背景:大麻素受体通过与配体结合,调控牙周炎的炎症和骨量,促进牙周组织的愈合,在临床上牙周炎的预防和治疗方面具有重要意义。目的:综述大麻素受体与牙周炎的关系,主要为大麻素Ⅰ型(CB1)受体、大麻素Ⅱ型(CB2)受体与炎症和牙槽骨骨改建...背景:大麻素受体通过与配体结合,调控牙周炎的炎症和骨量,促进牙周组织的愈合,在临床上牙周炎的预防和治疗方面具有重要意义。目的:综述大麻素受体与牙周炎的关系,主要为大麻素Ⅰ型(CB1)受体、大麻素Ⅱ型(CB2)受体与炎症和牙槽骨骨改建的关系,以及涉及的常见细胞信号传导通路,为牙周炎预防和治疗及其在临床其他领域的应用提供思路。方法:检索PubMed、万方数据库、CNKI中国期刊全文数据库1985年7月至2022年7月收录的相关文献。英文检索词为“cannabinoids receptor,CB1 receptor and periodontitis,CB2 receptor and periodontitis,CB1 receptors and bone remodeling,CB2 receptors and bone remodeling,CB1 receptors and signaling pathways,CB2 receptors and signaling pathways”,中文检索词为“大麻素受体,CB1受体和牙周炎,CB2受体和牙周炎,CB1受体和骨改建,CB2受体和骨改建,CB1受体和信号通路、CB2受体和信号通路”,最终纳入107篇文献进行归纳总结。结果与结论:①内源性大麻素系统包含多种受体,其中最具有代表性的为CB1和CB2受体,均属于G蛋白偶联超家族成员;二者在牙周组织中均存在表达;②在天然配体或人工合成激动剂的作用下,大麻素受体可通过不同的代谢通路在体内外产生特定的生理效应,从而调控牙周炎局部的炎症和骨细胞的生成和分化,最终影响炎症和骨量;③进一步研究大麻素受体与牙周炎炎症和牙槽骨骨形成、骨吸收的关系,以及涉及到的常见信号通路——丝裂原活化蛋白激酶(MAPK)信号通路、NF-κB信号通路,为临床上牙周炎的预防和治疗提供新的思路成为目前研究的重点。展开更多
Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports....Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports. In present study, we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system. This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide. In addition, the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95°C), forming a large molecular weight band when analyzed by immuno-blotting. Only denaturing temperatures ≤75°C yielded a clear band at the predicted molecular weight. Collectively, we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence, and described the requirement for a low sample denaturing temperature in immuno-blot analysis. These findings provide very useful information for efficient mammalian expression and immuno-blotting of membrane receptors.展开更多
The amygdala is an important hub for regulating emotions and is involved in the pathophysiology of many mental diseases,such as depression and anxiety.Meanwhile,the endocannabinoid system plays a crucial role in regul...The amygdala is an important hub for regulating emotions and is involved in the pathophysiology of many mental diseases,such as depression and anxiety.Meanwhile,the endocannabinoid system plays a crucial role in regulating emotions and mainly functions through the cannabinoid type-1 receptor(CB1R),which is strongly expressed in the amygdala of non-human primates(NHPs).However,it remains largely unknown how the CB1Rs in the amygdala of NHPs regulate mental diseases.Here,we investigated the role of CB1R by knocking down the cannabinoid receptor 1(CNR1)gene encoding CB1R in the amygdala of adult marmosets through regional delivery of AAV-SaCas9-gRNA.We found that CB1R knockdown in the amygdala induced anxiety-like behaviors,including disrupted night sleep,agitated psychomotor activity in new environments,and reduced social desire.Moreover,marmosets with CB1R-knockdown had up-regulated plasma cortisol levels.These results indicate that the knockdown of CB1Rs in the amygdala induces anxiety-like behaviors in marmosets,and this may be the mechanism underlying the regulation of anxiety by CB1Rs in the amygdala of NHPs.展开更多
Objective We aimed to investigate whether antagonism of the cannabinoid CB1 receptor(CB1R)could affect novel object recognition(NOR)memory in chronically rapid eye movement sleep-deprived(RSD)rats.Methods The animals ...Objective We aimed to investigate whether antagonism of the cannabinoid CB1 receptor(CB1R)could affect novel object recognition(NOR)memory in chronically rapid eye movement sleep-deprived(RSD)rats.Methods The animals were examined for recognition memory following a 7-day chronic partial RSD paradigm using the multiple platform technique.The CB1R antagonist rimonabant(1 or 3 mg/kg,i.p.)was administered either at one hour prior to the sample phase for acquisition,or immediately after the sample phase for consolidation,or at one hour before the test phase for retrieval of NOR memory.For