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Effects of huperzine A on secretion of nerve growth factor in cultured rat cortical astrocytes and neurite outgrowth in rat PC12 cells

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Acta Pharmacologica Sinica 2005年第6期26卷 673-678页

作者： Li-li TANG, Rui WANG, Xi-can TANG~2 State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Aim: To study the effects of huperzine A (HupA) on neuritogenic activity and the expression of nerve growth factor (NGF). Methods: After being treated with 10 μmol/L HupA, neurite outgrowth of PC 12 cells was observed and counted under phase-contrast microscopy. Mitogenic activity was assayed by [~3H]thymidine incorporation. Cell cytotoxicity was evaluated by lactate dehydrogenase (LDH) release. AChE activity, mRNA and protein expression were measured by the Ellman’s method, RT-PCR, and Western blot, respectively. NGF mRNA and protein levels were determined by RT-PCR and ELISA assays. Results: Treatment of PC 12 cells with 10 渭mol/L HupA for 48 h markedly increased the number of neurite-bearing cells, but caused no significant alteration in cell viability or other signs of cytotoxicity. In addition to inhibiting AChE activity, 10 渭mol/L HupA also increased the mRNA and protein levels of this enzyme. In addition, following 2 h exposure of the astrocytes to 10 渭mol/L HupA, there was a significant up-regula-tion of mRNA for NGF and P75 low-affinity NGF receptor. The protein level of NGF was also increased after 24 h treatment with HupA. Conclusion: Our findings demonstrate for the first time that HupA has a direct or indirect neurotrophic activity, which might be beneficial in treatment of neurodegenerative disorders such as Alzheimer disease.

关键词： huperzine A nerve growth factor neurite outgrowth: acetylcholinesterase inhibitors

Intranasal nerve growth factor bypasses the blood-brain barrier and affects spinal cord neurons in spinal cord injury

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Neural Regeneration Research 2014年第10期9卷 1025-1030页

作者： Luigi Aloe Patrizia Bianchi Alberto De Bellis Marzia Soligo Maria Luisa Rocco Cellular Biology and Neurobiology Institute National Research Council(CNR) Maria Rosaria Maglione Foundation Onlus NapoliItaly Intitute of Translational Pharmacology National Research Council(CNR)

The purpose of this work was to investigate whether, by intranasal administration, the nerve growth factor bypasses the blood-brain barrier and turns over the spinal cord neurons and if such therapeutic approach could be of value in the treatment of spinal cord injury. Adult Sprague-Dawley rats with intact and injured spinal cord received daily intranasal nerve growth factor administration in both nostrils for 1 day or for 3 consecutive weeks. We found an in-creased content of nerve growth factor and enhanced expression of nerve growth factor receptor in the spinal cord 24 hours after a single intranasal administration of nerve growth factor in healthy rats, while daily treatment for 3 weeks in a model of spinal cord injury improved the deifcits in locomotor behaviour and increased spinal content of both nerve growth factor and nerve growth factor receptors. These outcomes suggest that the intranasal nerve growth factor bypasses blood-brain barrier and affects spinal cord neurons in spinal cord injury. They also suggest exploiting the possible therapeutic role of intranasally delivered nerve growth factor for the neuroprotection of damaged spinal nerve cells.

关键词： nerve regeneration spinal cord injury nerve growth factor intranasal delivery blood-brain barrier motor function leptin neuroprotection rats neural regeneration

Therapeutic effect of nerve growth factor on cerebral infarction in dogs using the hemisphere anomalous volume ratio of diffusion-weighted magnetic resonance imaging

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Neural Regeneration Research 2012年第24期7卷 1873-1880页

作者： Yong Wang Hui Zhang Zhe Wang Zuojun Geng Huaijun Liu Haiqing Yang Peng Song Qing Liu Department of Radiology the Second Hospital of Hebei Medical UniversityShijiazhuang 050000Hebei ProvinceChina Department of Radiology Hebei General HospitalShijiazhuang 050051Hebei ProvinceChina Department of Computer Science Institute of Basic MedicineHebei Medical UniversityShijiazhuang 050031Hebei ProvinceChina

A model of focal cerebral ischemic infarction was established in dogs through middle cerebral artery occlusion of the right *** minutes after occlusion,models were injected with nerve growth factor adjacent to the infarct *** therapeutic effect of nerve growth factor against cerebral infarction was assessed using the hemisphere anomalous volume ratio,a quantitative index of diffusion-weighted *** 6 hours,24 hours,7 days and 3 months after modeling,the hemisphere anomalous volume ratio was significantly reduced after treatment with nerve growth factor. Hematoxylin-eosin staining,immunohistochemistry,electron microscopy and neurological function scores showed that infarct defects were slightly reduced and neurological function significantly improved after nerve growth factor *** result was consistent with diffusion-weighted MRI *** findings indicate that nerve growth factor can protect against cerebral infarction,and that the hemisphere anomalous volume ratio of diffusion-weighted MRI can be used to evaluate the therapeutic effect.

