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1.
Glycogen synthase kinase-3beta (GSK-3beta) is a serine/threonine kinase that plays a role in glycogen synthesis by inhibiting glycogen synthase (GS) through phosphorylation. We hypothesized that GSK-3beta by virtue of its role in glycogen synthesis through the inhibition of GS will play a role in the preparation of the endometrium for blastocyst implantation. Immunohistochemical (IHC) analysis and Western blot analysis (WBA) detected GSK-3beta in the endometrium, myometrium, Fallopian tube and ovary. WBA showed more than 5-fold higher endometrial expression of the phosphorylated GSK-3beta (pGSK-3beta) isoform (inactive) in the secretory phase as compared with the proliferative phase (P < 0.001), whereas no differences in total GSK-3beta expression were detected. IHC analysis confirmed the WBA and showed marked expression of pGSK-3beta predominantly in glandular epithelial cells in early and mid secretory endometrium with scant expression during the proliferative phase. In in vitro experiments using human endometrial-derived epithelial cell line (HES), progesterone did not alter total GSK mRNA or protein expression. However, progesterone induced a dose-dependent increase in the expression of pGSK-3beta, which could be blocked by RU486. Cyclic expression of GSK-3beta's active and inactive forms in the endometrium suggests that sex hormones regulate the expression of this enzyme. In vitro experiments demonstrate that progesterone through receptor-mediated mechanisms induces phosphorylation of endometrial GSK-3beta. 相似文献
2.
The activity of protein phosphatase-2A (PP-2A) is significantly suppressed in the brain of Alzheimer's disease (AD) patients, but the mechanism is not understood. Here, we found an in vivo association of glycogen synthase kinase 3β (GSK-3β) with inhibitor-2 of PP-2A (I 2PP-2A). The activation of GSK-3 resulted in accumulation of I 2PP-2A with concomitant suppression of PP-2A activity and increases of tau phosphorylation in HEK293, N2a and PC12 cells, while inhibition of GSK-3 caused decreases of I 2PP-2A with increased PP-2A activity and decreased tau phosphorylation. A positive correlation between GSK-3β and I 2PP-2A ( R = 0.9158) and a negative correlation between GSK-3β and PP-2A ( R = −0.9166) were detected. GSK-3 activation did not affect I 2PP-2A mRNA level, while it increased the mRNA level of a heterogeneous ribonucleoprotein A18 (hnRNP A18). The activation of GSK-3 increased the expression and the activity of proteasome system. It suggests that activation of GSK-3 inhibits PP-2A through up-regulation of I 2PP-2A with hnRNP A18-involved mechanism. 相似文献
4.
Occipital artery (OA) injections of 5-HT elicit pronounced reductions in heart rate and mean arterial blood pressure (MAP) in urethane-anesthetized rats by activation of vagal afferent cell bodies in the ipsilateral nodose ganglion. In contrast, internal carotid artery (ICA) and i.v. injections elicit similar cardiovascular responses by activation of peripheral vagal afferent terminals. The aim of this study was to examine the roles of 5-HT3 and 5-HT2 receptors in the 5-HT-induced activation of vagal afferent cell bodies and peripheral afferent terminals in urethane-anesthetized rats. OA, ICA and i.v. injections of 5-HT elicited dose-dependent reductions in heart rate and MAP that were virtually abolished after i.v. administration of the 5-HT3 receptor antagonists, MDL 7222 or ICS 205-930. The responses elicited by the OA injections of 5-HT were markedly diminished after i.v. injection of the 5-HT2 receptor antagonists, xylamidine or ketanserin, whereas the responses elicited by i.v. or ICA injections of 5-HT were not affected. The present findings suggest that (1) 5-HT3 and 5-HT2 receptor antagonists gain ready access to nodose ganglion cells upon i.v. administration, and (2) functional 5-HT3 and 5-HT2 receptors exist on the cell bodies of vagal afferent neurons mediating the cardiovascular responses elicited by OA injections of 5-HT. These findings also support a wealth of evidence that 5-HT3 receptors exist on the peripheral terminals of vagal afferents, and although they do not discount the possibility that 5-HT2 receptors exist on peripheral vagal afferent terminals, it appears that activation of these receptors does not have pronounced effects on 5-HT3 receptor activity on terminals that mediate the hemodynamic responses to 5-HT. 相似文献
5.
