detectable in maternal plasma from 16 weeks gestation and concentrations did not change as gestation progressed. IR-urocortin in the maternal plasma eluted from a Sephadex G-50 column at the same site as synthetic urocortin and had a calculated retention co-efficient of 0.44.Conclusion This study indicates that urocortin is produced by the decidua during human pregnancy and is detectable in maternal plasma. These data are consistent with the hypothesis that urocortin is produced locally by the decidua and may act to regulate utero-placental blood flow.展开更多
In trauma, infection and hemorrhagic shock derived stress, primary and secondary injury may result in severe derangement in the internal environment. The abnormal changes of immune-mediated inflammation interfere its ...In trauma, infection and hemorrhagic shock derived stress, primary and secondary injury may result in severe derangement in the internal environment. The abnormal changes of immune-mediated inflammation interfere its pathogenesis and development directly. In recent years, various aspects of neuroendocrine responses, especially the regulatory effects of hypothalamic-pituitary-adrenal and sympathetico-adrenomedullary axes in inflammatory diseases have been the focus of research. Most importantly,corticotropin-releasing hormone （CRH） acts as a key player in the regulation of interactions between neuroendocrine and immunity both directly and indirectly. The paper summarized the recent development of CRH in the immune-mediated inflammation.展开更多
The bed nucleus of the stria terminalis(BNST) plays a critical role in regulating anxiety, yet the involved specific cell types and their connections functioning in anxiety-related behaviors remains elusive. Here we i...The bed nucleus of the stria terminalis(BNST) plays a critical role in regulating anxiety, yet the involved specific cell types and their connections functioning in anxiety-related behaviors remains elusive. Here we identified two cell subpopulations—corticotropin-releasing hormone-positive(CRH+) and protein kinase C-δ-positive(PKC-δ+) neurons—each displayed discrete emotionally valenced behaviors in the anterior-dorsal BNST(ad BNST). Using whole-cell patch-clamp recordings and virus-assisted circuit tracing techniques, we delineated the local and long-range connectivity networks in a cell-type-specific manner. The results show that the CRH+ and PKC-δ+ neurons received inputs from similar brain regions and exhibited significant differences in the downstream projection density. In addition, in vivo calcium imaging as well as gain-and loss-of-function studies characterized the physiological response properties and the functional heterogeneities in modulating anxiety, further suggesting the similarity and individuality between the two ad BNST cell types. These results provide novel insights into the circuit architecture of ad BNST neurons underlying the functionally specific neural pathways that relate to anxiety disorders.展开更多
OBJECTIVE: We wished to study the impact of Chaihushugan San(CSS) on the behavior of perimenopausal rats with liver-Qi stagnation(LQS) and to investigate the effect of CSS on signal transduction of the Raf/mitogen-act...OBJECTIVE: We wished to study the impact of Chaihushugan San(CSS) on the behavior of perimenopausal rats with liver-Qi stagnation(LQS) and to investigate the effect of CSS on signal transduction of the Raf/mitogen-activated protein kinase(MEK)/extracellular signal-regulated kinase(ERK) cascade in the hippocampi of rats induced by immobilization.METHODS: Twenty 52-week-old female rats were divided into two groups by the random number table method: model control group(MCG) and CSS group(CSSG), with 10 rats in each group.Ten-week-old female rats were used as the normal control group(NCG). CSS effects were assessed using rats exposed to immobilization stress by measuring body weight and sucrose consumption, serum hormone levels, and observing performance in the open field test(OFT). Molecular mechanisms were examined by measuring the effect of CSS on expression of Raf1, MEK1/2 and ERK1/2 m RNA in hippocampi using quantitative real-time polymerase chain reaction and by measuring levels of these proteins and related phospho-proteins using Western blotting.RESULTS: Perimenopausal rats with LQS had decreased locomotor activity; reduced sucrose consumption; and increased serum levels of corticotropin releasing hormone(CRH) and corticosterone(CORT). Activation of hippocampal Raf/MEK/ERK cascade was suppressed significantly in the MCG,and activation was increased after 21 days of CSS treatment.CONCLUSION: CSS has significant effects upon relief of the symptoms of LQS in immobilization-induced rats. The mechanism underlying this action might(at least in part) be mediated by reversal of disruption of the Raf/MEK/ERK pathway.展开更多