关键词： diffusion-weighted MRI nerve growth factor hemisphere anomalous volume ratio cerebral infarction treatment neuroprotection brain regeneration neural regeneration

Expression of nerve growth factor mRNA in splenic lymphocytes of bronchial asthma rats and its influencing factors

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Neural Regeneration Research 2008年第9期3卷 1000-1003页

作者： Jihong Dai Yonghong Wang Haixia He Respiration Center Children＇s Hospital Chongqing Medical University Chongqing 400014 China Department of Neurology the First Affiliated Hospital Chongqing Medical University Chongqing Key Laboratory of Neurology Chongqing 400016 China Department of Clinical Pharmacy College of Clinical Medicine Chongqing Medical University Chongqing 400016 China

BACKGROUND： Previous research has proved that nerve growth factor （NGF） participates in the onset of asthma by the induction of neurogenic inflammation. OBJECTIVE： To investigate the effect of interleukin-13 （IL-13） and interferon- γ; （IFN- γ ） on the expression of NGF mRNA in the splenic lymphocytes of bronchial asthma rats. DESIGN, TIME AND SETTING： The experiment, a completely randomized study based on cellular immunology, was performed in the Laboratory of Neurology in Chongqing Medical University and the Department of Clinical Pharmacy in College of Clinical Medicine, Chongqing Medical University （Chongqing, China） from January 2006 to April 2007. MATERIALS： Four adult male Wistar rats were used in this study. Rat IL-13, IFN- γ probe and the total RNA extraction kit were produced by Shanghai Sangon Biological Technology ＆ Services Co., Ltd （China）. The NGF ELISA kit was a product of Wuhan Boster Bioengineering Co., Ltd （China）. A Du-70 automatic UV spectrophotometer was produced by Beckman Company （USA）. METHODS： Rats were subjected to 1-mL intraperitoneal injections each containing 100 mg of ovalbumin, and were sensitized by using antigen solution, which was sensitized with 5×10^9 Bacillus pertussis and 100 mg aluminum hydroxide powder. Four rats were challenged with 1% ovalbumin using an ultrasonic nebulizer for 60 minutes to establish an asthmatic model. After rats were anesthetized, splenic lymphocytes were isolated and cultured in medium, which was supplemented with IL-13 or IFN- γ, for 0, 12, 24 or 48 hours. A parallel study was conducted with cultured splenic lymphocytes, which were divided into a control group, an IL-13 group and an IFN- γ group. Culture medium was added with different concentrations of IL-13 （10, 50, 100 U g/L） and IFN- γ; （1, 10, 50 u g/L）; 24 hours later, all samples were harvested. MAIN OUTCOME MEASURES： The expression levels of NGF mRNA were detected by reverse transcription-polymerase chain reaction. RESULTS： In the control group,

关键词： asthma nerve growth factor interleukin-13 γ -interferon

Effect of salviae miltiorrhizae and ligustrazine hydrochloride injection on axonal regeneration and nerve growth factor expression in a rat model of sciatic nerve injury

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Neural Regeneration Research 2009年第12期4卷 1002-1006页

作者： Min Ma Guijuan Zhang Yi Ma Dehui Li Hening Zhai Department of Traditional Chinese Medicine Medical College of Jinan University Guangzhou 510632 Guangdong Province China First Hospital Affiliated to Jinan University Guangzhou 510632 Guangdong Province China Institute of Bioengineering Jinan University Guangzhou 510632 Guangdong Province China

