Fexaramine Inhibits Receptor Activator of Nuclear Factor-κB Ligand-induced Osteoclast Formation via Nuclear Factor of Activated T Cells Signaling Pathways

BACKGROUND: Osteoclasts are bone resorbing cells and are responsible for bone erosion in diseases as diverse as osteoporosis, periodontitis, and rheumatoid arthritis. Fexaramine has been developed as an agonist for the farnesoid X receptor (FXR). This study investigated the effects of fexaramine on receptor activator of nuclear factor (NF)-κB ligand (RANKL)-induced osteoclast formation and signaling pathways. METHODS: Osteoclasts were formed by culturing mouse bone marrow-derived macrophages (BMMs) with macrophage colony-stimulating factor (M-CSF) and RANKL. Bone resorption assays were performed using dentine slices. The mRNA expression level was analyzed by real-time polymerase chain reaction. Western blotting assays were conducted to detect the expression or activation level of proteins. Lipopolysaccharide-induced osteoclast formation was performed using a mouse calvarial model. RESULTS: Fexaramine inhibited RANKL-induced osteoclast formation, without cytotoxicity. Furthermore, fexaramine diminished the RANKL-stimulated bone resorption. Mechanistically, fexaramine blocked the RANKL-triggered p38, extracellular signal-regulated kinase, and glycogen synthase kinase 3β phosphorylation, resulting in suppressed expression of c-Fos and NF of activated T cells (NFATc1). Consistent with the in vitro anti-osteoclastogenic effect, fexaramine suppressed lipopolysaccharide-induced osteoclast formation in the calvarial model. CONCLUSIONS: The present data suggest that fexaramine has an inhibitory effect on osteoclast differentiation and function, via downregulation of NFATc1 signaling pathways. Thus, fexaramine could be useful for the treatment of bone diseases associated with excessive bone resorption.

Attenuation of RANKL-induced Osteoclast Formation via p38-mediated NFATc1 Signaling Pathways by Extract of Euphorbia Lathyris L

BACKGROUND: Osteoclasts are the only cell type capable of breaking down bone matrix, and its excessive activation is responsible for the development of bone-destructive diseases. Euphorbia lathyris L. (ELL) is an herbal plant that belongs to the Euphorbiaceae family. This study investigated the effects of the methanol extract of the aerial part of ELL on receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclast formation and signaling pathways. METHODS: Osteoclasts were formed by co-culturing mouse bone marrow with osteoblasts or by culturing mouse bone marrow-derived macrophages (BMMs) with macrophage colony-stimulating factor (M-CSF) and RANKL. Bone resorption assays were performed using dentine slices. The expression level of mRNA was analyzed by real-time polymerase chain reaction (PCR) or reverse transcription (RT)-PCR. Western blotting assays were performed to detect the expression or activation level of proteins. RESULTS: ELL inhibited RANKL-induced osteoclast formation without cytotoxicity. Furthermore, the RANKL-stimulated bone resorption was diminished by ELL. Mechanistically, ELL blocked the RANKL-triggered p38 mitogen-activated protein kinase (MAPK) phosphorylation, which resulted in the suppression of the expression of c-Fos and nuclear factor of activated T cells (NFATc1). In osteoblasts, ELL had little effect on the mRNA expression of RANKL and osteoprotegerin (OPG). CONCLUSIONS: The present data suggest that ELL has an inhibitory effect on osteoclast differentiation and function via downregulation of the p38/c-Fos/NFATc1 signaling pathways. Thus, ELL could be useful for the treatment of bone diseases associated with excessive bone resorption.

