目的:探讨丹龙醒脑方对局灶性脑缺血再灌注大鼠侧脑室室管膜下区(SVZ)神经干细胞(NSCs)增殖与Frizzled3、Dsh1、β-catenin表达的关系。方法:将120只雄性SD大鼠随机分为假手术组、模型组、依达组、丹龙组。后3组用线栓法制备局灶性脑缺...目的:探讨丹龙醒脑方对局灶性脑缺血再灌注大鼠侧脑室室管膜下区(SVZ)神经干细胞(NSCs)增殖与Frizzled3、Dsh1、β-catenin表达的关系。方法:将120只雄性SD大鼠随机分为假手术组、模型组、依达组、丹龙组。后3组用线栓法制备局灶性脑缺血再灌注(MCAO/R)模型,再灌注7d后取缺血侧SVZ脑组织。采用Brdu免疫荧光法检测SVZ区NSCs增殖,RT-q PCR法、Western Blot法分别检测Frizzled3、Dsh1、β-catenin m RNA和蛋白的表达。结果:与假手术组比较,其余各组Brdu阳性细胞数明显增多,Frizzled3、Dsh1、β-catenin蛋白及m RNA的表达明显升高(P<0.05,P<0.01);与模型组比较,依达组、丹龙组Brdu阳性细胞数明显增多,Dsh1、β-catenin m RNA及蛋白的表达明显增强(P<0.05,P<0.01),丹龙组Frizzled3表达明显增强(P<0.05,P<0.01),但依达组不明显。依达组、丹龙组组间比较,除Frizzled3蛋白表达丹龙组要优于依达组(P<0.01),其余指标均无明显差异。结论:丹龙醒脑方能促进脑缺血后SVZ区NSCs增殖,其机制可能与通过上调Frizzled3和Dsh1的水平,进而促进Wnt信号的传递和增强关键因子β-catenin的表达有关。展开更多
Uncaria rhynchophylla is commonly recognized as a traditional treatment for dizziness,cerebrovascular diseases,and nervous disorders in China.Previously,the neuro-protective activities of the alkaloids from U.rhyncho...Uncaria rhynchophylla is commonly recognized as a traditional treatment for dizziness,cerebrovascular diseases,and nervous disorders in China.Previously,the neuro-protective activities of the alkaloids from U.rhynchophylla were intensively reported.In current work,three new indole alkaloids(1–3),identified as geissoschizic acid(1),geissoschizic acid N4-oxide(2),and 3b-sitsirikine N4-oxide(3),as well as 26 known analogues were isolated from U.rhynchophylla.However,in the neural stem cells(NSCs)proliferation assay for all isolated compounds,geissoschizic acid(1),geissoschizic acid N4-oxide(2),isocorynoxeine(6),isorhynchophylline(7),(4S)-akuammigine N-oxide(8),and(4S)-rhynchophylline N-oxide(10)showed unexpected inhibitory activities at 10 μM.Unlike previous neuro-protective reports,as a warning or caution,our finding showcased a clue for possible NSCs toxicity and the neural lesions risk of U.rhynchophylla,while the structure–activity relationships of the isolated compounds were discussed also.展开更多
The purpose of the study was to investigate the impact of rat cytomegalovirus （RCMV） infection on the development of the nervous system in rat embryos, and to evaluate the involvement of Wnt signaling pathway key mo...The purpose of the study was to investigate the impact of rat cytomegalovirus （RCMV） infection on the development of the nervous system in rat embryos, and to evaluate the involvement of Wnt signaling pathway key molecules and the downstream gene neurogenin 1 （Ngnl） in RCMV infected neural stem cells （NSCs）. Infection and control groups were established, each containing 20 pregnant Wistar rats. Rats in the infection group were inoculated with RCMV by intraperitoneal injection on the first day of pregnancy. Rat E20 embryos were taken to evaluate the teratogenic rate. NSCs were isolated from El3 embryos, and maintained in vitro. We found： 1） Poor fetal development was found in the infection group with low survival and high malformation rates. 2） The proliferation and differentiation of NSCs were affected. In the infection group, NSCs proliferated more slowly and had a lower neurosphere formation rate than the control. The differentiation ratio from NSCs to neurons and glial cells was significantly different from that of the control, showed by immunofluorescenee staining. 3） Ngnl mRNA expression and the nuclear p-catenin protein level were significantly lower than the control on day 2 when NSCs differentiated. 4） The Morris water maze test was performed on 4-week pups, and the infected rats were found worse in learning and memory ability. In a summary, RCMV infection caused abnormalities in the rat embryonic nervous system, significantly inhibited NSC proliferation and differentiation, and inhibited the expression of key molecules in the Wnt/β-catenin signaling pathway so as to affect NSCs differentiation. This may be an important mechanism by which RCMV causes embryonic nervous system abnormalities.展开更多
The regeneration of the injured nerve and recovery of its function have brought attention in the medical field. Electrical stimulation(ES) can enhance the cellular biological behavior and has been widely studied in th...The regeneration of the injured nerve and recovery of its function have brought attention in the medical field. Electrical stimulation(ES) can enhance the cellular biological behavior and has been widely studied in the treatment of neurological diseases. Microfluidic technology can provide a cell culture platform with the well-controlled environment. Here a novel microfluidic/microelectrode composite microdevice was developed by embedding the microelectrodes to the microfluidic platform, in which microfluidics provided a controlled cell culture platform, and ES promoted the NSCs proliferation. We performed ES on rat neural stem cells(NSCs) to observe the effect on their growth, differentiation, proliferation, and preliminary explored the ES influence on cells in vitro. The results of immunofluorescence showed that ES had no significant effect on the NSCs specific expression, and the NSCs specific expression reached 98.9%± 0.4% after three days of ES. In addition, ES significantly promoted cell growth and the cell proliferation rate reached 49.41%. To conclude, the microfluidic/microelectrode composite microdevice can play a positive role in the nerve injury repair and fundamental research of neurological diseases.展开更多