the reconsolidation task,rimonabant was administered immediately after the second sample phase.Results The RSD episode impaired acquisition,consolidation,and retrieval,but it did not affect the reconsolidation of NOR memory.Rimonabant administration did not affect acquisition,consolidation,and reconsolidation;however,it attenuated impairment of the retrieval of NOR memory induced by chronic RSD.Conclusions These findings,along with our previous report,would seem to suggest that RSD may affect different phases of recognition memory based on its duration.Importantly,it seems that the CB1R may,at least in part,be involved in the adverse effects of chronic RSD on the retrieval,but not in the acquisition,consolidation,and reconsolidation,of NOR memory.展开更多
【目的】观察大麻素1型受体(CB1R)在C57B/L6小鼠神经元和胶质细胞的表达及其在实验性自身免疫性脑脊髓炎(EAE)小鼠中枢神经系统(CNS)和外周免疫器官的动态变化及其作用。【方法】取C57B/L6胎鼠的海马培养神经元及新生C57B/L6新生小鼠的...【目的】观察大麻素1型受体(CB1R)在C57B/L6小鼠神经元和胶质细胞的表达及其在实验性自身免疫性脑脊髓炎(EAE)小鼠中枢神经系统(CNS)和外周免疫器官的动态变化及其作用。【方法】取C57B/L6胎鼠的海马培养神经元及新生C57B/L6新生小鼠的皮层培养星形胶质细胞和小胶质细胞,观察CB1R蛋白的表达;观察正常对照、完全弗氏佐剂、EAE和特异性CB1R抑制剂(SR141716A,SR1)干预组小鼠的神经功能缺损症状和体质量变化;用real time PCR检测CB1R m RNA表达;Western blot和荧光免疫组织化学法检测CB1R蛋白表达;用ELISA法检测细胞因子IL-1β、IL-4、IL-10、IL-17、IL-6、TNF-α、IFN-γ浓度的变化。【结果】1C57B/L6小鼠神经元和小胶质细胞表达CB1R,星形胶质细胞不表达CB1R。2EAE小鼠神经元CB1R的表达显著低于CFA小鼠(P<0.01),小胶质细胞CB1R的表达显著高于CFA小鼠(P<0.01)。EAE小鼠脾脏中CB1R的表达与相同时点CFA小鼠无显著性差异(P>0.05)。3SR1干预促进EAE早发,上调EAE小鼠CNS和脾脏抗原特异性T细胞IL-17,TNF-α,IL-1β和IL-6的表达(P<0.05)。【结论】1神经元和小胶质细胞表达CB1R。2EAE小鼠脾脏中CB1R显著升高没有疾病特异性,可能与非特异性免疫激活有关。3中枢神经细胞上的CB1R可能兼有神经保护和免疫调节双重作用,参与EAE的发生和发展。展开更多
Cannabinoid CB1 receptors have been found in the superficial dorsal horn of the spinal cord, particularly the substantia gelatinosa (SG), which is thought to play a pivotal role in modulating nociceptive transmission....Cannabinoid CB1 receptors have been found in the superficial dorsal horn of the spinal cord, particularly the substantia gelatinosa (SG), which is thought to play a pivotal role in modulating nociceptive transmission. Although cannabinoids are known to inhibit excitatory transmission in SG neurons, their effects on inhibitory transmission have not yet been examined fully. In order to know further about a role of cannabinoids in regulating nociceptive transmission, we examined the effects of cannabinoids on inhibitory transmissions in adult rat SG neurons using whole-cell voltage-clamp recordings. Anandamide (10 μM) superfused for 2 min reduced glycinergic and GABAergic electrically-evoked inhibitory postsynaptic current (IPSC) amplitudes;these actions persisted for more than 6 min after washout. Similar actions were produced by cannabinoid-receptor agonist WIN55,212-2 (5 μM) and 2-arachidonoyl glycerol (20 μM). The evoked IPSC amplitudes reduced by anandamide recovered to the control level following superfusion of CB1-receptor antagonist SR141716A (5 μM). A ratio of the second to first evoked IPSC amplitude in paired-pulse experiments was increased by anandamide (10 μM). The frequencies of glycinergic and GABAergic spontaneous IPSCs were reduced by anandamide (10 μM) without a change in their amplitudes. It is concluded that cannabinoids depress inhibitory transmissions in adult rat SG neurons by activating CB1 receptors in nerve terminals. This action could contribute to the modulation of nociceptive transmission by cannabinoids.展开更多
Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and ...Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and surgery, among which the diet is still the most common way. However, none of these therapeutic measures available is ideal, making it necessary to find an effective medical treatment. The endocannabinoid system, which is well known for its contributions in certain mental processes such as relaxation, amelioration of pain and anxiety, and sedation initiation, has been recently reported to play an essential role in regulating appetite and metabolism to maintain energy balance, leading to the belief that endocannabinoid system is closely related to obesity. This new discovery deepens our understanding of obesity, and provides us with a new direction for clinical obesity treatment. Rimonabant is an antagonist for CB1, and has entered the market in some countries. However, although effective as an anti-obesity drug, rimonabant also causes obviously adverse side-effects, thus is being doubted and denied for medical usage.展开更多
