盐诱导激酶(salt-inducible kinases,SIKs)属于腺苷酸活化激酶(AMP-activated protein kinase,AMPK)家族,主要调控环磷酸苷效应元件结合蛋白(cAMP-response element binding protein,CREB)转录共激活因子(CREB-regulated transcription ...盐诱导激酶(salt-inducible kinases,SIKs)属于腺苷酸活化激酶(AMP-activated protein kinase,AMPK)家族,主要调控环磷酸苷效应元件结合蛋白(cAMP-response element binding protein,CREB)转录共激活因子(CREB-regulated transcription coactivators,CRTCs)的胞核-胞质分布,对CRTC-CREB复合物的合成起到调节作用,从而间接影响CREB目的基因的转录与表达,影响多种生理过程,例如能量代谢、细胞周期进程和细胞凋亡。过去对SIKs的相关研究主要集中在外周系统疾病,包括肿瘤、高血压和糖异生等。近年来越来越多的研究开始探索SIKs和中枢神经系统疾病的关联性,如抑郁症、睡眠障碍、癫痫和阿尔茨海默病等都存在SIKs水平异常的情况,这提示SIKs信号失调参与这些疾病的病理过程,SIKs具有成为此类疾病新治疗靶点的潜力。本文就SIKs在抑郁症、睡眠障碍及其他神经系统疾病中的作用和调控机制进行综述。展开更多
探讨柴金解郁安神片调控前扣带皮层(ACC)-腹侧海马(vHPC)谷氨酸能神经环路异常改善抑郁症大鼠腹侧海马神经元突触重塑的分子机制。首先运用化学遗传将谷氨酸能腺相关病毒(AAV)定位注射至大鼠ACC脑区,并通过慢性温和不可预知性应激(CUMS...探讨柴金解郁安神片调控前扣带皮层(ACC)-腹侧海马(vHPC)谷氨酸能神经环路异常改善抑郁症大鼠腹侧海马神经元突触重塑的分子机制。首先运用化学遗传将谷氨酸能腺相关病毒(AAV)定位注射至大鼠ACC脑区,并通过慢性温和不可预知性应激(CUMS)联合孤笼饲养复制大鼠抑郁模型,实验设正常组、模型组、AAV空载组、AAV病毒组、AAV病毒+糖皮质激素受体(GR)阻断剂组、AAV病毒+趋化因子受体1(CX3CR1)阻断剂组、AAV病毒+柴金解郁安神片组,采用水迷宫(Morris water maze)、旷场(open-field)和强迫游泳(forced-swimming)实验联合动物行为分析系统评估大鼠抑郁样行为;苏木素-伊红(HE)染色检测大鼠ACC及vHPC脑区神经元形态结构变化;免疫荧光及核磷酸蛋白(c-Fos)检测大鼠ACC-vHPC谷氨酸能神经环路激活情况;高尔基染色和透射电镜检测大鼠vHPC神经元树突、树突棘及突触亚微结构变化;免疫荧光、Western blot分别检测大鼠vHPC谷氨酸能神经元细胞内突触重塑相关蛋白谷氨酸受体2A(GRIN2A)、谷氨酸受体2B(GRIN2B)、Ca^(2+)/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)、丝裂原激活蛋白激酶激活蛋白激酶2(MK2)、丝切蛋白(cofilin)表达水平。结果表明,谷氨酸能AAV病毒激活后模型组大鼠抑郁样行为表型、ACC及vHPC神经元形态结构、突触超微结构损伤更加加重,而GR、CX3CR1阻断剂均能不同程度逆转其异常改变,提示ACC脑区内胶质细胞GR/CX3CR1双信号介导的ACC-vHPC谷氨酸能神经环路异常激活可能与抑郁的发生发展密切相关。有趣的是,柴金解郁安神片也能显著抑制AAV病毒诱导的ACC-vHPC神经环路激活及Glu含量异常升高,同时有效逆转模型组大鼠进一步加重的抑郁样行为和vHPC谷氨酸能神经元突触重塑,并揭示其改善腹侧海马神经元突触损伤的分子机制可能与调控突触重塑相关信号NR/CaMKⅡ、MK2/cofilin有关。综上,该文证实了柴金解郁安神片能有效调控ACC-vHPC谷氨酸能神经环路异常进而改善抑郁症大鼠腹侧海马谷氨酸能神经元突触重塑,其分子机制可能与调节突触相关NR/CaMKⅡ、MK2/cofilin信号通路有关,这可能是其发挥抗抑郁作用的重要机制。展开更多
目的探讨左归降糖解郁方调控小胶质细胞免疫受体分子样家族成员f(immune receptor molecule-like family member f,CD300f)改善糖尿病并发抑郁症(diabetes-related depression,DD)海马突触微环境损伤的保护作用及机制。方法原代分离、...目的探讨左归降糖解郁方调控小胶质细胞免疫受体分子样家族成员f(immune receptor molecule-like family member f,CD300f)改善糖尿病并发抑郁症(diabetes-related depression,DD)海马突触微环境损伤的保护作用及机制。方法原代分离、培养、纯化和鉴定小胶质细胞和海马神经元,复制模拟DD环境下的海马突触微环境损伤体外细胞模型,设置正常组、模型组、CD300f阻断剂CLM-1组、左归降糖解郁方含药血清组和左归降糖解郁方含药血清+CLM-1组。采用细胞成像分析观察小胶质细胞和海马神经元形态结构;采用ELISA法检测细胞上清液中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)、5-羟色胺(5-hydroxyteyptamine,5-HT)、多巴胺(dopamine,DA)含量;采用CCK-8实验及活死细胞染色评估海马神经元活性;采用尼氏染色观察海马神经元突触损伤;采用免疫荧光检测小胶质细胞中CD300f、Toll样受体4(Toll-like receptor 4,TLR4)和海马神经元中突触素(synaptophysin,SYN)、突触后密度蛋白95(postsynaptic density protein 95,PSD-95)蛋白表达;采用Western blotting检测SYN、PSD-95蛋白表达。结果左归降糖解郁方能有效改善小胶质细胞和海马神经元形态结构损伤,抑制细胞上清中神经炎症因子TNF-α、IL-1β和IDO水平(P<0.05、0.01),促进神经递质5-HT、DA水平(P<0.01),进而缓解海马神经元细胞活性及突触损伤。进一步的机制研究发现,左归降糖解郁方能显著增加海马小胶质细胞中CD300f表达(P<0.01),降低TLR4表达(P<0.05),并上调海马神经元中突触前膜SYN和突触后膜PSD-95表达(P<0.01),最终抑制海马突触微环境损伤。结论左归降糖解郁方能有效改善体外DD状态下的海马突触微环境损伤,其机制可能与上调小胶质细胞CD300f有关。展开更多
