Post-stroke depression is associated with reduced expression of brain-derived neurotrophic factor （BDNF）. In this study, we evaluated whether BDNF overexpression affects depression-like behavior in a rat model of po...Post-stroke depression is associated with reduced expression of brain-derived neurotrophic factor （BDNF）. In this study, we evaluated whether BDNF overexpression affects depression-like behavior in a rat model of post-stroke depression. The middle cerebral artery was occluded to produce a model of focal cerebral ischemia. These rats were then subjected to isolation-housing combined with chronic unpredictable mild stress to generate a model of post-stroke depression. A BDNF gene lentiviral vector was injected into the hippocampus. At 7 days after injection, western blot assay and real-time quantitative PCR revealed that BDNF expression in the hippo- campus was increased in depressive rats injected with BDNF lentivirus compared with depressive rats injected with control vector. Furthermore, sucrose solution consumption was higher, and horizontal and vertical movement scores were increased in the open field test in these rats as well. These findings suggest that BDNF overexpression in the hippocampus of post-stroke depressive rats alleviates depression-like behaviors.展开更多
Acupuncture is a traditional therapeutic technique in Oriental medicine,which has a long history of 4000 years. As a modern version of hand acupuncture,electroacupuncture (EA) can provide a valid analgesic effect and ...Acupuncture is a traditional therapeutic technique in Oriental medicine,which has a long history of 4000 years. As a modern version of hand acupuncture,electroacupuncture (EA) can provide a valid analgesic effect and has little physiological interference. It was successfully used to ameliorate pain not only in varieties of painful diseases,but in various operations,such as cesarean section,gastrectomy,enterectomy and castration,in animals during the 1970s. Since then,analgesia-regulating mechanism of EA has been extensively investigated. Previous studies found that electroacupuncture analgesia (EAA) was involved in modulations of neurotransmitters or neuromodulators in the central nerve system (CNS),and most early studies focused on the role of neurotransmitters such as serotonin,noradrenaline, dopamine and acetylcholine. Later,it was certified that some endogenous opioid peptides (EOPs),mainly including enkephalin,β-endorphin and dynorphin,played a more important role in EAA. EA of different frequencies can promote the release of different EOPs in the CNS. Studies showed that EAA induced by 2 Hz (low frequency) was mediated by the release of met-enkephalin (M-ENK) and β-endorphin (β-EP),while EAA by 100 Hz (high frequency) was mediated by the release of dynorphin-A (DYN-A) in the CNS in rats. Although these results in rats above are extrapolated to give reasonable explanations for acupuncture analgesia phenomenon and its treatment of related diseases in human,there are still some unknown mechanisms to be investigated. It has been proved that analgesia induced by EA varies in animal species. In order to quantitatively estimate the degree of acupuncture-induced analgesia,some researchers used an anesthetic to ensure a complete analgesia and to assess the reduction of the amount of the anesthetics consumed in the EA plus anesthetic group as compared to the anesthetic group without acupuncture. Studies showed EA in combination with anesthetics resulted in the reduction of the dosage of the anesthetics in human,rat and goat by 45%-55%,50%-60% and over 75%,respectively. It is clear that the analgesic effect induced by EA in goats (ruminants) is superior to that in rats or human. Therefore,the modalities of EOP release elicited by different frequencies in ruminants could be different from those in rats. In the present study,goats were stimulated with EA of different frequencies to determine the analgesic efficacy and the release levels of M-ENK,β-EP and DYN-A in the CNS in order to probe into the mechanisms of EA-induced analgesia in ruminants. Goats were stimulated by electroacupuncture of different frequencies: 0,2,40,60,80 or 100 Hz at a set of Baihui,Santai,Ergen,and Sanyangluo points for 30 min. The pain threshold was measured using the method of potassium iontophoresis. The levels of met-enkephalin,β-endorphin and dynorphin-A were determined with SABC immunohistochemisty. The results showed that 60 Hz increased pain threshold by 91%; its increasing rate was higher (P < 0. 01) than that by any other frequency used. The three EOPs were distributed in most analgesia-related nuclei and areas in the CNS. 2 Hz and 100 Hz induced met-enkephalin immunoactivities to increase (P < 0. 05) in nucleus accumbens,septal area,caudate nucleus,amygdala,paraventricular nucleus of hypothalamus,periaqueductal gray,dorsal raphe nucleus and locus ceruleus. These two frequencies elicited β-endorphin immunoactivities to increase (P < 0. 05) in nucleus accumbens,septal area,supraoptic nucleus,ventromedial nucleus of hypothalamus,periaqueductal gray,dorsal raphe nucleus,locus ceruleus,solitary nucleus,amygdale. 60 Hz increased (P < 0. 05) the release of met-enkephalin or β-endorphin in the nuclei and areas mentioned above,and habenular nucleus,substantia nigra,parabrachial nucleus and nucleus raphe magnus. High frequencies induced dynorphin-A immunoactivities to increase (P < 0. 05) in spinal cord dorsal horn and most analgesia-related nuclei in the CNS. It suggested that 60 Hz induced the simultaneous release of the three EOPs in extensive analgesia-related nuclei and areas of the CNS,which may be contributive to optimal analgesic effects and species variation.展开更多