OBJECTIVE To detect the underlying mechanism of time window for estrogen(E2)replacement treating cognitive decline.METHODS E2 begun 1 week after the ovariectomy(OVXST)or 3 months after the ovariectomy(OVXLT).Learning ...OBJECTIVE To detect the underlying mechanism of time window for estrogen(E2)replacement treating cognitive decline.METHODS E2 begun 1 week after the ovariectomy(OVXST)or 3 months after the ovariectomy(OVXLT).Learning and memory ability were examined by trace fear memory test and inhibitory avoidance test.LTP and LTD were detected by MED64.High throughput gene expression sequencing and microRNA(miR NA) sequencing were used to detecte the differently expressed genes between OVXSTand OVXLTafter estrogen treatment.RESULTS Subcutaneous injection of E2 improved fear memory formation in both 1 week after ovariectomy(OVXST) mice or 3 months after ovariectomy(OVXLT) mice.However,for fear memory extinction,facilitated by E2 in OVXSTmice,but impaired by E2 in OVXLTmice.Further researches showed in medial prefrontal cortex(mPFC),estrogen facilitates LTD in OVXSTmice but impairs LTD in OVXLTmice.Results of highthroughput sequencings of mR NA and miRNA in mPFC from sham,OVXSTmice,E2 treated OVXST mice,OVXLTmice,and E2 treated OVXLTmice indicated decreased miR-221-5 p expression in OVXLTmice compared with OVXSTmice.In OVXLT mice,miR-221-5 p could be further reduced by E2 treatment.Additionally,miR-221-5 p targeted neuralized E3 ubiquitin protein ligase 1 a/b(Neurl1 a/b) m RNA.Decreased miR-221-5 p will promotes cannabinoid receptor 1(CB1) ubiquitination through up-regulating Neurl1 a/b protein levels in E2 treated OVXLTmice,which disrupted the retrograde endocanabinoids system.Replenishing miR-221-5 p or treating with CB1 agonist rescued the fear extinction impairment in E2 treated OVXLTmice.CONCLUSION These results uncovered a epigenetic change after long term E2 responsible for failure of E2 improving cognitive performance in OVXLTmice,moreover miR-221-5 p and CB1 agonist as potential targets for prolonging the time window for E2 replacement therapy.展开更多
Background: Cannabinoid receptor subtype 1 (CB1) has a relationship to the proliferation of various cells including malignant tumoral cells. We investigated and compared the expression of CB1 in benign and malignant h...Background: Cannabinoid receptor subtype 1 (CB1) has a relationship to the proliferation of various cells including malignant tumoral cells. We investigated and compared the expression of CB1 in benign and malignant human prostate tissues and in benign and malignant human prostate cell lines, as well as its function for the proliferation of human prostate cancer cells. Methods: Real-time quantitative PCR was performed to compare its expressions in human prostate tissues (normal, benign hyperplasia, and cancer) and prostate cell lines (3 normal and 3 malignant). For localization of CB1, immunofluorescent staining with rabbit anti-CB1 polyclonal antibodies and tetramethyl isothiocyanate (TRITC)-labeled swine anti-rabbit immunoglobulin (DAKO) were used under fluorescence microscope. To further analyze whether cell death was induced by anandamide (non-selective agonist for CB1/CB2) via a receptor dependent mechanism, the viability of DU145 cells, which is known as androgen-insensitive prostate cancer cell, was measured using MTT assay. Results: CB1mRNA was