Objective To investigate the efficacy and mechanism of action of Compound Chaijin Jieyu Tablets(复方柴金解郁片,CCJJYT)in rats with insomnia complicated with depression.Methods Seventy-two Sprague-Dawley rats were rand...Objective To investigate the efficacy and mechanism of action of Compound Chaijin Jieyu Tablets(复方柴金解郁片,CCJJYT)in rats with insomnia complicated with depression.Methods Seventy-two Sprague-Dawley rats were randomly assigned into eight groups:the control,chronic unpredictable mild stress(CUMS),sleep deprivation(SD),CUMS+SD,positive drug(venlafaxine hydrochloride+diazepam),CCJJYT high-dose(CCJJYT˗2×),medium-dose(CCJJYT˗1×),and low-dose(CCJJYT˗0.5×)groups,with nine rats in each group.Depression-like behavior was evaluated by body weight,food intake,and behavioral tests such as the sucrose preference test(SPT),open field test(OFT),forced swimming test(FST),and pentobarbital-induced sleep test(PST).Hematoxylin-eosin(HE)staining and Golgi-Cox staining were used to observe changes in pathological tissue and synaptic morphology,respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of orexin-A and acetylcholine.The expression levels of orexin receptor 1(OXR1),melatonin receptor 1(MT1A),melatonin receptor 2(MT1B),acetylcholinesterase(AChE),and choline acetyltransferase(ChAT)were detected by immunohistochemistry and Western blot.Results In the present study,rats in the model group showed significant behavioral changes as well as a reduction in hippocampal dendritic branch length and synaptic number,along with increasing the content of orexin A and acetylcholine(P<0.05),and altered expression levels of OX1R,MT1A,MT1B,ChAT,and AChE in the hippocampus and prefrontal cortex after modeling(P<0.05).CCJJYT can improve depressive insomnia behavior and synaptic plasticity of rats(P<0.05),which is similar to that of the positive drug group.It can also decrease the content of orexin A and acetylcholine,and reduce the expression levels of OXR1 and ChAT in hippocampus and prefrontal cortex(P<0.05),and increase the expression levels of MT1A,MT1B,and AChE proteins(P<0.05).Conclusion CCJJYT has good antidepressant and insomnia effects,probably through the regulation of orexin-A,melatonin,and acetylcholine content in hippocampus and prefrontal cortex of rats,improving synaptic plasticity and thus exerting antidepressant and insomnia effects.展开更多