Recombinant mycobacterium tuberculosis heat shock protein 10 in human osteoclast differentiation / 中国组织工程研究

BACKGROUND:The mycobacterium tuberculosis heat shock protein 10 exerts effects on the osteoclasts by in vitro mouse cranium experiment, OBJECTIVE:To investigate the effect and mechanism of recombinant mycobacterium tuberculosis heat shock protein 10 (CPN10) on the differentiation of osteoclasts in the in vitro culture system that induces osteoclast differentiation. METHODHuman macrophage colony-stimulating factor-dependent adhesive blood mononuclear cells were divided into four groupreceptor activator for nuclear factor-κB ligand (RANKL)+CPN10 (1 mg/L), RANKL, CPN10 (1 mg/L), and negative control (complete culture medium). Monocytes were resuspended in a-MEM medium containing macrophage colony-stimulating factor, and were cultured in each group for 7, 14, 21 days. The morphology, quantity and bone resorption area of osteoclasts were examined by tartrate-resistant acid phosphatase (TRAP) staining. The expressions of NFATc1 and c-Fos gene and protein were also detected. RESULTS AND CONCLUSION:In negative control group, no TRAP-positive multinucleated osteoclasts generated, while in the other groups, TRAP-positive multinucleated osteoclasts differentiated and formed the lacunae in the smal bone grinding. The number of osteoclasts formation and resorption in CPN10 group were significantly lower than that in RANKL+CPN10 group. The expression of NFATc1 and c-Fos in the negative control group C was significantly lower than that of RANKL+CPN10 group and CPN10 group. However, CPN10 expressed NFATc1 and c-Fos protein, which was significantly lower than RANKL+CPN10 group. CPN10 is involved in the formation of osteoclasts, and the mechanism is related with the upregulation of NFATc1, c-Fos expression.

Effects of acute millimeter wave exposure on the expression of substance P and c-fos in rat spinal cord / 解放军医学杂志

Objective To observe the expression changes in substance P (SP) and c-fos in rat spinal cord after acute millimeter-wave (MMW) exposure, and explore the mechanism of thermal hyperalgesia at the spinal level. Methods The back skin of SD rats was exposed to 35 GHz MMW (40W/cm2) for 0s (control group), 30s, 1min, or 3min. The corresponding segment of the spinal cord was taken at 0min, 5min, 10min, 1h and 3h after MMW irradiation for total RNA and protein extraction. The expressions of SP and c-fos mRNA were measured by real-time RT-PCR, and the expression of c-fos protein was detected by Western blotting. Results No significant difference was found between the control group and irradiation groups in SP and c-fos mRNA expression in the corresponding segment of spinal cord after MMW irradiation for 30s. After MMW irradiation for 1min, the SP and c-fos mRNA expressions in the corresponding segment of spinal cord increased significantly at 10min time point, and then decreased to the level of control group. After MMW irradiation for 3min, the SP and c-fos mRNA expression in the corresponding segment of spinal cord increased significantly at 5min, 10min and 1h time points, and decreased to the level of control group at 3h. No significant change was found in c-fos protein expression in the corresponding segment of spinal cord after MMW irradiation for 30s and 1min. After MMW irradiation for 3min, the c-fos protein expression in the corresponding segment of spinal cord increased significantly at 5min and 10min time point, and then decreased to the level of control group. Conclusion The increase of SP expression in rat skin after MMW irradiation may be related to the increase of SP and c-fos expressions in the corresponding segment of the spinal cord induced by thermal pain stimulation.

Effect of propofol on expression of c-fos mRNA in hippocampus in rats with visceral pain / 中华麻醉学杂志

Objective To investigate the effect of propofol on the expression of c-fos mRNA in hippocampus in rats with visceral pain (VP).Methods Thirty male Sprague-Dawley rats,weighing 180-240 g,were randomly divided into 3 groups (n =10 each):VP group,propofol 7.5 mg/kg group (group P1) and propofol 75.0 mg/kg group (group P2).0.9％ normal saline was injected intravenously via the caudal vein in group VP.Propofol 7.5 and 75.0 mg/kg were injected intravenously via the caudal vein in groups P1 and P2,respectively.VP was produced by colorectal distension in anesthetized rats.The threshold of VP was assessed by the intra-balloon pressure which was limited to 100 mm Hg to avoid damage to intestine before and after administration.The rats were sacrificed after measurement of the pain threshold,their brains were removed and hippocampi were isolated for determination of the expression of c-fos mRNA by RT-PCR.Results Compared with group VP,the threshold of VP was significantly increased and the expression of c-fos mRNA in hippocampus was down-regulated in groups P1 and P2 (P ＜ 0.05).The threshold of VP was significantly higher and the expression of c-fos mRNA in hippocampus was lower in group P2 than in group P1 (P ＜ 0.05).Conclusion Propofol can reduce VP through down-regulating the expression of c-fos mRNA in hippocampus in rats.