Objective： To explore the changes of Treg-Thl 7 balance influenced by corticosterone, major ef- fect hormone ofhypothalamic-pituitary-adrenal （HPA） axis under running stress. Methods： Atotal of 25 corticotropin-r...Objective： To explore the changes of Treg-Thl 7 balance influenced by corticosterone, major ef- fect hormone ofhypothalamic-pituitary-adrenal （HPA） axis under running stress. Methods： Atotal of 25 corticotropin-releasing hor- mone （CRH） wildtype （CRH＋/＋） and knockout （CRH-/-） mice were adopt and divided into 4 groups as follows： CRH＋/＋ ctrl, CRH＋/＋ stress, CRH-/- ctrl and CRH-/- stress. All mice in stress groups were under 2 h running. After 1 h, blood plasma in all groups was collected and the ex- pression of corticosterone and IL- 17A was detected by ELISA. Meanwhile, unicell suspensions of peripheral lymph node and spleen in each group were prepared too and stained by PE-CD4 and FITC-CD25, then the changes of Treg （CD4＋CD25＋） in different groups were checked by flow cytometry; all data were statistically analyzed by the software of WinMDI 2.9, SPSS 11.5, Origin 7.5 and Matlab 2-D and 3-D plot function. Results： The levels of corticosterone were signifi- cantly higher in stress groups than that in correspond- ing control groups （P〈0.05）, especially in CRH＋/＋ stress group （P〈0.01）. However, the changes of Tregs were not obvious between stress groups and control groups with respective genotypes （P〈0.05）. Compared with that in CRH＋/＋ control group, the ratio of Treg and the expres- sion of IL-17A in CRH-/- stress group were significantly higher than those in control group （P〈0.05）. Combined with the expression levels of corticosterone, Treg and Thl7, our study suggests that endogenous glucocorti- cold with basal level may cause the changes in Treg- Thl7 balance. Moreover, as the corticosterone level increases, the expression of Treg and Thl7 appears to manifest antagonistic fluctuant status with a rising ten- dency in general. Conclusion： Endogenous glucocorticoid under early stage of stress may increase the function of T lymphocyte immunity to some extent.展开更多
Objective： To observe the effect of moxibustion on the protein and mRNA expressions of corticotropin-releasing factor （CRF） and corticotropin-releasing factor receptor 2 （CRFR1） in hypothalamus of trinitrobenzene...Objective： To observe the effect of moxibustion on the protein and mRNA expressions of corticotropin-releasing factor （CRF） and corticotropin-releasing factor receptor 2 （CRFR1） in hypothalamus of trinitrobenzene sulfonic acid （TNBS）-induced experimental colitis rats, and to explore the central mechanisms of moxibustion in improving visceral pain and the pain-related emotions in experimental colitis rats. Methods： Thirty-six Sprague-Dawley （SD） rats were randomly divided into a normal group （NG）, a model group （MG）, a herb-partitioned moxibustion group （HPMG） and a sham herb-partitioned moxibustion group （SHPMG）. Except the NG, rats in the remaining three groups all received TNBS enema to establish experimental colitis models. The HPMG received herb-partitioned moxibustion （HPM） at bilateral Tianshu （ST 25） and Qihai （CV 6） for intervention; for the SHPMG, the herbal cakes and moxa cones were only placed on the acupoints but not ignited; rats in the MG and NG were only fixed in the same way as those in the HPMG but did not receive any treatment. At the end of the intervention, the abdominal withdrawal reflex （AWR） score, the open field test （OFT） score and the elevated plus maze （EPM） score were observed to measure the changes in visceral pain and pain-related emotions of the rats. The enzyme-linked immunosorbent assay （ELISA） was used to examine the expressions of CRF and CRFR1 proteins in hypothalamus; the