全世界约有1.7亿人感染丙型肝炎病毒(hepatitis C virus,HCV),占世界人口的3%,我国约有4千万HCV携带者,平均感染率为3.2%.尽管急性感染阶段症状轻微,但70%以上HCV感染者发展成慢性过程,可导致慢性活动性肝炎和肝硬化,甚至肝细胞癌,严重威胁着人类的健康[1].目前多采用干扰素-α(interferon-α,IFN-α)或者IFN-α联合病毒唑进行治疗,但IFN-α治疗有其局限性,存在着疗效差、易产生耐药性、价格昂贵、复发率高等缺点[2]. 相似文献
6.
Exposure of GH 3 rat anterior pituitary cells to cholera toxin for 2–4 h significantly increased the thyrotropin-releasing-hormone(TRH)-induced inhibition of the inwardly rectifying K + current studied in patchperforated voltage-clamped cells. On the other hand, the current reduction became almost totally irreversible after washout of the neuropeptide. Comparison of the effects elicited by the toxin with those of 8-(4-chlorophenylthio)-cAMP or forskolin plus isobutylmethylxanthine indicated that, although the irreversibility may be due, at least in part, to elevations of cAMP levels, the enhancement of the TRH-induced inhibition of the current is not mediated by the cyclic nucleotide. Only reductions on the inwardly rectifying K + current, but not those elicited by TRH on voltage-dependent Ca 2+ currents, were increased by the treatment with cholera toxin. In current-clamped cells showing similar rates of firing, the second phase of enhanced action-potential frequency induced by TRH was also significantly potentiated by cholera toxin. Measurements of [Ca 2+] i oscillations associated with electrical activity, using video imaging with fura-2-loaded cells, demonstrated that cholera toxin treatment causes a clear reduction of spontaneous [Ca 2+] i oscillations. However, this did not prevent the stimulatory effect of TRH on oscillations due to the action potentials. In cholera-toxin-treated cells, the steady-state, voltage dependence of inactivation of the inward rectifier was shifted by nearly 20 mV to more negative values. These data suggest that the inwardly rectifying K + current plays an important role in maintenance of the resting K + conductance in GH 3 cells. Furthermore, the TRH-induced reductions on this current may be an important factor contributing to the increased cell excitability promoted by the neuropeptide. 相似文献
7.
Background/aims: Connective tissue growth factor (CTGF) is a downstream mediator of transforming growth factor-beta 1 (TGF-β 1) and TGF-β 1-induced CTGF expression is regulated through SMAD and mitogen-activated protein kinase (MAPK) signaling pathways. However, little is known about the localization of CTGF and TGF-β 1 signaling cascades during incisor development. Therefore, we aimed to investigate the distribution pattern of TGF-β 1, CTGF, phosphorylated-SMAD2/3 (p-SMAD2/3), and phosphorylated-ERK1/2 (p-ERK1/2) in the developing mouse incisors. Materials and methods: ICR mice heads of embryonic (E) day 16.5, postnatal (PN) day 0.5 and PN3.5 were processed for immunohistochemistry. Results: From E16.5 to PN3.5, moderate to strong staining for TGF-β1 and CTGF was localized in stellate reticulum (SR), transit amplifying (TA) cells, outer enamel epithelium (OEE), preameloblasts (PA), preodontoblasts (PO), and dental papilla (DP). p-SMAD2/3 was weakly positive in SR and OEE at E16.5 and PN0.5 but was strongly positive in SR and OEE at PN3.5. Particularly, in the stem cell niche, p-SMAD2/3 was only localized in SR cells adjacent to OEE. There was no staining for p-SMAD2/3 in TA cells, PA and PO, although weak to moderate staining for p-SMAD2/3 was seen in DP. From E16.5 to PN3.5, p-ERK1/2 was negative in TA cells, OEE, PA and PO, whereas weak to moderate staining for p-ERK1/2 was observed in SR. DP was moderately stained for p-ERK1/2. Conclusions: TGF-β1 and CTGF show a similar expression, while p-SMAD2/3 and p-ERK1/2 exhibit differential distribution pattern, which indicates that CTGF and TGF-β1 signaling cascades might play a regulatory role in incisor development. 相似文献
8.
