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| 1 | Effects of serum containing natural cerebrolysin on glucose-regulated protein 78 and CCAAT enhancer-binding protein homologous protein expression in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress显示文摘BACKGROUND:Glucose-regulated protein 78(GRP78),a marker of endoplasmic reticulum stress,can prolong cell survival.Alternatively,CCAAT enhancer-binding protein homologous protein(CHOP),a transcription factor specific for endoplasmic reticulum stress,can cause cell cycle arrest and cell apoptosis. OBJECTIVE:To study the protective effects of serum containing natural cerebrolysin on endoplasmic reticulum stress in tunicamycin-induced neuronal PC12 cells,and analyze the influence on GRP78 and CHOP expressions. DESIGN,TIME AND SETTING:A parallel controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine,Shenzhen Hospital,Southern Medical University,between March 2006 and August 2008. MATERIALS:Adult Sprague-Dawley rats were perfused with natural Cerebrolysin aqueous extract(0.185 g/kg/d) to produce serum containing natural Cerebrolysin.Physiological saline was used to produce blank serum.PC12 cell line was provided by Shanghai Institute of Cell Biology, Chinese Academy of Science.Tunicamycin was provided by Sigma(St.Louis,USA),and natural Cerebrolysin,containing ginseng,rhizoma gastrodiae,and gingko leaf(1:2:2),by Shengzhen Institute of Integrated Western and Traditional Chinese Medicine. METHODS:PC12 cells were treated with DMEM culture media containing 10%blank serum (normal control group),tunicamycin(1μg/mL;model group),and 5%,10%,and 15%serum containing natural cerebrolysin and tunicamycin(1μg/mL;low-,moderate-,and high-dose serum containing natural cerebrolysin groups),for 2 hours. MAIN OUTCOME MEASURES:PC12 cells were treated with tunicamycin for 48 hours after which apoptosis was measured using the TUNEL method to calculate apoptotic index.GRP78 expression was detected using immunocytochemistry.After 24 hours of treatment with tunicamycin,GRP78 and CHOP mRNA expressions were measured using RT-PCR. RESULTS:The apoptotic index and CHOP mRNA expression were in the model group and three cerebrolysin groups were significantly increased when compared to the normal control group(P<0.05).In contrast,GRP78 mRNA and protein expressions were significantly decreased(P<0.05). CONCLUSION:Serum containing natural cerebrolysin significantly reduced apoptosis in neuronal PC12 cells following tunicamycin-induced endoplasmic reticulum stress.These results may be related to an up-regulation of GRP78 expression and down-regulation of CHOP expression,both of which displayed dose-dependent effects. | Zhengzhi Wu Ming Li Andrew C.J. Huang O Xiuqing Jia Yinghong Li Manyin Chen | 2009 | Neural Regeneration Research2009，4，2： | 4 |
| 2 | Hollow plati- num spheres with nano-channels:synthesis and enhanced catalysis for oxygen reduction显示文摘 | Chen Haoming Liu Rushi Lo Manyin | 2008 | J Phys Chern B2008，112，20： | 1 |
| 3 | Analysis of hippocampal gene expression profile of Alzheimer's disease model rats using genome chip bioinformatics显示文摘In this study,an Alzheimer's disease model was established in rats through stereotactic injection of condensed amyloid beta 1-40 into the bilateral hippocampus,and the changes of gene expression profile in the hippocampus of rat models and sham-operated rats were compared by genome expression profiling analysis.Results showed that the expression of 50 genes was significantly up-regulated(fold change ≥ 2),while 21 genes were significantly down-regulated in the hippocampus of Alzheimer's disease model rats(fold change ≤ 0.5) compared with the sham-operation group.The differentially expressed genes are involved in many functions,such as brain nerve system development,neuronal differentiation and functional regulation,cellular growth,differentiation and apoptosis,synaptogenesis and plasticity,inflammatory and immune responses,ion channels/transporters,signal transduction,cell material/energy metabolism.Our findings indicate that several genes were abnormally expressed in the metabolic and signal transduction pathways in the hippocampus of amyloid beta 1-40-induced rat model of Alzheimer's disease,thereby affecting the hippocampal and brain functions. | Yinghong Li Zhengzhi Wu Yu Jin Anmin Wu Meiqun Cao Kehuan Sun Xiuqin Jia Manyin Chen | 2012 | Neural Regeneration Research2012，7，5： | 1 |