BACKGROUND： Studies have shown that both salviae miltiorrhizae and ligustrazine can promote protein expression of nerve growth factor （NGF） and regeneration of peripheral nerve. OBJECTIVE： To verify the effect of salviae miltiorrhizae and ligustrazine hydrochloride injection on axonal regeneration and NGF protein expression in a rat model of sciatic nerve injury. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Laboratory of Traditional Chinese Medicine and the Institute of Bioengineering of Jinan University from July to December 2008. MATERIALS： Salviae miltiorrhizae and ligustrazine hydrochloride injection （containing 20 mg salviae miJtiorrhizae and 100 mg ligustrazine per 100 mL injection） was provided by Guizhou Baite Pharmaceutical, China; salviae miltiorrhizae and ligustrazine decoctions （containing 1 g raw drug per 1 mL decoction） were provided by Guangzhou Baiyunshan factory for Traditional Chinese Medicine, China; rabbit-anti-rat NGF monoclonal antibody was provided by Beijing Biosynthesis Biotechnology, China. METHODS： A total of 80 healthy, male, Sprague Dawley rats were used to establish a sciatic nerve injury model via neurotomy, and were then randomly assigned to 4 groups： salviae miltiorrhizae and ligustrazine hydrochloride injection group （intraperitoneal injection of 35 mL/kg per day salviae miltiorrhizae and ligustrazine hydrochloride injection）, saIviae miltiorrhizae group （intragastric peffusion of 2 mL salviae miltiorrhizae）, ligustrazine group （intragastric peffusion of 2 mL ligustrazine）, and model group （intraperitoneal injection of 35 mL/kg per day saline）, with 20 rats in each group. Thereafter, rats in each group were then divided into 4 subgroups according to varying time points of 1, 2, 4, and 8 weeks post-surgery, with 5 rats in each subgroup. MAIN OUTCOME MEASURES： Axons were quantified using chromotrope 2R-brilliant green and silver staining combined with image analysis to calculate the axonal

关键词： nerve growth factor sciatic nerve salviae miltiorrhizae and ligustrazine hydrochloride injection axonal regeneration

Effect of Human Cytomegalovirus Infection on nerve growth factor Expression in Human Glioma U251 Cells

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Biomedical and Environmental Sciences 2009年第4期22卷 354-358页

作者： HAI-TAO WANG BIN WANG ZHI-JUN LIU ZHI-QIANG BAI LING LI HAI-YAN LIU DONG-MENG QIAN ZHI-YONG YAN XU-XIA SONG Department of Microbiology Qingdao Medical College and Key Laboratory of Medicine and Biotechnology of Qingdao Qingdao University Medical College Qingdao 266071 Shandong China Qingdao Center for Disease Control and Prevention Qingdao 266033 Shandong China LRB 270E University of Massachusetts Medical School Worcester MA. 01605 USA

Objective To explore the change of endogenic nerve growth factor （NGF） expression in human glioma cells infected with human cytomegalovirus （HCMV）. Methods U251 cells were cultured in RPMI 1640 culture medium and infected with HCMV AD 169 strain in vitro to establish a cell model of viral infection. Morphologic changes of U251 cells were observed under inverted microscope before and after infection with HCMV. Expression of NGF gene and protein of cells was detected by RT-PCR and Western blotting before and after infection with HCMV. Results The cytopathic effects of HCMV-infected cells appeared on day 5 after infection. However, differential NGF expression was evident on day 7. NGF expression was decreased significantly in U251 cells on day 7 after infection in comparison with control group （P〈0.05）. Conclusion HCMV can down-regulate endogenous NGF levels in human glioma cell line U251.

关键词： Human cytomegalovirus U251 cells nerve growth factor

Rapamycin promotes Schwann cell migration and nerve growth factor secretion

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Neural Regeneration Research 2014年第6期9卷 602-609页

作者： Fang Liu Haiwei Zhang Kaiming Zhang Xinyu Wang Shipu Li Yixia Yin Department of Orthopedics Second Hospital of Yueyang State Key Laboratory of Advanced Technology for Materials Synthesis and Processing and Biomedical Materials and Engineering Center Wuhan University of Technology

Rapamycin, similar to FKS06, can promote neural regeneration in vitro. We assumed that the mechanisms of action of rapamycin and FK506 in promoting peripheral nerve regeneration were similar. This study compared the effects of different concentrations of raparnycin and FK506 on Sc hwann cells and investigated effects and mechanisms of rapamycin on improving peripheral nerve regeneration. Results demonstrated that the lowest rapamycin concentration （1.53 nmol/L） more significantly promoted Schwann cell migration than the highest FK506 concentration （100μmol/L）. Rapamycin promoted the secretion of nerve growth factors and upregulated growth-associated protein 43 expression in Schwann cells, but did not significantly affect Schwann cell proliferation. Therefore, rapamycin has potential application in peripheral nerve regeneration therapy.