Neuromodulation with low-frequency stimulation (LFS), of brain structures other than epileptic foci, is effective in inhibiting seizures in animals and patients, whereas selection of targets for LFS requires further investigation. The hippocampal CA 3 subfield is a key site in the circuit of seizure generation and propagation. The present study aimed to illustrate the effects of LFS of the CA 3 region on seizure acquisition and generalization in the rat amygdaloid kindling model of epilepsy. We found that LFS (monophasic square-wave pulses, 1 Hz, 100 μA and 0.1 ms per pulse) of the CA 3 region significantly depressed the duration of epileptiform activity and seizure acquisition by retarding progression from focal to generalized seizures (GS). Moreover, GS duration was significantly shortened and its latency was significantly increased in the LFS group demonstrating an inhibition of the severity of GS and the spread of epileptiform activity. Furthermore, LFS prevented the decline of afterdischarge threshold (ADT) and elevated GS threshold indicating an inhibition of susceptibility to GS. These results suggest that LFS of the hippocampal CA 3 subfield is anti-epileptogenic and anti-ictogenic. Neuromodulation of CA 3 activity using LFS may be an alternative potential approach for temporal lobe epilepsy treatment. 相似文献
9.
Increases in solution pH from 6.5 to 7.0 to 7.5 at 0.1 M free Ca 2+ concentration had no effect on the isometric tension of barnacle myofibrillar bundles in relaxing solutions containing 0.1–0.16 mM BAPTA. Decreases in pH in the same range were also without effect. Under the same conditions CO 2-induced Ca 2+ release from the SR could be readily obtained by replacing the Cl –-containing relaxing solution with one containing HCO
3
–
and 100% CO 2 at the same pH. At a higher free Ca 2+ of 2.5 M, there was a contraction on increasing the pH of the Cl –-containing solution from 7.0 to 7.5. This reponse could be abolished by 1 mM procaine suggesting that it was due to Ca 2+ release from the SR. The protonophores monensin, gramicidin, CCCP and FCCP at concentrations of 10–100 M had no effect on resting tension at either free Ca 2+ concentration and did not inhibit the response to 100% CO 2. It is concluded that dissipation of a possible pH gradient across the SR membrane by protonophores does not release Ca 2+ from the SR of barnacle muscle. Since both CO 2 (by possibly lowering SR pH) and an increase in solution pH can release Ca 2+ at 2.5 M free Ca 2+, the existence of a Ca 2+ release channel which is opened by a change in the trans-SR pH gradient cannot be discounted. In a separate series of experiments, the CO 2-releasable Ca 2+ store was first depleted by exposure of bundles to 100% CO 2 in the presence of 1 mM BAPTA for 10 min and then partially reloaded by 15 s exposure to a free Ca 2+ of 0.6 M buffered by 2 mM EGTA (total) in the Cl – relaxing solution. The same level of reloading was obtained when the loading solution contained HCO
3
–/100%
CO 2; this result tends to discount inhibition of the Ca 2+ uptake pump as a possible mechanism for CO 2-induced Ca 2+ release. Loading at 2.6 M free Ca 2+ for 1 min resulted in almost complete recovery of the CO 2 response to its value before depletion of the store. 相似文献
10.