found to be expressed in the all 3 human prostate tissues, however, CB1 protein was expressed in BPH and low grade malignant PC tissues, but not in high grade malignant PC tissues. CB1 as for cell lines, the expression of CB1 was low in malignant cell lines except for DU145. Anandamide elicited cell death, which was significantly inhibited by AM251 (selective antagonist for CB1), indicating that cell death induced by anandamide in DU145 cells was mediated by CB1. Anandamide time-dependently elicits up-regulation of CB1 in DU145 cells. Conclusions: CB1 may be an inhibitory regulator of androgen-insensitive human prostate cancer epithelial cell growth.展开更多
文摘背景:大麻素受体通过与配体结合,调控牙周炎的炎症和骨量,促进牙周组织的愈合,在临床上牙周炎的预防和治疗方面具有重要意义。目的:综述大麻素受体与牙周炎的关系,主要为大麻素Ⅰ型(CB1)受体、大麻素Ⅱ型(CB2)受体与炎症和牙槽骨骨改建的关系,以及涉及的常见细胞信号传导通路,为牙周炎预防和治疗及其在临床其他领域的应用提供思路。方法:检索PubMed、万方数据库、CNKI中国期刊全文数据库1985年7月至2022年7月收录的相关文献。英文检索词为“cannabinoids receptor,CB1 receptor and periodontitis,CB2 receptor and periodontitis,CB1 receptors and bone remodeling,CB2 receptors and bone remodeling,CB1 receptors and signaling pathways,CB2 receptors and signaling pathways”,中文检索词为“大麻素受体,CB1受体和牙周炎,CB2受体和牙周炎,CB1受体和骨改建,CB2受体和骨改建,CB1受体和信号通路、CB2受体和信号通路”,最终纳入107篇文献进行归纳总结。结果与结论:①内源性大麻素系统包含多种受体,其中最具有代表性的为CB1和CB2受体,均属于G蛋白偶联超家族成员;二者在牙周组织中均存在表达;②在天然配体或人工合成激动剂的作用下,大麻素受体可通过不同的代谢通路在体内外产生特定的生理效应,从而调控牙周炎局部的炎症和骨细胞的生成和分化,最终影响炎症和骨量;③进一步研究大麻素受体与牙周炎炎症和牙槽骨骨形成、骨吸收的关系,以及涉及到的常见信号通路——丝裂原活化蛋白激酶(MAPK)信号通路、NF-κB信号通路,为临床上牙周炎的预防和治疗提供新的思路成为目前研究的重点。
基金supported by a grant from Army Medical Research Program of China(No.08G168)
文摘Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports. In present study, we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system. This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide. In addition, the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95°C), forming a large molecular weight band when analyzed by immuno-blotting. Only denaturing temperatures ≤75°C yielded a clear band at the predicted molecular weight. Collectively, we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence, and described the requirement for a low sample denaturing temperature in immuno-blot analysis. These findings provide very useful information for efficient mammalian expression and immuno-blotting of membrane receptors.
基金supported by the Zhejiang Province Natural Science Foundation of China(LD22H090003)Key-Area Research and Development Program of Guangdong Province(2019B030335001 and 2018B030334001)+3 种基金the National Natural Science Foundation of China(31871070,82090031,32071097,31871056,and 32170991)the Key R&D Program of Zhejiang Province(2020C03009)Fundamental Research Funds for the Central Universities(2021FZZX001-37)the CAMS Innovation Fund for Medical Sciences(2019-I2M-5-057).
文摘The amygdala is an important hub for regulating emotions and is involved in the pathophysiology of many mental diseases,such as depression and anxiety.Meanwhile,the endocannabinoid system plays a crucial role in regulating emotions and mainly functions through the cannabinoid type-1 receptor(CB1R),which is strongly expressed in the amygdala of non-human primates(NHPs).However,it remains largely unknown how the CB1Rs in the amygdala of NHPs regulate mental diseases.Here,we investigated the role of CB1R by knocking down the cannabinoid receptor 1(CNR1)gene encoding CB1R in the amygdala of adult marmosets through regional delivery of AAV-SaCas9-gRNA.We found that CB1R knockdown in the amygdala induced anxiety-like behaviors,including disrupted night sleep,agitated psychomotor activity in new environments,and reduced social desire.Moreover,marmosets with CB1R-knockdown had up-regulated plasma cortisol levels.These results indicate that the knockdown of CB1Rs in the amygdala induces anxiety-like behaviors in marmosets,and this may be the mechanism underlying the regulation of anxiety by CB1Rs in the amygdala of NHPs.