Effect of ketamine anesthesia in early pregnancy on the c-fos mRNA and c-jun mRNA expression in offsprings of rats / 中华麻醉学杂志

Objective To investigate the effect of ketamine anesthesia in the early pregnancy on the c-fos mRNA and c-jun mRNA expression in the offsprings of rats. Methods Thirty pregnant SD rats at 5-13 days of gestation were randomly divided into control group and ketamine group (n = 15 each). Ketamine 20 mg/kg was injected intravenously through tail vein followed by 2 h infusion at a rate of 130 mg·kg-1 ·h-1 in ketmine group.While the equal volume of normal saline was given instead of ketamine in control group. The learning and memory function of the offsprings were tested by Morris water maze test on postnatal day 20 and 30. The hippocampal tissues were taken to detect the expression of c-fos mRNA and c-jun mRNA and to observe the ultrastructure. Results Compared with group C, the escape latency was significantly prolonged at 2 days during the test which was performed on postnatal day 30, but there was no significant difference in the expression of c-fos mRNA and c-jun mRNA on postnatal day 20 and 30 and in the indices mentioned above on postnatal day 20 in ketamine group (P ＞0.05). The damage to hippocampal neurons happened in ketamine group. Conclusion The mechanism by which ketamine anesthesia in the early pregnancy inhibits the cognitive function of the offsprings is related to the hippocampal neuron damage, but not related to the expression of c-fos mRNA and c-jun mRNA in hippocampus.

Effects of rizatriptan on cortical spreading depression and c-Fos expression within periaqueductal grey / 中华神经科杂志

Objective To determine whether rizatriptan has an effect on cortical spreading depression (CSD) and c-Fos expression within periaqueductal grey (PAG) induced by CSD in rats. Methods The experimental SD rats were randomly divided into group A injected with KCl, group B KCl plus rizatriptan and group C NaCL The number and amplitude of CSD were recorded after KCl or NaCl injection. C-Fos positive neurons of different layer were identified by the immunohistochemical technique 2 hours after the first injection of KCl or NaCl. Results There was no CSD in group C. The number of CSD in group A ( 10.70±3.23 ) was significantly more than that in group B (6.10±2.56, t = - 3.528, P < 0.01 ). The amplitude of CSD in group A ( 17.33 (95% CI 11.45--23.11 ) mV) was significantly greater than that in group B (11.82 (95%CI 9.24--14.70) mV, Z= -4.360, P< 0.01). There were more cFos-like immnoreactive neurons in every layer in group A than in group C (P < 0.01 ) and in group B (P < 0.05 ). Conclusion Rizatriptan has an inhibitory effect on CSD, which might induce the headache through exciting the neurons in PAG.

Expression of apoptosis related gene in rats with diffuse brain injury combination secondary injury / 中国基层医药

Objective To explore the expression of c-fos,bcl-2,bcl-xL gene and apoptosis in rats with diffuse brain injury combination secondary injury(SBI).Methods 40 rats were randomly divided into the normal control group 10,10 cerebral ischemia,head injury group 10 and group 10 SBI to immunohistochemical method to detect brain cells in the c-fos,bcl-2,bcl-xL expression and in-situ to apoptosis(TUNEL).Results The expression of brain injury group and SBI group of cortex area c-fos gene (23.6±9.4),(37.1±5.5)positive cells/entries/H was significantly higher than that of cerebral ischemia group (5.6±1.4) positive cells/entries/H(t= 3.458,t = 3.648, all P<0.01) ;the expression of brain injury group and SBI group cortex area bcl-2 gene (18.2±4.6) ,(15.6±3.7)positive cells/entries/H lower than that brain blood group(23.6±4.3)positive cells/entries/H(t=2.345, t=2.447 ,all P <0.05) ;the expression of bcl-xL gene changes llttle;the apoptosis SBI group cortex area (36.6±5.3)cells/0.1mm2 higher than the brain injury group (21.6±4.4) cells/0.lmm2 (t = 2.378 ,P < 0.05 );the apoptosis and level of bcl-2 expression showed a negative correlation(r = -0.857 ,P <0.01 ).Conclusion The expression of c-fos,bcl-2,bcl-xL were increased with closely related to apoptosis in rats with diffuse brain injury combination secondary injury.