Extracellular amyloid beta (Abeta) that confers neurotoxicity and modulates synaptic plasticity and memory function has been central to the amyloid hypothesis of Alzheimer's disease (AD) pathology. Like many other misfolded proteins identified in neurodegenerative disorders, Abeta also accumulates inside the AD neurons. This intracellular Abeta affects a variety of cellular physiology from protein degradation, axonal transport, autophagy to apoptosis, further documenting the role of Abeta in AD. Therapeutics targeting intracellular Abeta could be effective treatment for AD. 相似文献
12.
Oxidative stress by exposure to H 2O 2 induces various types of cell death depending on cell type and conditions. We report herein on a study of the mechanisms underlying H 2O 2-induced cell death in C6 glioma cells. The findings show that H 2O 2 triggers a caspase-independent autophagic cell death in these cells. The findings also show that H 2O 2 induces the dephosphorylation of the mammalian target of rapamycin (mTOR) at Ser 2481 and the p70 ribosomal protein S6 kinase (p70S6K) at Thr389 in a Bcl-2/E1B 19 kDa interacting protein 3 (BNIP3)-dependent manner. BNIP3 has the capacity to inhibit mTOR activity and mTOR inhibition plays a role in autophagic induction. This suggests that BNIP3 may mediate H 2O 2-induced autophagic cell death through the suppression of mTOR. The findings show that the down-regulation of BNIP3 by BNIP3 siRNA prevents C6 cells from undergoing H 2O 2-induced autophagic cell death. Collectively, these results suggest that H 2O 2 induces autophagic cell death in C6 cells via the BNIP3-mediated suppression of the mTOR pathway. 相似文献
13.
Activated microglia produces inflammatory cytokines and nitric oxide (NO) that involved in neuronal injury and neurodegenerative diseases. We report herein, that H(2)O(2) intensifies the LPS-triggered expression of iNOS in the microglia cell line, BV-2, resulting in an enhancement in the production of NO. The NO production induced by a combination of LPS and H(2)O(2) was blocked by the addition of an anti-interferonβ (IFNβ) neutral antibody, suggesting that IFNβ levels are correlated with the LPS/H(2)O(2)-induced production of NO. However, although the expression of IFNβ was induced by H(2)O(2) treatment alone, neither the expression of iNOS mRNA nor the production of NO were induced. In addition, the expression of IFN receptor (IFNR) was induced by LPS but not by H(2)O(2). These data indicate that although H(2)O(2) alone cannot induce iNOS expression because of the insufficient expression of IFNR, in the presence of LPS, H(2)O(2) enhances iNOS expression via the expression of IFNβ. Our findings suggest that H(2)O(2) produced by activated microglia further enhances NO production in various inflammatory states. 相似文献
14.
Oxidative stress has been suggested as a mechanism contributing to neuronal death induced by hypoglycemia, and an early production of reactive species (RS) during the hypoglycemic episode has been observed. However, the sources of reactive oxygen (ROS) and nitrogen (RNS) species have not been fully identified. In the present study we have examined the contribution of various enzymatic pathways to RS production and neuronal death induced by glucose deprivation (GD) in hippocampal cultures. We have observed a rapid increase in RS during GD, which depends on the activation of NMDA and non-NMDA receptors and on the influx of calcium from the extracellular space. Accordingly, intracellular calcium concentration [Ca 2+] i progressively increases more than 30-fold during the GD period. It was observed that superoxide production through the activation of the calcium-dependent enzymes, phospholipase A 2 (cPLA 2) and xanthine oxidase (XaO), contributes to neuronal damage, while nitric oxide synthase (NOS) is apparently not involved. Inhibition of cPLA 2 decreased RS at early times of GD whereas inhibition of XaO diminished RS at more delayed times. The antioxidants trolox and ebselen also showed a protective effect against neuronal death and diminished RS generation. Inhibition of NADPH oxidase also contributed to the early generation of superoxide. Taking together, the present results suggest that the early activation of calcium-dependent ROS producing pathways is involved in neuronal death associated with glucose deprivation. 相似文献
15.
