Direct in vivo conversion of astrocytes into functional new neurons induced by neural transcription factors has been recognized as a potential new therapeutic intervention for neural injury and degenerative disorders....Direct in vivo conversion of astrocytes into functional new neurons induced by neural transcription factors has been recognized as a potential new therapeutic intervention for neural injury and degenerative disorders. However, a few recent studies have claimed that neural transcription factors cannot convert astrocytes into neurons, attributing the converted neurons to pre-existing neurons mis-expressing transgenes. In this study, we overexpressed three distinct neural transcription factors––NeuroD1, Ascl1, and Dlx2––in reactive astrocytes in mouse cortices subjected to stab injury, resulting in a series of significant changes in astrocyte properties. Initially, the three neural transcription factors were exclusively expressed in the nuclei of astrocytes. Over time, however, these astrocytes gradually adopted neuronal morphology, and the neural transcription factors was gradually observed in the nuclei of neuron-like cells instead of astrocytes. Furthermore,we noted that transcription factor-infected astrocytes showed a progressive decrease in the expression of astrocytic markers AQP4(astrocyte endfeet signal), CX43(gap junction signal), and S100β. Importantly, none of these changes could be attributed to transgene leakage into preexisting neurons. Therefore, our findings suggest that neural transcription factors such as NeuroD1, Ascl1, and Dlx2 can effectively convert reactive astrocytes into neurons in the adult mammalian brain.展开更多
The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were stu...The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.展开更多
目的:探讨通窍胶囊联合布地奈德对分泌性中耳炎(SOM)患者听力及血清水通道蛋白(AQP)-1、AQP-4水平的影响。方法:选取2015年4月至2016年8月延安大学咸阳医院收治的SOM患者114例,按照随机数字表法分为对照组(n=56)和观察组(n=58),对照组...目的:探讨通窍胶囊联合布地奈德对分泌性中耳炎(SOM)患者听力及血清水通道蛋白(AQP)-1、AQP-4水平的影响。方法:选取2015年4月至2016年8月延安大学咸阳医院收治的SOM患者114例,按照随机数字表法分为对照组(n=56)和观察组(n=58),对照组患者给予常规治疗和布地奈德鼓室注射治疗,观察组在对照组基础上给予通窍胶囊治疗,观察并比较两组患者治疗前、后在0.5 k Hz、1 k Hz、2 k Hz频率下的气导阈值及血清AQP-1、AQP-4水平的变化。结果:治疗前,两组患者各个频率下的气导阈值比较差异均无统计学意义(均P>0.05);观察组治疗后在0.5 k Hz、1 k Hz、2 k Hz频率下的气导阈值均明显低于对照组(均P<0.01)。观察组治疗后血清AQP-1水平为(11.02±1.04)μg/L,明显高于对照组的(7.26±0.34)μg/L(P<0.01),而两组血清AQP-4水平比较差异无统计学意义(P>0.05)。结论:通窍胶囊联合布地奈德治疗SOM,可有效改善患者听力,提高患者血清AQP-1水平,改善耳道分泌情况。展开更多
AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparen...AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.展开更多
基金supported by the Key Project of Guangzhou City,No.202206060002Science and Technology Project of Guangdong Province,No.2018B030332001Guangdong Provincial Pearl River Project,No.2021ZT09Y552 (all to GC)。
文摘Direct in vivo conversion of astrocytes into functional new neurons induced by neural transcription factors has been recognized as a potential new therapeutic intervention for neural injury and degenerative disorders. However, a few recent studies have claimed that neural transcription factors cannot convert astrocytes into neurons, attributing the converted neurons to pre-existing neurons mis-expressing transgenes. In this study, we overexpressed three distinct neural transcription factors––NeuroD1, Ascl1, and Dlx2––in reactive astrocytes in mouse cortices subjected to stab injury, resulting in a series of significant changes in astrocyte properties. Initially, the three neural transcription factors were exclusively expressed in the nuclei of astrocytes. Over time, however, these astrocytes gradually adopted neuronal morphology, and the neural transcription factors was gradually observed in the nuclei of neuron-like cells instead of astrocytes. Furthermore,we noted that transcription factor-infected astrocytes showed a progressive decrease in the expression of astrocytic markers AQP4(astrocyte endfeet signal), CX43(gap junction signal), and S100β. Importantly, none of these changes could be attributed to transgene leakage into preexisting neurons. Therefore, our findings suggest that neural transcription factors such as NeuroD1, Ascl1, and Dlx2 can effectively convert reactive astrocytes into neurons in the adult mammalian brain.
文摘The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.
文摘目的:探讨通窍胶囊联合布地奈德对分泌性中耳炎(SOM)患者听力及血清水通道蛋白(AQP)-1、AQP-4水平的影响。方法:选取2015年4月至2016年8月延安大学咸阳医院收治的SOM患者114例,按照随机数字表法分为对照组(n=56)和观察组(n=58),对照组患者给予常规治疗和布地奈德鼓室注射治疗,观察组在对照组基础上给予通窍胶囊治疗,观察并比较两组患者治疗前、后在0.5 k Hz、1 k Hz、2 k Hz频率下的气导阈值及血清AQP-1、AQP-4水平的变化。结果:治疗前,两组患者各个频率下的气导阈值比较差异均无统计学意义(均P>0.05);观察组治疗后在0.5 k Hz、1 k Hz、2 k Hz频率下的气导阈值均明显低于对照组(均P<0.01)。观察组治疗后血清AQP-1水平为(11.02±1.04)μg/L,明显高于对照组的(7.26±0.34)μg/L(P<0.01),而两组血清AQP-4水平比较差异无统计学意义(P>0.05)。结论:通窍胶囊联合布地奈德治疗SOM,可有效改善患者听力,提高患者血清AQP-1水平,改善耳道分泌情况。
基金Supported by the National Natural Science Foundation of China(No.81070715)Innovative Platform Foundation of Fujian ProvinceChina(No.2010Y2003)
文摘AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.