fluorescence-based quantitative polymerase chain reaction （PCR） was used to detect the expressions of CRF and CRFR1 mRNAs in hypothalamus. Results： Compared with the NG, the AWR score increased significantly and the OFT and EPM scores dropped significantly in the MG （all P〈0.05）, and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs increased significantly （all P〈0.01）. Compared with the MG and SHPMG, the AWR score dropped significantly and the OFT and EPM scores increased significantly in the HPMG （all P〈0.01）, and the expressions of hypothalamic CRF and CRFRI proteins and mRNAs decreased significantly （all P〈0.05）. There were no significant differences between the MG and the SHPMG （all P〉0.05）. Conclusion： HPM can down-regulate the abnormally increased expressions of CRF and CRFR1 proteins and mRNAs in hypothalamus of the TNBS-induced experimental colitis rats, which is plausibly one of its action mechanisms in mitigating visceral pain and the pain-related emotions in the experimental colitis rats.展开更多
基金ThisworkwassupportedbytheNationalNaturalScienceFoundationofChina (No 3970 0 0 5 5 )
文摘To determine whether the cyclic adenosine monophosphate (cAMP) mediated protein kinase signal transduction pathway is involved in the pyrogenic action of corticotropin releasing hormone (CRH) in rats.Methods Corticotropin releasing hormone, 2', 3 '-dideoxyadenosine (DDA) and adenosine-3', 5'(cyclic) monophosphorothionate, Rp-lsomer (Rp-cAMPS), were administered intracerebroventricularly (i.c.v.). The colonic temperature was measured using a thermistor, and the content of cAMP in the hypothalamus was determined by radioimmunoassay. Hypethalemic incubation was used to assess the effects of CRH on the content of cAMP in the hypothalamus in vitro.Results Microinjection (i.c.v.) of CRH (2.5 μg, 5.0 μg and 10 μg) caused increases in colonic temperature and the hypothalemus cAMP level in conscious rats. CRH increased hypothalemus cAMP level in vitro. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (i. c. v. ) of an adenylate cyclase inhibitor, DDA (30 μg), or an inhibitor of cAMP-dependent protein kinase, Rp-cAMPS (15 μg).Conclusion cAMP mediates the pyrogenic action of centrally administered of CRH in rats, and protein kinase A may play an important role in the central CRH-induced fever. The cAMP-dependent protein kinase signal transduction pathway may be involved in the central mechanisms of the pyrogenic action of CRH in rats.
文摘Objective To examine whether urocortin is produced locally to regulate utero-placental vascular tone during pregnancy.Methods We examined the distribution of urocortin in human placenta, fetal membranes and uterine tissue at term in the presence and absence of labor using a urocortin antibody produced in our laboratory and the immunoperoxidase staining method. Subsequently, we tested urocortin secretion from chorio-decidual cells in vitro using an immunoblot technique. Then, we tested whether urocortin is present in maternal plasma throughout gestation using a radioimmunoassay. A Sephadex G-50 column was used to examine whether immunoreactive urocortin (IR-urocortin) in maternal plasma is the same as synthetic urocortin.Results IR-urocortin was observed in vascular smooth muscle of myometrium decidual stromal cells, syncytiotrophoblast and amnion epithelium. No differences in staining intensity for urocortin were detected between tissues obtained in the absence or presence of labor. Staining intensity for IR-urocortin was greatest in the decidua, suggesting this may be the main site of urocortin production. Positive staining for urocortin was observed in 40% of chorio-decidual cells with 34% of these cells secreting urocortin under basal conditions. Urocortin was detectable in maternal plasma from 16 weeks gestation and concentrations did not change as gestation progressed. IR-urocortin in the maternal plasma eluted from a Sephadex G-50 column at the same site as synthetic urocortin and had a calculated retention co-efficient of 0.44.Conclusion This study indicates that urocortin is produced by the decidua during human pregnancy and is detectable in maternal plasma. These data are consistent with the hypothesis that urocortin is produced locally by the decidua and may act to regulate utero-placental blood flow.