Quinolones (QNs) are widely used for their broad antibacterial spectrum and good antibacterial activities,desirable pharmacokinetic characteristics and few cross reactions with other therapeutic agents. However,QNs ha...Quinolones (QNs) are widely used for their broad antibacterial spectrum and good antibacterial activities,desirable pharmacokinetic characteristics and few cross reactions with other therapeutic agents. However,QNs have adverse effects including chondrotoxicity in juvenile animals,so the use of QNs is contraindicated in children and adolescents. In regarding to the chondrotoxicity of QNs,numerous studies have been done. The current hypothesis suggests that QNs compete with the β1 integrin receptors residing on chondrocyte surface for extracellular Mg ions,which leads to alternation in β1 integrin expression,or function and eventually results in chondrocyte death. Stupack et al (2001)demonstrated that caspase-8 could be recruited to unligated integrins in adherent cells and further initiate apoptosis in a death receptor-independent manner. Sheng et al (2008) found that ofloxacin induced rabbit's chondrocyte apoptosis by causing disturbance of β1 integrin functions and subsequently through caspase-8-dependent mitochondrial pathway.Apoptosis could be initiated through the stimulation of death receptors and through an intrinsic pathway from mitochondria. Santangelo and Bertone (2011) and Wu et al (2011) found that TNFα could be expressed in human primary condrocytes inducing by interleukin-1beta (IL-1) or lipopolysaccharide (LPS). However,to date there have not been sufficient results to support that signaling from the death receptors was involved in QNs-induced chondrocyte apoptosis.In addition,dilated cisternae of rough endoplasmic reticulum (ER) have been noticed in QNs-induced arthropathy.ER can participate in the initiation of apoptosis. Varieties of harmful cellular stimuli could lead to ERs (ER stress). Elevated ERs results in cellular apoptosis. But whether ERs mediates apoptosis in QNs-treated chondrocytes is not clear yet.In this study,we chose two QNs agents and chondrocytes were treated with ofloxacin and marbofloxacin at final concentrations of 20 μg / mL,50 μg / mL and 100 μg / mL respectively in vitro for 2 h,8 h and 24 h. Cell survival rate,cell apoptosis rate and death receptor pathway factors TNFα (intracellular tumor necrosis factor-alpha),TNFR1 (TNF receptor-1),TRADD (TNF receptor 1 associated via death domain),FADD (Fas-associated protein with death domain),caspase-8 and ERs mediated apoptosis factors caspase-12,GADD153 (CHOP or DDIT3),GRP78 (Bip),calpain,and anti-apoptosis factors Bcl-2 (B-cell lymphoma 2),NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) gene expression levels were measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) analysis to determine the dose-response relationship. We further silenced the expression of TNFR1 successfully by transferring TNFR1-siRNA to chondrocytes to confirm whether TNFα / TNFR1 signaling pathways are involved ofloxacin and marbofloxacin-induced apoptosis. Furthermore,expression of death receptor pathway representative proteins TNFα / TNFR1 and endocytoplasmic reticulum (ER) pathway representative protein caspase-12 were confirmed using Western Blot.We have found that ofloxacin and marbofloxacin could induce apoptosis of chondrocytes in a time-and dose-dependent fashion within 24 h. mRNA of TNFα,TNFR1,TRADD,FADD and caspase-8 (caspase-8 of ofloxacin treated group were at 24 h) were highly expressed at 8 h,and GADD153,GRP78,calpain and caspase-12 at 8 h or 24 h,and antiapoptosis factors NF-κB and Bcl-2 were also raised after 2 h,all in a dose-dependent fashion. Expression of caspase-8was downregulated after silenced TNFR1. TNFα and TNFR1 proteins were expressed at 8 h and caspase-12 proteins were expressed at 24 h. In addition,ofloxacin showed a higher toxicity.Our results indicate that death receptor pathway TNF / TNFR1 and ERs mediated apoptosis factors are involved in ofloxacin and marbofloxacin-induced apoptosis of in vitro cultured juvenile dog joint chondrocytes within 24 h.展开更多
基金supported by a grant from the Experimental Animal Science and Technology Project of Zhejiang Province in China,No.2012C37083
文摘Post-stroke depression is associated with reduced expression of brain-derived neurotrophic factor （BDNF）. In this study, we evaluated whether BDNF overexpression affects depression-like behavior in a rat model of post-stroke depression. The middle cerebral artery was occluded to produce a model of focal cerebral ischemia. These rats were then subjected to isolation-housing combined with chronic unpredictable mild stress to generate a model of post-stroke depression. A BDNF gene lentiviral vector was injected into the hippocampus. At 7 days after injection, western blot assay and real-time quantitative PCR revealed that BDNF expression in the hippo- campus was increased in depressive rats injected with BDNF lentivirus compared with depressive rats injected with control vector. Furthermore, sucrose solution consumption was higher, and horizontal and vertical movement scores were increased in the open field test in these rats as well. These findings suggest that BDNF overexpression in the hippocampus of post-stroke depressive rats alleviates depression-like behaviors.