Inhibitory Action of Antisense c-jun oligodeoxynucleotides on Ultraviolet B Induced Fibroblast Damage / 中华皮肤科杂志

Objective To explore the inhibitory action of antisense c-jun oligodeoxynucleotides(ODN) on matrix metalloproteinase(MMP) expression of fibroblasts induced by ultraviolet B (UVB). Methods The c-jun and c-fos protein expression induced by UVB were measured by Western blot. The mRNA expression of c-jun was determined by reverse transcriptase polymerase chain reaction (RT-PCR) after transfection with c-jun antisense ODN. The pro-MMP-1 and MMP-3 synthesis of fibroblasts was measured by ELISA after treatment with UVB and antisense c-jun ODN. Results The UVB-induced c-jun protein expression of fibroblasts increased to 1.8, 2.6, 3.3 times compared with that of non-irradiated controls,while there was no significant change in c-fos protein expression. The pro-MMP-1 and MMP-3 synthesis induced by UVB irradiation increased obviously. After transfection with different concentrations of c-jun antisense ODN, the UVB-induced c-jun mRNA expression could be significantly inhibited(P

Interference effects of tetramethylpyrazine on the proliferation of vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ / 解放军医学杂志

Objective To study the interference effects of tetramethylpyrazine (TMP) on calcinuerin (CaN), c-fos and the nuclear antigen of proliferating cells in the proliferation of vascular smooth muscle cells (VSMCs) treated by angiotensinⅡ(Ang Ⅱ). Methods A cell proliferating model of VSMCs induced by Ang Ⅱ was established; the effects of TMP on CaN was detected by enzyme reaction phosphorus measurement; the effects of TMP on c-fos gene and PCNA expression were observed by immunocytochemical staining and image analysis technique (A value). Results The rats’ aortic smooth muscle cells were cultured in vitro successfully. CaN activities, cell proliferation activity and the expression levels of c-fos and PCNA increased significantly in VSMCs proliferation induced by Ang Ⅱ (P

Effects of acupuncture at Tsusanli (He-Sea Point) on expression of FOS and GFAP in medullary visceral zone of rats under stress / 解放军医学杂志

Objective To study the expression of Fos gene and GFAP in medullary visceral zone (MVZ) of rats under stress with their gastric mucosa protected by acupuncture. Methods 24 male SD rats were randomly divided into 3 groups: control, stress and acupuncture group, with 8 for each group. Immunohistochemical methods were employed to detect the expression of Fos and GFAP in the nucleus of solitary tract (NTS) and dorsal motor nucleus of vagus (DMV) of medullary visceral zone. The extent of gastric mucosal damage was evaluated by ulcer index (UI). Results Fos-positive neurons and GFAP-positive stellar microgliocytes were found mainly in NTS and DMV of the medullary visceral zone. Compared with control group, the expression of Fos and GFAP was greatly up-regulated in the stress group, and UI increased obviously (P

Relationship Between the Expression of c-fos Gene and Delayed Neuronal Death in the Neonatal Hippocampus Following Hypoxia-ischamia Insult / 中华围产医学杂志

Objective To explore the relationship between the expression of c fos gene and delayed neuron death in perinatal hypoxia ischamia encephalopathy (HIE). Methods Terminal deo xynuleotidyl transferase mediated dUTP nick end labeling (TUNEL), immuno histochemistry andreverse transcription PCR(RT PCR) were used to detect the apoptosis and transcription and translation of c fos gene in the hippocampus following HIE. Results Positive signal of apoptosis from CA1 to CA4 sections were observed in experimental hippocampi. Marked signal appeared earlier and longer in CA1 section than in the other sections[CA1: 1 h: (14.6?2.3),24 h: (51?6),72 h: (17.4?0.3);CA4:1 h :1.3?1.6),24 h: (47?8),72 h: (21.6?0.6) apoptosis cell number/ mm 2 ],compared with the sham group of CA1 section P