文摘盐诱导激酶(salt-inducible kinases,SIKs)属于腺苷酸活化激酶(AMP-activated protein kinase,AMPK)家族,主要调控环磷酸苷效应元件结合蛋白(cAMP-response element binding protein,CREB)转录共激活因子(CREB-regulated transcription coactivators,CRTCs)的胞核-胞质分布,对CRTC-CREB复合物的合成起到调节作用,从而间接影响CREB目的基因的转录与表达,影响多种生理过程,例如能量代谢、细胞周期进程和细胞凋亡。过去对SIKs的相关研究主要集中在外周系统疾病,包括肿瘤、高血压和糖异生等。近年来越来越多的研究开始探索SIKs和中枢神经系统疾病的关联性,如抑郁症、睡眠障碍、癫痫和阿尔茨海默病等都存在SIKs水平异常的情况,这提示SIKs信号失调参与这些疾病的病理过程,SIKs具有成为此类疾病新治疗靶点的潜力。本文就SIKs在抑郁症、睡眠障碍及其他神经系统疾病中的作用和调控机制进行综述。
文摘探讨柴金解郁安神片调控前扣带皮层(ACC)-腹侧海马(vHPC)谷氨酸能神经环路异常改善抑郁症大鼠腹侧海马神经元突触重塑的分子机制。首先运用化学遗传将谷氨酸能腺相关病毒(AAV)定位注射至大鼠ACC脑区,并通过慢性温和不可预知性应激(CUMS)联合孤笼饲养复制大鼠抑郁模型,实验设正常组、模型组、AAV空载组、AAV病毒组、AAV病毒+糖皮质激素受体(GR)阻断剂组、AAV病毒+趋化因子受体1(CX3CR1)阻断剂组、AAV病毒+柴金解郁安神片组,采用水迷宫(Morris water maze)、旷场(open-field)和强迫游泳(forced-swimming)实验联合动物行为分析系统评估大鼠抑郁样行为;苏木素-伊红(HE)染色检测大鼠ACC及vHPC脑区神经元形态结构变化;免疫荧光及核磷酸蛋白(c-Fos)检测大鼠ACC-vHPC谷氨酸能神经环路激活情况;高尔基染色和透射电镜检测大鼠vHPC神经元树突、树突棘及突触亚微结构变化;免疫荧光、Western blot分别检测大鼠vHPC谷氨酸能神经元细胞内突触重塑相关蛋白谷氨酸受体2A(GRIN2A)、谷氨酸受体2B(GRIN2B)、Ca^(2+)/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)、丝裂原激活蛋白激酶激活蛋白激酶2(MK2)、丝切蛋白(cofilin)表达水平。结果表明,谷氨酸能AAV病毒激活后模型组大鼠抑郁样行为表型、ACC及vHPC神经元形态结构、突触超微结构损伤更加加重,而GR、CX3CR1阻断剂均能不同程度逆转其异常改变,提示ACC脑区内胶质细胞GR/CX3CR1双信号介导的ACC-vHPC谷氨酸能神经环路异常激活可能与抑郁的发生发展密切相关。有趣的是,柴金解郁安神片也能显著抑制AAV病毒诱导的ACC-vHPC神经环路激活及Glu含量异常升高,同时有效逆转模型组大鼠进一步加重的抑郁样行为和vHPC谷氨酸能神经元突触重塑,并揭示其改善腹侧海马神经元突触损伤的分子机制可能与调控突触重塑相关信号NR/CaMKⅡ、MK2/cofilin有关。综上,该文证实了柴金解郁安神片能有效调控ACC-vHPC谷氨酸能神经环路异常进而改善抑郁症大鼠腹侧海马谷氨酸能神经元突触重塑,其分子机制可能与调节突触相关NR/CaMKⅡ、MK2/cofilin信号通路有关,这可能是其发挥抗抑郁作用的重要机制。
文摘目的探讨左归降糖解郁方调控小胶质细胞免疫受体分子样家族成员f(immune receptor molecule-like family member f,CD300f)改善糖尿病并发抑郁症(diabetes-related depression,DD)海马突触微环境损伤的保护作用及机制。方法原代分离、培养、纯化和鉴定小胶质细胞和海马神经元,复制模拟DD环境下的海马突触微环境损伤体外细胞模型,设置正常组、模型组、CD300f阻断剂CLM-1组、左归降糖解郁方含药血清组和左归降糖解郁方含药血清+CLM-1组。采用细胞成像分析观察小胶质细胞和海马神经元形态结构;采用ELISA法检测细胞上清液中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)、5-羟色胺(5-hydroxyteyptamine,5-HT)、多巴胺(dopamine,DA)含量;采用CCK-8实验及活死细胞染色评估海马神经元活性;采用尼氏染色观察海马神经元突触损伤;采用免疫荧光检测小胶质细胞中CD300f、Toll样受体4(Toll-like receptor 4,TLR4)和海马神经元中突触素(synaptophysin,SYN)、突触后密度蛋白95(postsynaptic density protein 95,PSD-95)蛋白表达;采用Western blotting检测SYN、PSD-95蛋白表达。结果左归降糖解郁方能有效改善小胶质细胞和海马神经元形态结构损伤,抑制细胞上清中神经炎症因子TNF-α、IL-1β和IDO水平(P<0.05、0.01),促进神经递质5-HT、DA水平(P<0.01),进而缓解海马神经元细胞活性及突触损伤。进一步的机制研究发现,左归降糖解郁方能显著增加海马小胶质细胞中CD300f表达(P<0.01),降低TLR4表达(P<0.05),并上调海马神经元中突触前膜SYN和突触后膜PSD-95表达(P<0.01),最终抑制海马突触微环境损伤。结论左归降糖解郁方能有效改善体外DD状态下的海马突触微环境损伤,其机制可能与上调小胶质细胞CD300f有关。