基金This work was supported by the Special Funds for Major State Basic Research Projects （2005CB522600）, the Eleventh Five-Year Science and Technology Project of PLA （06G080）, and Natural Science Foundation Project of CQCSTC （2009BB5316）
文摘In trauma, infection and hemorrhagic shock derived stress, primary and secondary injury may result in severe derangement in the internal environment. The abnormal changes of immune-mediated inflammation interfere its pathogenesis and development directly. In recent years, various aspects of neuroendocrine responses, especially the regulatory effects of hypothalamic-pituitary-adrenal and sympathetico-adrenomedullary axes in inflammatory diseases have been the focus of research. Most importantly,corticotropin-releasing hormone （CRH） acts as a key player in the regulation of interactions between neuroendocrine and immunity both directly and indirectly. The paper summarized the recent development of CRH in the immune-mediated inflammation.
基金This work was supported by the National Natural Science Foundation of China(31671105)Science Fund for Creative Research Group of China(61721092),“National Basic Research Program of China”(973 program 2015CB755603)Director Fund of the Wuhan National Laboratory for Optoelectronics and partly supported by the open funds of the State Key Laboratory of Medical Neurobiology,Fudan University.
文摘The bed nucleus of the stria terminalis(BNST) plays a critical role in regulating anxiety, yet the involved specific cell types and their connections functioning in anxiety-related behaviors remains elusive. Here we identified two cell subpopulations—corticotropin-releasing hormone-positive(CRH+) and protein kinase C-δ-positive(PKC-δ+) neurons—each displayed discrete emotionally valenced behaviors in the anterior-dorsal BNST(ad BNST). Using whole-cell patch-clamp recordings and virus-assisted circuit tracing techniques, we delineated the local and long-range connectivity networks in a cell-type-specific manner. The results show that the CRH+ and PKC-δ+ neurons received inputs from similar brain regions and exhibited significant differences in the downstream projection density. In addition, in vivo calcium imaging as well as gain-and loss-of-function studies characterized the physiological response properties and the functional heterogeneities in modulating anxiety, further suggesting the similarity and individuality between the two ad BNST cell types. These results provide novel insights into the circuit architecture of ad BNST neurons underlying the functionally specific neural pathways that relate to anxiety disorders.
基金Supported by the National Natural Sciences Foundations of China(Study on the Molecular Mechanism of Formation of Liver-Qi Stagnation in Perimenopausal Syndrome Based on the Neuroendocrine Axis and Ras/Raf/MEK/ERK Pathway,No.81302889Study on the Mechanism of Liver-Qi Stagnation in Perimenopausal Syndrome Based on "Women Easily Qi-Stagnated" and Ca2+/Camp Pathway Mediated by the Estrogen Receptor,No.81173203)
文摘OBJECTIVE: We wished to study the impact of Chaihushugan San(CSS) on the behavior of perimenopausal rats with liver-Qi stagnation(LQS) and to investigate the effect of CSS on signal transduction of the Raf/mitogen-activated protein kinase(MEK)/extracellular signal-regulated kinase(ERK) cascade in the hippocampi of rats induced by immobilization.METHODS: Twenty 52-week-old female rats were divided into two groups by the random number table method: model control group(MCG) and CSS group(CSSG), with 10 rats in each group.Ten-week-old female rats were used as the normal control group(NCG). CSS effects were assessed using rats exposed to immobilization stress by measuring body weight and sucrose consumption, serum hormone levels, and observing performance in the open field test(OFT). Molecular mechanisms were examined by measuring the effect of CSS on expression of Raf1, MEK1/2 and ERK1/2 m RNA in hippocampi using quantitative real-time polymerase chain reaction and by measuring levels of these proteins and related phospho-proteins using Western blotting.RESULTS: Perimenopausal rats with LQS had decreased locomotor activity; reduced sucrose consumption; and increased serum levels of corticotropin releasing hormone(CRH) and corticosterone(CORT). Activation of hippocampal Raf/MEK/ERK cascade was suppressed significantly in the MCG,and activation was increased after 21 days of CSS treatment.CONCLUSION: CSS has significant effects upon relief of the symptoms of LQS in immobilization-induced rats. The mechanism underlying this action might(at least in part) be mediated by reversal of disruption of the Raf/MEK/ERK pathway.