文摘Acupuncture is a traditional therapeutic technique in Oriental medicine,which has a long history of 4000 years. As a modern version of hand acupuncture,electroacupuncture (EA) can provide a valid analgesic effect and has little physiological interference. It was successfully used to ameliorate pain not only in varieties of painful diseases,but in various operations,such as cesarean section,gastrectomy,enterectomy and castration,in animals during the 1970s. Since then,analgesia-regulating mechanism of EA has been extensively investigated. Previous studies found that electroacupuncture analgesia (EAA) was involved in modulations of neurotransmitters or neuromodulators in the central nerve system (CNS),and most early studies focused on the role of neurotransmitters such as serotonin,noradrenaline, dopamine and acetylcholine. Later,it was certified that some endogenous opioid peptides (EOPs),mainly including enkephalin,β-endorphin and dynorphin,played a more important role in EAA. EA of different frequencies can promote the release of different EOPs in the CNS. Studies showed that EAA induced by 2 Hz (low frequency) was mediated by the release of met-enkephalin (M-ENK) and β-endorphin (β-EP),while EAA by 100 Hz (high frequency) was mediated by the release of dynorphin-A (DYN-A) in the CNS in rats. Although these results in rats above are extrapolated to give reasonable explanations for acupuncture analgesia phenomenon and its treatment of related diseases in human,there are still some unknown mechanisms to be investigated. It has been proved that analgesia induced by EA varies in animal species. In order to quantitatively estimate the degree of acupuncture-induced analgesia,some researchers used an anesthetic to ensure a complete analgesia and to assess the reduction of the amount of the anesthetics consumed in the EA plus anesthetic group as compared to the anesthetic group without acupuncture. Studies showed EA in combination with anesthetics resulted in the reduction of the dosage of the anesthetics in human,rat and goat by 45%-55%,50%-60% and over 75%,respectively. It is clear that the analgesic effect induced by EA in goats (ruminants) is superior to that in rats or human. Therefore,the modalities of EOP release elicited by different frequencies in ruminants could be different from those in rats. In the present study,goats were stimulated with EA of different frequencies to determine the analgesic efficacy and the release levels of M-ENK,β-EP and DYN-A in the CNS in order to probe into the mechanisms of EA-induced analgesia in ruminants. Goats were stimulated by electroacupuncture of different frequencies: 0,2,40,60,80 or 100 Hz at a set of Baihui,Santai,Ergen,and Sanyangluo points for 30 min. The pain threshold was measured using the method of potassium iontophoresis. The levels of met-enkephalin,β-endorphin and dynorphin-A were determined with SABC immunohistochemisty. The results showed that 60 Hz increased pain threshold by 91%; its increasing rate was higher (P < 0. 01) than that by any other frequency used. The three EOPs were distributed in most analgesia-related nuclei and areas in the CNS. 2 Hz and 100 Hz induced met-enkephalin immunoactivities to increase (P < 0. 05) in nucleus accumbens,septal area,caudate nucleus,amygdala,paraventricular nucleus of hypothalamus,periaqueductal gray,dorsal raphe nucleus and locus ceruleus. These two frequencies elicited β-endorphin immunoactivities to increase (P < 0. 05) in nucleus accumbens,septal area,supraoptic nucleus,ventromedial nucleus of hypothalamus,periaqueductal gray,dorsal raphe nucleus,locus ceruleus,solitary nucleus,amygdale. 60 Hz increased (P < 0. 05) the release of met-enkephalin or β-endorphin in the nuclei and areas mentioned above,and habenular nucleus,substantia nigra,parabrachial nucleus and nucleus raphe magnus. High frequencies induced dynorphin-A immunoactivities to increase (P < 0. 05) in spinal cord dorsal horn and most analgesia-related nuclei in the CNS. It suggested that 60 Hz induced the simultaneous release of the three EOPs in extensive analgesia-related nuclei and areas of the CNS,which may be contributive to optimal analgesic effects and species variation.