基金National Major New Drug Development Project(2017 ZX09309026)National Natural Science Foundation of China(82104846)+1 种基金Natural Science Foundation of Hunan Province(2022JJ40323 and 2022JJ80092)Natural Science Foundation of Changsha(Kq2202266).
文摘Objective To investigate the efficacy and mechanism of action of Compound Chaijin Jieyu Tablets(复方柴金解郁片,CCJJYT)in rats with insomnia complicated with depression.Methods Seventy-two Sprague-Dawley rats were randomly assigned into eight groups:the control,chronic unpredictable mild stress(CUMS),sleep deprivation(SD),CUMS+SD,positive drug(venlafaxine hydrochloride+diazepam),CCJJYT high-dose(CCJJYT˗2×),medium-dose(CCJJYT˗1×),and low-dose(CCJJYT˗0.5×)groups,with nine rats in each group.Depression-like behavior was evaluated by body weight,food intake,and behavioral tests such as the sucrose preference test(SPT),open field test(OFT),forced swimming test(FST),and pentobarbital-induced sleep test(PST).Hematoxylin-eosin(HE)staining and Golgi-Cox staining were used to observe changes in pathological tissue and synaptic morphology,respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of orexin-A and acetylcholine.The expression levels of orexin receptor 1(OXR1),melatonin receptor 1(MT1A),melatonin receptor 2(MT1B),acetylcholinesterase(AChE),and choline acetyltransferase(ChAT)were detected by immunohistochemistry and Western blot.Results In the present study,rats in the model group showed significant behavioral changes as well as a reduction in hippocampal dendritic branch length and synaptic number,along with increasing the content of orexin A and acetylcholine(P<0.05),and altered expression levels of OX1R,MT1A,MT1B,ChAT,and AChE in the hippocampus and prefrontal cortex after modeling(P<0.05).CCJJYT can improve depressive insomnia behavior and synaptic plasticity of rats(P<0.05),which is similar to that of the positive drug group.It can also decrease the content of orexin A and acetylcholine,and reduce the expression levels of OXR1 and ChAT in hippocampus and prefrontal cortex(P<0.05),and increase the expression levels of MT1A,MT1B,and AChE proteins(P<0.05).Conclusion CCJJYT has good antidepressant and insomnia effects,probably through the regulation of orexin-A,melatonin,and acetylcholine content in hippocampus and prefrontal cortex of rats,improving synaptic plasticity and thus exerting antidepressant and insomnia effects.