基金National Natural Science Foundation of China，Natural Science Foundation Project of CQ CSTC，Foundation for Returned Scholars Sponsored by Department of Human Resources and Social Security of China
文摘Objective： To explore the changes of Treg-Thl 7 balance influenced by corticosterone, major ef- fect hormone ofhypothalamic-pituitary-adrenal （HPA） axis under running stress. Methods： Atotal of 25 corticotropin-releasing hor- mone （CRH） wildtype （CRH＋/＋） and knockout （CRH-/-） mice were adopt and divided into 4 groups as follows： CRH＋/＋ ctrl, CRH＋/＋ stress, CRH-/- ctrl and CRH-/- stress. All mice in stress groups were under 2 h running. After 1 h, blood plasma in all groups was collected and the ex- pression of corticosterone and IL- 17A was detected by ELISA. Meanwhile, unicell suspensions of peripheral lymph node and spleen in each group were prepared too and stained by PE-CD4 and FITC-CD25, then the changes of Treg （CD4＋CD25＋） in different groups were checked by flow cytometry; all data were statistically analyzed by the software of WinMDI 2.9, SPSS 11.5, Origin 7.5 and Matlab 2-D and 3-D plot function. Results： The levels of corticosterone were signifi- cantly higher in stress groups than that in correspond- ing control groups （P〈0.05）, especially in CRH＋/＋ stress group （P〈0.01）. However, the changes of Tregs were not obvious between stress groups and control groups with respective genotypes （P〈0.05）. Compared with that in CRH＋/＋ control group, the ratio of Treg and the expres- sion of IL-17A in CRH-/- stress group were significantly higher than those in control group （P〈0.05）. Combined with the expression levels of corticosterone, Treg and Thl7, our study suggests that endogenous glucocorti- cold with basal level may cause the changes in Treg- Thl7 balance. Moreover, as the corticosterone level increases, the expression of Treg and Thl7 appears to manifest antagonistic fluctuant status with a rising ten- dency in general. Conclusion： Endogenous glucocorticoid under early stage of stress may increase the function of T lymphocyte immunity to some extent.
文摘Objective： To observe the effect of moxibustion on the protein and mRNA expressions of corticotropin-releasing factor （CRF） and corticotropin-releasing factor receptor 2 （CRFR1） in hypothalamus of trinitrobenzene sulfonic acid （TNBS）-induced experimental colitis rats, and to explore the central mechanisms of moxibustion in improving visceral pain and the pain-related emotions in experimental colitis rats. Methods： Thirty-six Sprague-Dawley （SD） rats were randomly divided into a normal group （NG）, a model group （MG）, a herb-partitioned moxibustion group （HPMG） and a sham herb-partitioned moxibustion group （SHPMG）. Except the NG, rats in the remaining three groups all received TNBS enema to establish experimental colitis models. The HPMG received herb-partitioned moxibustion （HPM） at bilateral Tianshu （ST 25） and Qihai （CV 6） for intervention; for the SHPMG, the herbal cakes and moxa cones were only placed on the acupoints but not ignited; rats in the MG and NG were only fixed in the same way as those in the HPMG but did not receive any treatment. At the end of the intervention, the abdominal withdrawal reflex （AWR） score, the open field test （OFT） score and the elevated plus maze （EPM） score were observed to measure the changes in visceral pain and pain-related emotions of the rats. The enzyme-linked immunosorbent assay （ELISA） was used to examine the expressions of CRF and CRFR1 proteins in hypothalamus; the fluorescence-based quantitative polymerase chain reaction （PCR） was used to detect the expressions of CRF and CRFR1 mRNAs in hypothalamus. Results： Compared with the NG, the AWR score increased significantly and the OFT and EPM scores dropped significantly in the MG （all P〈0.05）, and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs increased significantly （all P〈0.01）. Compared with the MG and SHPMG, the AWR score dropped significantly and the OFT and EPM scores increased significantly in the HPMG （all P〈0.01）, and the expressions of hypothalamic CRF and CRFRI proteins and mRNAs decreased significantly （all P〈0.05）. There were no significant differences between the MG and the SHPMG （all P〉0.05）. Conclusion： HPM can down-regulate the abnormally increased expressions of CRF and CRFR1 proteins and mRNAs in hypothalamus of the TNBS-induced experimental colitis rats, which is plausibly one of its action mechanisms in mitigating visceral pain and the pain-related emotions in the experimental colitis rats.