基金Supported by the Research Council of Kermanshah University of Medical Sciences,Kermanshah,Iran for financial support(grant no.:990812).
文摘Objective We aimed to investigate whether antagonism of the cannabinoid CB1 receptor(CB1R)could affect novel object recognition(NOR)memory in chronically rapid eye movement sleep-deprived(RSD)rats.Methods The animals were examined for recognition memory following a 7-day chronic partial RSD paradigm using the multiple platform technique.The CB1R antagonist rimonabant(1 or 3 mg/kg,i.p.)was administered either at one hour prior to the sample phase for acquisition,or immediately after the sample phase for consolidation,or at one hour before the test phase for retrieval of NOR memory.For the reconsolidation task,rimonabant was administered immediately after the second sample phase.Results The RSD episode impaired acquisition,consolidation,and retrieval,but it did not affect the reconsolidation of NOR memory.Rimonabant administration did not affect acquisition,consolidation,and reconsolidation;however,it attenuated impairment of the retrieval of NOR memory induced by chronic RSD.Conclusions These findings,along with our previous report,would seem to suggest that RSD may affect different phases of recognition memory based on its duration.Importantly,it seems that the CB1R may,at least in part,be involved in the adverse effects of chronic RSD on the retrieval,but not in the acquisition,consolidation,and reconsolidation,of NOR memory.
文摘【目的】观察大麻素1型受体(CB1R)在C57B/L6小鼠神经元和胶质细胞的表达及其在实验性自身免疫性脑脊髓炎(EAE)小鼠中枢神经系统(CNS)和外周免疫器官的动态变化及其作用。【方法】取C57B/L6胎鼠的海马培养神经元及新生C57B/L6新生小鼠的皮层培养星形胶质细胞和小胶质细胞,观察CB1R蛋白的表达;观察正常对照、完全弗氏佐剂、EAE和特异性CB1R抑制剂(SR141716A,SR1)干预组小鼠的神经功能缺损症状和体质量变化;用real time PCR检测CB1R m RNA表达;Western blot和荧光免疫组织化学法检测CB1R蛋白表达;用ELISA法检测细胞因子IL-1β、IL-4、IL-10、IL-17、IL-6、TNF-α、IFN-γ浓度的变化。【结果】1C57B/L6小鼠神经元和小胶质细胞表达CB1R,星形胶质细胞不表达CB1R。2EAE小鼠神经元CB1R的表达显著低于CFA小鼠(P<0.01),小胶质细胞CB1R的表达显著高于CFA小鼠(P<0.01)。EAE小鼠脾脏中CB1R的表达与相同时点CFA小鼠无显著性差异(P>0.05)。3SR1干预促进EAE早发,上调EAE小鼠CNS和脾脏抗原特异性T细胞IL-17,TNF-α,IL-1β和IL-6的表达(P<0.05)。【结论】1神经元和小胶质细胞表达CB1R。2EAE小鼠脾脏中CB1R显著升高没有疾病特异性,可能与非特异性免疫激活有关。3中枢神经细胞上的CB1R可能兼有神经保护和免疫调节双重作用,参与EAE的发生和发展。
文摘Cannabinoid CB1 receptors have been found in the superficial dorsal horn of the spinal cord, particularly the substantia gelatinosa (SG), which is thought to play a pivotal role in modulating nociceptive transmission. Although cannabinoids are known to inhibit excitatory transmission in SG neurons, their effects on inhibitory transmission have not yet been examined fully. In order to know further about a role of cannabinoids in regulating nociceptive transmission, we examined the effects of cannabinoids on inhibitory transmissions in adult rat SG neurons using whole-cell voltage-clamp recordings. Anandamide (10 μM) superfused for 2 min reduced glycinergic and GABAergic electrically-evoked inhibitory postsynaptic current (IPSC) amplitudes;these actions persisted for more than 6 min after washout. Similar actions were produced by cannabinoid-receptor agonist WIN55,212-2 (5 μM) and 2-arachidonoyl glycerol (20 μM). The evoked IPSC amplitudes reduced by anandamide recovered to the control level following superfusion of CB1-receptor antagonist SR141716A (5 μM). A ratio of the second to first evoked IPSC amplitude in paired-pulse experiments was increased by anandamide (10 μM). The frequencies of glycinergic and GABAergic spontaneous IPSCs were reduced by anandamide (10 μM) without a change in their amplitudes. It is concluded that cannabinoids depress inhibitory transmissions in adult rat SG neurons by activating CB1 receptors in nerve terminals. This action could contribute to the modulation of nociceptive transmission by cannabinoids.