| 4 | Natural cerebrolysin induces neuronal differentiation in bone marrow mesenchymal stem cells显示文摘BACKGROUND:Bone marrow mesenchymal stem cells(MSCs) have been shown to differentiate into neuronal-like cells through the use of several factors,such as 2-mercaptoethanol,dimethyl sulfoxide,or monothioglycero However,these factors are not suitable for human use due to toxicity.Theoretically speaking,traditional Chinese medicine could be used as potential and safe factors. OBJECTIVE:To investigate the effect of natural cerebrolysin on neuronal-like differentiation of MSCs,based on protein and mRNA analyses. DESIGN,TIME AND SETTING:A parallel controlled,in vitro experiment was performed at the Institute or Integrated Chinese and Western Medicine,Shenzhen Hospital,Southern Medical University between June 2006 and April 2008. MATERIALS:Natural cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine,China.It primarily consisted of Renshen(Radix Ginseng),Tianma(Rhizoma Gastrodiae),and Yinxingye(Ginkgo Leaf) at a proportion of 1:2:2.Natural cerebrolysin extract (1:20) was prepared using conventional water extraction methodology.Each gram of extract equaled 20 grams of the crude drug.Twelve adult,male,New Zealand rabbits were included,six of which underwent intragastric administration of natural cerebrolysin extract(0.976 g/kg per day) for 1 mon h for natural cerebrolysin-containing serum.The remaining six rabbits received intragastrie administration of equal volumes of physiological saline for normal blank serum. METHODS:Sprague Dawley male rats,6-8 weeks old,were used to harvest tibial and femoral bone marrow.Isolation and purification of MSCs were established from the whole bone marrow by removing the non-adherent cells in primary and passage cultures.For cellular identification,MSCs from four o five passages were co-cultured with LG-DMEM media containing 10% natural cerebrolys in.Simultaneously,MSCs cultured in LG-DMEM media containing 10%blank rabbit serum served as the control group. MAIN OUTCOME MEASURES:Morphology of MSCs and neurite outgrowth during differentiation was observed under inverted phase contrast microscope.Neurite-positive cells were classified by neurite length that was longer than 1.5×the cell body diameter.Immunocytochemistry was used to identify purity of MSCs following passage,as well as expression of nidogen,neuron-specific enolase,glial fibrillary acidic protein,and microtubule-associated protein 2 following treatment with natural cerebrolysin.mRNA expression of neuron-specific enolase and glial fibrillary acidic protein was detected using semi-quantitative RT-PCR. RESULTS:After MSCs were treated with natural cerebrolysin for 3-5 hours,the cell bodies were larger,and small neurites - similar to neuronal neurites - were observed.The number of neurite-positive cells significantly increased compared with the control group(P<0.05).After MSCs were treated with natural cerebrolysin for 12 hours,most expressed nidogen, neuron-specific enolase,and microtubule-associated protein 2 at higher levels than the control group(P<0.01).No evident expression of glial fibrillary acidic protein was found(P>0.05). CONCLUSION:Natural cerebrolysin promoted neurite outgrowth and induced neuronal-like differentiation of MSCs. | Zhengzhi Wu Yinghong Li Andrew C. J. Huang Ming Li Min Yang Manyin Chen | 2009 | Neural Regeneration Research2009，4，3： | 1 |