Metastasis, the major cause of cancer death, is a multistep process that requires interactions between cancer cells and stromal cells and between cancer cells and extracellular matrix. Molecular alterations of the extracellular matrix in the tumor microenvironment have a considerable impact on the metastatic process during tumorigenesis. Here we report that elevated expression of betaig-h3/TGFBI (transforming growth factor, beta-induced), an extracellular matrix protein secreted by colon cancer cells, is associated with high-grade human colon cancers. Ectopic expression of the betaig-h3 protein enhanced the aggressiveness and altered the metastatic properties of colon cancer cells in vivo. Inhibition of betaig-h3 expression dramatically reduced metastasis. Mechanistically, betaig-h3 appears to promote extravasation, a critical step in the metastatic dissemination of cancer cells, by inducing the dissociation of VE-cadherin junctions between endothelial cells via activation of the integrin alphavbeta5-Src signaling pathway. Thus, cancers associated with overexpression of betaig-h3 may have an increased metastatic potential, leading to poor prognosis in cancer patients. 相似文献
16.
We used western blotting to measure the quantity of glutamate and γ-aminobutyric acid (GABA) transporters proteins within
hippocampal tissue obtained from rats who had undergone epileptogenesis. Chronic seizures were induced by amygdalar injection
of FeCl 3. We found that the glial glutamate transporters GLAST and GLT-1 were down-regulated at 60 days after initiation of chronic
and recurrent seizures. However, the neuronal glutamate transporter EAAC-1 and the GABA transporter GAT-3 were increased.
We performed in vivo microdialysis in freely moving animals to estimate in vivo redox state. We found that the hippocampal
tissues were oxidized, resulting in even further impairment of glutamate transport. Our data show that epileptogenesis in
rats resulting in chronic and recurrent seizures is associated with collapse of glutamate regulation caused by both the molecular
down-regulation of glial glutamate transporters combined with the functional failure due to oxidation. 相似文献
17.
ATP is implicated in peripheral pain signaling by actions on P2X receptors, especially P2X(3) receptor. Cardiac primary afferents running in the sympathetic nerves are considered to be essential pathways for transmission of cardiac nociception to the central nervous system. Because little is known about P2X(3) involvement in cardiac nociception, this study observed the difference in P2X(3) localization and expression in stellate ganglia (SG) from naive rats and in a pathological model of myocardial ischemic injury induced by repeated subcutaneous isoprenaline injections. Distribution of P2X(3) and morphometry of neurons in SG were investigated by immunohistochemistry, Western blotting, in situ hybridization (ISH) and by sterological study. Diffuse cytoplasmic P2X(3) immunolabelling was observed by light microsocopy. No nuclear labeling was detected. The intensity of P2X(3) labeling in the experimental myocardial ischemic injury group was increased in relation to that of the control group. Numerical densities of stellate ganglion neurons in the experimental group were higher than those of the control group. By Western blotting and ISH, the signals of P2X(3) protein and its mRNA in the myocardial ischemic group were higher than those of the control group. The P2X(3) labeling intensity and the numerical density in SG of the experimental myocardial ischemic injury group were enhanced, suggesting the involvement of P2X(3) receptor for the transmission of pain after myocardial ischemic injury. 相似文献
18.