关键词： nerve regeneration peripheral nerve injury rapamycin FK506 Schwann cell cell mi-gration nerve growth factor growth-associated protein 43 NSFC grant neural regeneration

Preparation and Characterization of Poly Lactic Acid/Graphene Oxide/nerve growth factor Scaffold with Electrical Stimulation for Peripheral nerve Regeneration in vitro

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Journal of Wuhan University of Technology(Materials Science) 2020年第6期35卷 1149-1161页

作者：徐海星 LI Rui LI Yiping HE Qundi YAN Xiumei SHU Tao YANG Haixia LÜ Yifei LI Zheng XU Runtian 熊承杰许沛虎 Department of Pharmaceutical Engineering School of ChemistryChemical Engineering and Life ScienceWuhan University of TechnologyWuhan 430070China Wuhan Mafangshan Middle School Wuhan 430070China Orthopaedic Department General Hospital of Central Theater Command of PLAWuhan 430070China

A novel conductive drug-loading system was prepared by using an improved emulsion electrostatic spinning method which contained polylactic acid (PLA),graphene oxide (GO),and nerve growth factor (NGF) coated with bovine serum albumin (BSA) ***,the structure,mechanical properties,morphology and electrical conductivity of PLA/GO electro spun fiber membranes with different GO ratios were ***/GO scaffolds can exhibit superior porosity,hydrophilic and biomechanical properties when the GO incorporation rate is 0.5%.The addition of GO in the PLA/GO electro spun fiber membranes can also create appropriate pH environment for the repair of injured nerve when the GO incorporation rate is above 0.5%.Secondly,PLA/GO/BSA/Genipin/NGF particles (with a ratio of BSA/NGF=3:1) prepared by modified emulsion electro spinning method will release more NGF than PLA/GO/NGF *** addition,PLA/0.5%GO/NGF scaffold can maintain its structure stability for at least 8 weeks observed by scanning electron microscope (SEM).Moreover,the degradation of PLA/0.5%GO/NGF scaffold is consistent with its weight ***,in vitro assay confirmes that PLA/GO composite scaffold exhibits low cytotoxicity to RSC96 *** results have demonstrated that PLA/0.5%GO/NGF sustained-release drug sustained-release system with appropriate electrical stimulation (ES) can promote PC12 cell proliferation,and it can maintain its differentiation capability for at least 3 *** conclusion,PLA/0.5%GO/NGF sustained-release drug sustained-release system can maintain its biological activity for at least 3 weeks and promote cell proliferation with appropriate ES.

关键词： poly lactic acid graphene oxide nerve growth factor electrical stimulation peripheral nerve regeneration

Nuclear-targeted therapy of nerve growth factor conjugated with ^(125)I for human glioma U251 cells

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Neural Regeneration Research 2011年第34期6卷 2694-2702页

作者： Yun Feng Rongguang Shao Yong Yang Hongwei He Xiaofang Li Gulshan Elahi Department of Pharmacology School of Medical Science and Laboratory Medicine Jiangsu University Zhenjiang 212013 Jiangsu Province China Institute of Medicinal Biotechnology Peking Union Medical College Chinese Academy of Medical Sciences Beijing 100050 China

The present study investigated the nuclear transportation phenomenon of ^125I-nerve growth factor （NGF） and the DNA-damaging changes to U251 cells using microautoradiography and single cell electrophoresis. The results showed that ^125I-NGF inhibited the survival of p53 mutant U251 human glioma cell/tumor and enhanced the therapeutic effectiveness of vincristine in in vivo and in vitro models. In vitro experiments showed ^125I-NGF was transported into the nucleus and damaged the DNA in U251 cells. Moreover, ^125I-NGF locked the U251 cells in the G2 phase. Further investigation showed that ^125I-NGF decreased cyclin B1 protein levels in a dose dependent manner, but the level of cyclin B1 mRNA expression remained unchanged. ^125I-NGF increased phosphorylated Chkl, Chk2 and Cdc25c protein levels in U251 cells, but did not influence p53 and p21 protein expression. Moreover, ^125I-NGF and vincristine exhibited synergistic effects on reducing cyclin B1 protein levels. These results indicate that ^125I-NGF can provide anti-tumor effects by activating the ATM and ATR pathways through DNA damage.

关键词： nuclear-targeted therapy nerve growth factor ^125I glioma conjugate cell cycle arrest