文摘Quinolones (QNs) are widely used for their broad antibacterial spectrum and good antibacterial activities,desirable pharmacokinetic characteristics and few cross reactions with other therapeutic agents. However,QNs have adverse effects including chondrotoxicity in juvenile animals,so the use of QNs is contraindicated in children and adolescents. In regarding to the chondrotoxicity of QNs,numerous studies have been done. The current hypothesis suggests that QNs compete with the β1 integrin receptors residing on chondrocyte surface for extracellular Mg ions,which leads to alternation in β1 integrin expression,or function and eventually results in chondrocyte death. Stupack et al (2001)demonstrated that caspase-8 could be recruited to unligated integrins in adherent cells and further initiate apoptosis in a death receptor-independent manner. Sheng et al (2008) found that ofloxacin induced rabbit's chondrocyte apoptosis by causing disturbance of β1 integrin functions and subsequently through caspase-8-dependent mitochondrial pathway.Apoptosis could be initiated through the stimulation of death receptors and through an intrinsic pathway from mitochondria. Santangelo and Bertone (2011) and Wu et al (2011) found that TNFα could be expressed in human primary condrocytes inducing by interleukin-1beta (IL-1) or lipopolysaccharide (LPS). However,to date there have not been sufficient results to support that signaling from the death receptors was involved in QNs-induced chondrocyte apoptosis.In addition,dilated cisternae of rough endoplasmic reticulum (ER) have been noticed in QNs-induced arthropathy.ER can participate in the initiation of apoptosis. Varieties of harmful cellular stimuli could lead to ERs (ER stress). Elevated ERs results in cellular apoptosis. But whether ERs mediates apoptosis in QNs-treated chondrocytes is not clear yet.In this study,we chose two QNs agents and chondrocytes were treated with ofloxacin and marbofloxacin at final concentrations of 20 μg / mL,50 μg / mL and 100 μg / mL respectively in vitro for 2 h,8 h and 24 h. Cell survival rate,cell apoptosis rate and death receptor pathway factors TNFα (intracellular tumor necrosis factor-alpha),TNFR1 (TNF receptor-1),TRADD (TNF receptor 1 associated via death domain),FADD (Fas-associated protein with death domain),caspase-8 and ERs mediated apoptosis factors caspase-12,GADD153 (CHOP or DDIT3),GRP78 (Bip),calpain,and anti-apoptosis factors Bcl-2 (B-cell lymphoma 2),NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) gene expression levels were measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) analysis to determine the dose-response relationship. We further silenced the expression of TNFR1 successfully by transferring TNFR1-siRNA to chondrocytes to confirm whether TNFα / TNFR1 signaling pathways are involved ofloxacin and marbofloxacin-induced apoptosis. Furthermore,expression of death receptor pathway representative proteins TNFα / TNFR1 and endocytoplasmic reticulum (ER) pathway representative protein caspase-12 were confirmed using Western Blot.We have found that ofloxacin and marbofloxacin could induce apoptosis of chondrocytes in a time-and dose-dependent fashion within 24 h. mRNA of TNFα,TNFR1,TRADD,FADD and caspase-8 (caspase-8 of ofloxacin treated group were at 24 h) were highly expressed at 8 h,and GADD153,GRP78,calpain and caspase-12 at 8 h or 24 h,and antiapoptosis factors NF-κB and Bcl-2 were also raised after 2 h,all in a dose-dependent fashion. Expression of caspase-8was downregulated after silenced TNFR1. TNFα and TNFR1 proteins were expressed at 8 h and caspase-12 proteins were expressed at 24 h. In addition,ofloxacin showed a higher toxicity.Our results indicate that death receptor pathway TNF / TNFR1 and ERs mediated apoptosis factors are involved in ofloxacin and marbofloxacin-induced apoptosis of in vitro cultured juvenile dog joint chondrocytes within 24 h.