The expression of c-fos gene following cerebral hypoxia-ischemia in the neonatal rat / 中国病理生理杂志

AIM: To investigate the expression of c-fos in the neonatal rats' brains following hypoxia-ischemia.METHODS: RT-PCR and immunohistochemistry were used to detect the transcription and translation of c-fos gene in the cortex and hippocampus following hypoxia-ischemia.RESULTS: The expression of c-fos mRNA and protein were induced in the cortex and hippocampus at the early stage following hypoxia-ischemia(P

Fos expression and co-localization of 5-HT with Fos in periaqueductal grey in migraine model of rat / 中华神经科杂志

Objective To investigate the role of PAG in migraine Methods A transformed superior sinus stimulation migraine model was applied to observe Fos expression and co localization of 5 HT with Fos in PAG following the stimulation of dual matter adjacent to the superior sagittal sinus in rats Results 5 HT immunoreactive neurons, densely located in ventrolateral and ventromedial sectors, were found throughout the PAG Fos like immunoreactivity neurons were obviously increased in stimulated group than in unstimulated and control groups The double labelled neurons accounted for 15 0%?2 8% of 5 HT immunoreactive neurons and 24 0%?4 3% of Fos like immunoreactivity The double labelled population mainly distributed in ventrolateral sector Conclusions It suggests that neurons in PAG, especially in ventrolateral sector in caudal segment, might be excited in migraine model of rat The excited cells and 5 HTnic neurons might have a similar distribution pattern in PAG, with part of excited cells being 5 HTnic neurons Other types of excited neurons and the subtype of 5 HT receptor were not included in the experiment, and would be studied further as to getting a better understanding of the pathophysiology of migraine

Effects of propofol on c-fos mRNA expression in the different regions of brain following stress in rats / 中华麻醉学杂志

Objective To determine the effects of propofol on c fos gene expression in the different brain regions following stress in rats Methods Twenty one male Wistar rats (12 18 weeks) weighing 260 300g were randomly divided into three groups of seven animals each: control group(C); electrical stimulation group(S) and propofol group(P) The animals were anesthetized with pentobarbital sodium 40mg?kg -1 Normal saline 2ml (group C and S) or propofol 10mg?kg -1 (group P) was injected intraperitoneally (ip) 5 min after ip injection hindpaw of the animals in group S and P was electrically stimulated with 2 mA direct current (1 s every 30 s) for 15 min 30 min after electrical stimulation the animals were decapitated and brain was immediately removed on -20℃ ice plate and kept in -70℃ liquid nitrogen for determination of c fos mRNA expression in cerebral cortex, hypothalamus and hippocampus At the same time 4 ml of blood was collected from trunk for determination of ACTH and cortisol concentrations by immunoradiometric assay Results Plasma ACTH and cortisol levels and c fos mRNA expression in cerebral cortex, hypothalamus and hippocampus increased significantly in group S as compared with those in group C (P0 05).Conclusions The c fos gene is involved in molecular modulation of stress responses Propofol produces different effects on c fos gene expression in different brain regions

Effects of propofol on the anoxic response of primary cultured hippocampal neurons / 中华麻醉学杂志

Objective To evaluate the effects of different concentration of propofol on the anoxic response of primary cultured hippocampal neurons Methods Newborn (

Effect of propofol on fos expression in spinal cord induced by intraplantar injection of formalin / 中华麻醉学杂志

Objective To investigate the effect of propofol on the response of spinal cord to pain stimulation induced by intraplantar injection of formalin Methods Thirty SD rats of both sexes weighing 200 250g were randomly divided into six groups Pain stimulation was produced by subcutaneous injection of formalin (2 5%100?l) into the plantar region of unilateral front paw Group F received intraplantar injection of formalin only (n=6);group FP received intraperitoneal propofol 100?g?kg -1 10min after formalin injection (n=6); group PF received formalin injection 10min after intraperitoneal propofol 100?g?kg -1 (n=6); group P received intraperitoneal propofol 100?g?kg -1 only; group FS received intraperitoneal normal saline 10ml?kg -1 10min after formalin injection; and group S received intraperitoneal normal saline 10ml?kg -1 only 1 h after last injection (intraperitoneal or intraplantar) the animals were anesthetized and cervical spinal cord (where sensory nerves from front paw enter) was removed and sliced and examined for fos expression in the spinal cord using fos immunohistochemistry technique Results After formalin injection large numbers of fos like immunoreactive neurons (FLINs) were found in the ipsilateral dorsal horn Most of FLINs were confined to the medial part of outer area of laminal I and II Intraperitoneal propofol injected either before or after formalin stimulation significantly suppressed fos expression in all laminal (P