文摘目的:探讨丹龙醒脑方对局灶性脑缺血再灌注大鼠侧脑室室管膜下区(SVZ)神经干细胞(NSCs)增殖与Frizzled3、Dsh1、β-catenin表达的关系。方法:将120只雄性SD大鼠随机分为假手术组、模型组、依达组、丹龙组。后3组用线栓法制备局灶性脑缺血再灌注(MCAO/R)模型,再灌注7d后取缺血侧SVZ脑组织。采用Brdu免疫荧光法检测SVZ区NSCs增殖,RT-q PCR法、Western Blot法分别检测Frizzled3、Dsh1、β-catenin m RNA和蛋白的表达。结果:与假手术组比较,其余各组Brdu阳性细胞数明显增多,Frizzled3、Dsh1、β-catenin蛋白及m RNA的表达明显升高(P<0.05,P<0.01);与模型组比较,依达组、丹龙组Brdu阳性细胞数明显增多,Dsh1、β-catenin m RNA及蛋白的表达明显增强(P<0.05,P<0.01),丹龙组Frizzled3表达明显增强(P<0.05,P<0.01),但依达组不明显。依达组、丹龙组组间比较,除Frizzled3蛋白表达丹龙组要优于依达组(P<0.01),其余指标均无明显差异。结论:丹龙醒脑方能促进脑缺血后SVZ区NSCs增殖,其机制可能与通过上调Frizzled3和Dsh1的水平,进而促进Wnt信号的传递和增强关键因子β-catenin的表达有关。
基金The authors are grateful to the National Natural Science Foundation of China(81225024,31500292,31770388)for financial support.
文摘Uncaria rhynchophylla is commonly recognized as a traditional treatment for dizziness,cerebrovascular diseases,and nervous disorders in China.Previously,the neuro-protective activities of the alkaloids from U.rhynchophylla were intensively reported.In current work,three new indole alkaloids(1–3),identified as geissoschizic acid(1),geissoschizic acid N4-oxide(2),and 3b-sitsirikine N4-oxide(3),as well as 26 known analogues were isolated from U.rhynchophylla.However,in the neural stem cells(NSCs)proliferation assay for all isolated compounds,geissoschizic acid(1),geissoschizic acid N4-oxide(2),isocorynoxeine(6),isorhynchophylline(7),(4S)-akuammigine N-oxide(8),and(4S)-rhynchophylline N-oxide(10)showed unexpected inhibitory activities at 10 μM.Unlike previous neuro-protective reports,as a warning or caution,our finding showcased a clue for possible NSCs toxicity and the neural lesions risk of U.rhynchophylla,while the structure–activity relationships of the isolated compounds were discussed also.
基金Shandong Province High-level Talent of Health 1020 Project Fund(No.2008-1)Science and Technology Creative Research of Weifang Medical University(No.K11TS1010)+1 种基金A Project of Shandong Province Higher Educational Science and Technology Program(No.J12LK04)National Natural Science Foundation of China(30900775)
文摘The purpose of the study was to investigate the impact of rat cytomegalovirus （RCMV） infection on the development of the nervous system in rat embryos, and to evaluate the involvement of Wnt signaling pathway key molecules and the downstream gene neurogenin 1 （Ngnl） in RCMV infected neural stem cells （NSCs）. Infection and control groups were established, each containing 20 pregnant Wistar rats. Rats in the infection group were inoculated with RCMV by intraperitoneal injection on the first day of pregnancy. Rat E20 embryos were taken to evaluate the teratogenic rate. NSCs were isolated from El3 embryos, and maintained in vitro. We found： 1） Poor fetal development was found in the infection group with low survival and high malformation rates. 2） The proliferation and differentiation of NSCs were affected. In the infection group, NSCs proliferated more slowly and had a lower neurosphere formation rate than the control. The differentiation ratio from NSCs to neurons and glial cells was significantly different from that of the control, showed by immunofluorescenee staining. 3） Ngnl mRNA expression and the nuclear p-catenin protein level were significantly lower than the control on day 2 when NSCs differentiated. 4） The Morris water maze test was performed on 4-week pups, and the infected rats were found worse in learning and memory ability. In a summary, RCMV infection caused abnormalities in the rat embryonic nervous system, significantly inhibited NSC proliferation and differentiation, and inhibited the expression of key molecules in the Wnt/β-catenin signaling pathway so as to affect NSCs differentiation. This may be an important mechanism by which RCMV causes embryonic nervous system abnormalities.
基金financially supported by the Key Scientific and Technological Projects of the Beijing Education Commission (No.KZ201910005009)。
文摘The regeneration of the injured nerve and recovery of its function have brought attention in the medical field. Electrical stimulation(ES) can enhance the cellular biological behavior and has been widely studied in the treatment of neurological diseases. Microfluidic technology can provide a cell culture platform with the well-controlled environment. Here a novel microfluidic/microelectrode composite microdevice was developed by embedding the microelectrodes to the microfluidic platform, in which microfluidics provided a controlled cell culture platform, and ES promoted the NSCs proliferation. We performed ES on rat neural stem cells(NSCs) to observe the effect on their growth, differentiation, proliferation, and preliminary explored the ES influence on cells in vitro. The results of immunofluorescence showed that ES had no significant effect on the NSCs specific expression, and the NSCs specific expression reached 98.9%± 0.4% after three days of ES. In addition, ES significantly promoted cell growth and the cell proliferation rate reached 49.41%. To conclude, the microfluidic/microelectrode composite microdevice can play a positive role in the nerve injury repair and fundamental research of neurological diseases.