Somatic mutations of PIK3CA are found in 20% to 40% of invasive breast cancers. To investigate the frequency of PIK3CA mutations in the intraductal proliferative lesions of the breast, which are precursor lesions for invasive carcinoma, we analyzed 125 intraductal proliferative lesions and 108 invasive breast cancer tissues for PIK3CA mutations in this study. Target cells were precisely isolated using a laser capture microdissection (LCM) system. Genomic DNA was extracted with QIAmp DNA Micro Kit. PCR amplification was done for exons 9 and 20 of PIK3CA, where 90% of mutations clustered, and the products were directly sequenced. Forty-six missense mutations were identified in total, of which, 14 and 32 mutations clustered in exon 9 and exon 20, respectively. The most common mutations were E542K (6 cases) and E545K (8 cases) in exon 9, and H1047R (29 cases) in exon 20. A novel mutation, G3292T, was also found. Mutations were found less frequently in the ductal intraepithelial neoplasia 1B (DIN1B) and lower grade ductal proliferative lesions (3 of 68; 4.41%) than in ductal carcinoma in situ (14 of 57; 24.56%, P = 0.001, Chi-square test) or invasive carcinoma (29 of 108; 26.85%, P = 0.000, Chi-square test). However, there was no significant difference in the frequency of PIK3CA mutations between carcinoma in situ and invasive carcinoma ( P = 0.750, Chi-square test). PIK3CA mutations mostly began to develop at the stage from the DIN1B to the carcinoma in situ (DCIS), which is a late event of breast oncogenesis. PIK3CA-mutated tumors were more frequently found in ER-a positive, PR positive, and PTEN positive tumors ( P = 0.012, P = 0.004 and P = 0.004, respectively, Chi-square test). The frequency of PIK3CA gene mutation in ER+/PR+ (32/98, 32.65%) tumors was not significantly different from that in ER+/PR− (9/39, 23.08%), tested by the Chi-square test ( P = 0.269). There was no significant association between PIK3CA mutations and HER2 expression status ( P = 0.294, Chi-square test). 相似文献
19.
We examined the relationship between the personality trait of harm avoidance (HA) and the dopamine D 2/3 receptor availability in striatal subdivisions using high resolution positron emission tomography (PET) with [ 11C]raclopride. Twenty-one healthy subjects completed 3 T magnetic resonance imaging and high resolution PET scans with [ 11C]raclopride in order to measure D 2/3 receptor availability in subregions of the striatum. The D 2/3 receptor availability was obtained on the basis of the Logan graphical method. The Temperament and Character Inventory was used to measure the biogenetic temperament of HA. The analysis revealed that the HA score had significant negative correlations with D 2/3 receptor availability in the pre-commissural dorsal caudate and post-commissural putamen, suggesting that HA is associated with D 2/3 receptor availability in the associative and sensorimotor subdivisions of the striatum, which are mainly involved in cognition and locomotion. Further research is required to determine if pathological states have similar dopaminergic mechanisms in specific striatal subdivisions. 相似文献
20.
Background: Enhanced gallbladder concentrating function is an important factor for the pathogenesis of cholesterol gallstone disease (CGD), but the mechanism is unknown. Potential candidates for regulation of gallbladder ion absorption are suggested to be Na +/H + exchanger isoform 3 (NHE 3). In this study, we investigated the expression and subcellular localization of NHE 3 in both acalculous and calculous human gallbladders. Methods: Adult human gallbladder tissue was obtained from 23 patients (7 men, 16 women) who had undergone cholecystectomy. The patients were divided into two groups: Group A (acalculous group) and Group B (calculous group). Gene expression of NHE 3 was quantitatively estimated by real-time PCR. Protein expression was studied by Western blotting assays. Furthermore, expression of immunoreactive NHE 3 was investigated by immunohistochemistry. Results: There was no significant difference in the NHE 3 mRNA expression between calculous and acalculous human gallbladders. NHE 3 protein expression in gallbladders from patients with cholelithiasis is increased compared to those without gallstones. Immunohistochemistry studies prove that NHE 3 is located both on the apical plasma membrane and in the intracellular pool in human GBECs. Conclusions: NHE 3 may play a role in the pathogenesis of human CGD. Additional studies are required to further delineate the underlying mechanisms. 相似文献
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