基金supported by the Study Grant (2007) for Medical Students of Second Military Medical University,China
文摘Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and surgery, among which the diet is still the most common way. However, none of these therapeutic measures available is ideal, making it necessary to find an effective medical treatment. The endocannabinoid system, which is well known for its contributions in certain mental processes such as relaxation, amelioration of pain and anxiety, and sedation initiation, has been recently reported to play an essential role in regulating appetite and metabolism to maintain energy balance, leading to the belief that endocannabinoid system is closely related to obesity. This new discovery deepens our understanding of obesity, and provides us with a new direction for clinical obesity treatment. Rimonabant is an antagonist for CB1, and has entered the market in some countries. However, although effective as an anti-obesity drug, rimonabant also causes obviously adverse side-effects, thus is being doubted and denied for medical usage.
文摘OBJECTIVE To detect the underlying mechanism of time window for estrogen(E2)replacement treating cognitive decline.METHODS E2 begun 1 week after the ovariectomy(OVXST)or 3 months after the ovariectomy(OVXLT).Learning and memory ability were examined by trace fear memory test and inhibitory avoidance test.LTP and LTD were detected by MED64.High throughput gene expression sequencing and microRNA(miR NA) sequencing were used to detecte the differently expressed genes between OVXSTand OVXLTafter estrogen treatment.RESULTS Subcutaneous injection of E2 improved fear memory formation in both 1 week after ovariectomy(OVXST) mice or 3 months after ovariectomy(OVXLT) mice.However,for fear memory extinction,facilitated by E2 in OVXSTmice,but impaired by E2 in OVXLTmice.Further researches showed in medial prefrontal cortex(mPFC),estrogen facilitates LTD in OVXSTmice but impairs LTD in OVXLTmice.Results of highthroughput sequencings of mR NA and miRNA in mPFC from sham,OVXSTmice,E2 treated OVXST mice,OVXLTmice,and E2 treated OVXLTmice indicated decreased miR-221-5 p expression in OVXLTmice compared with OVXSTmice.In OVXLT mice,miR-221-5 p could be further reduced by E2 treatment.Additionally,miR-221-5 p targeted neuralized E3 ubiquitin protein ligase 1 a/b(Neurl1 a/b) m RNA.Decreased miR-221-5 p will promotes cannabinoid receptor 1(CB1) ubiquitination through up-regulating Neurl1 a/b protein levels in E2 treated OVXLTmice,which disrupted the retrograde endocanabinoids system.Replenishing miR-221-5 p or treating with CB1 agonist rescued the fear extinction impairment in E2 treated OVXLTmice.CONCLUSION These results uncovered a epigenetic change after long term E2 responsible for failure of E2 improving cognitive performance in OVXLTmice,moreover miR-221-5 p and CB1 agonist as potential targets for prolonging the time window for E2 replacement therapy.
文摘Background: Cannabinoid receptor subtype 1 (CB1) has a relationship to the proliferation of various cells including malignant tumoral cells. We investigated and compared the expression of CB1 in benign and malignant human prostate tissues and in benign and malignant human prostate cell lines, as well as its function for the proliferation of human prostate cancer cells. Methods: Real-time quantitative PCR was performed to compare its expressions in human prostate tissues (normal, benign hyperplasia, and cancer) and prostate cell lines (3 normal and 3 malignant). For localization of CB1, immunofluorescent staining with rabbit anti-CB1 polyclonal antibodies and tetramethyl isothiocyanate (TRITC)-labeled swine anti-rabbit immunoglobulin (DAKO) were used under fluorescence microscope. To further analyze whether cell death was induced by anandamide (non-selective agonist for CB1/CB2) via a receptor dependent mechanism, the viability of DU145 cells, which is known as androgen-insensitive prostate cancer cell, was measured using MTT assay. Results: CB1mRNA was found to be expressed in the all 3 human prostate tissues, however, CB1 protein was expressed in BPH and low grade malignant PC tissues, but not in high grade malignant PC tissues. CB1 as for cell lines, the expression of CB1 was low in malignant cell lines except for DU145. Anandamide elicited cell death, which was significantly inhibited by AM251 (selective antagonist for CB1), indicating that cell death induced by anandamide in DU145 cells was mediated by CB1. Anandamide time-dependently elicits up-regulation of CB1 in DU145 cells. Conclusions: CB1 may be an inhibitory regulator of androgen-insensitive human prostate cancer epithelial cell growth.