Effects of verapamil on hyperalgesia and spinal cord dorsal horn c-fos expression after trauma in rats / 中华麻醉学杂志

Objective To investigate the effects of verapamil on hyperalgesia and spinal cord dorsal horn c fos expression after trauma in rats Methods Forty Wistar rats weighing 160 220g were randomly divided into five groups with eight animals in each group Traumatic pain model was made by amputation of right hind limb 0 5cm below ankle joint In group A animals received intraperitoneal verapamil 10mg/kg after trauma; in group B fentanyl 30 ?g/kg; in group C verapamil 10mg/kg plus fentanyl 30?g/kg; in group D normal saline; in group E animals received neither trauma nor verapamil or fentanyl The pain threshold of the skin surrounding the site of trauma was measured at 15 min, 30 min,1h and 2h after trauma and the interval between the radiation heat applied to the skin over ankle and the withdrawal of the hind limb was taken as pain threshold Fos like immunoreactive (FLI) neurons were determined in 1 2 luminal of ipsilateral lumber spinal cord dorsal horn with FOS immunohistochemistry technique 2h after trauma Results The pain threshold of the skin surrounding trauma site decreased significantly at 15 min, 30 min, 1h and 2h after trauma (P

Effects of ketamine on c-fos gene expression during global myocardial ischemia-reperfusion in rat / 中华麻醉学杂志

Objective To detemine the effects of ketamine on c-fos gene expression during global myocardial ischemia-reperfusionMethods Forty Wistar rats were divided into 5 groups: group C as control; group CR as ischemia-reperfusion control and group KH,KM and KL treated with ketamine 1?10 -3,1?10 -4 and 1?10 -5mol/L respectively prior to ischemia-reperfusion Total cellular RNA of myocardium was extracted RT-PCR technique was applied to determining cDNA amplification products ?-actin mRNA served as an internal control Densities of DNA bands were quantified using computer image analysis systemResults As compared with the values of group C, c-fos mRNA levels were increased in group CR,KH,KM and KL(P005)Conclusions C-fos gene may involves in molecular modulation of myocardial ischemia-reperfusion injury and myocardial protection Ketamine can effectively depress the expression of c-fos gene in myocardium, the middle and the low concentrations of ketamine are more effective than the high concentrations

Effect of propofol on C-fos gene expression and neuronal damage induced by ketamine in rat cerebral cortex / 中华麻醉学杂志

Objective To investigate the effect of propofol on C-fos expression and glutamate concentration in rat cerebral cortex induced by ketamine. Methods Twenty-eight male Wistar rats weighing 260-280 g were randomly divided into four groups of seven animals: group 1 received normal saline intraperitoneally (ip) (group NS); group 2 received NS + ketamine 100mg?kg-1 ip (group K); group 3 received propofol 100 mg?kg-1 + ketamine 100mg?kg-1 ip (group PK); group 4 received diazepam 10mg?kg-1 + ketamine 100 mg?kg-1 ip (group DK). The interval between the two intraperitoneal injections was 5 min in each group. The animals were decapitated 30 min after ip injection. C-fos mRNA expression was detected by RT-PCR method and fos protein expression by immuno-histochemical technique. Another forty Wistar rats were randomly divided into 4 groups of 10 animals as was described above. Two hours after ip injection, five animals in each group were decapitated for microscopic examination and the other five animals for determination of water and glutamate content of cerebral cortex.Results C-fos mRNA expression increased at 30 min after intraperitoneal ketamine. Ketamine induced significant increase in Fos protein expression, and glutamate and water content in cerebral cortex 2 h after ip injection. Propofol and diazepam inhibited the increases induced by ketamine ( P