| 5 | Tumor Necrosis Factor-α Mediates Cardiac Remodeling and Ventricular Dysfunction After Pressure Overload State显示文摘 | Mei Sun Manyin Chen Fayez Dawood Urszula Zurawska Jeff Y. Li Thomas Parker Zamaneh Kassiri Lorrie A. Kirshenbaum Malcolm Arnold Rama Khokha Peter P. Liu | 2007 | Circulation2007，，11： | 1 |
| 6 | Tumor Necrosis Factor-α Mediates Cardiac Remodeling and Ventricular Dysfunction After Pressure Overload State显示文摘 | Mei Sun Manyin Chen Fayez Dawood Urszula Zurawska Jeff Y. Li Thomas Parker Zamaneh Kassiri Lorrie A. Kirshenbaum Malcolm Arnold Rama Khokha Peter P. Liu | 2007 | Circulation2007，，11： | 1 |
| 7 | Effects of natural cerebrolysin on protective proteins and pro-apoptotic molecules in mesenchymal stem cells following beta-amyloid peptide1-40-induced endoplasmic reticulum stress显示文摘BACKGROUND: Studies have demonstrated that β-amyloid peptide (Aβ), a characteristicpathological product of Alzheimer's disease (AD), results in neuronal endoplasmic reticulum stress(ERS). However, the mechanisms of traditional Chinese medicine against ERS in AD are poorlyunderstood.OBJECTIVE: To measure expression levels of protective proteins (GRP78 and GRP94) of ERmolecular partners and pro-apoptotic Caspase-12 ER membrane expression following application oftraditional Chinese medicine natural cerebrolysin (NC) to treat Aβ_(1-40)-induced ERS.DESIGN, TIME AND SETTING: A parallel-controlled study was performed at the Institute ofIntegrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern MedicalUniversity between September 2006 and November 2008.MATERIALS: Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoralbone marrow. Isolation and purification of mesenchymal stem cells (MSCs) were established fromthe whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ_(1-40) wasprovided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese andWestern Medicine, China. NC was predominantly composed of Renshen (Radix Ginseng), Tianma(Rhizoma Gastrodiae), and Yinxingye (Ginkgo Leaf) in a proportion of 1: 2: 2. Following conventionalwater extraction technology, an extract (1 : 20) was prepared. Six adult, male, New Zealand rabbitsunderwent intragastric administration of NC extract (0.976 g/kg per day) for 1 month to prepareNC-positive serum, and the remaining 6 rabbits received intragastric administration of physiologicalsaline to prepare normal blank serum.METHODS: A total of 500 nmol/L Aβ_(1-40) was used to establish ERS models of primary culturedMSCs. AD cell models were incubated with different doses of NC-positive serum (2.5%, 5%, and10%). MSCs treated with normal blank serum served as normal blank controls.MAIN OUTCOME MEASURES: Reverse transcription-polymerase chain reaction and fluorescentimmunocytochemistry were respectively used to measure mRNA and protein expression levels ofGRP78, GRP94, and Caspase-12 in MSCs.RESULTS: Following Aβ_(1-40) exposure, mRNA and protein expression levels of GRP78 and GRP94,as well as Caspase-12, significantly increased (P < 0.05), suggesting successful establishment ofERS models. Following NC-positive serum application, mRNA and protein expression levels ofGRP78 and GRP94 in MSCs significantly increased (P < 0.05 or P < 0.01). However, mRNA andprotein expression levels of Caspase-12 significantly decreased (P < 0.05, or P < 0.01) comparedwith the ERS model group. These effects were dose-dependent.CONCLUSION: NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94expression in MSCs in a dose-dependent manner under the state of Aβ_(1-40)-induced ERS. | Yinghong Li Zhengzhi Wu Ming Li Xiaoli Zhang Min Yang Manyin Chen Andrew C. J.Huang O | 2009 | Neural Regeneration Research2009，4，12： | 1 |
| 8 | Gene expression in rat mesenchymal stem cells following treatment with natural cerebrolysin-containing serum Validation of a whole genome microarray technique显示文摘BACKGROUND:Natural cerebrolysin (NC),a Chinese herbal drug for the treatment of Alzheimer's disease (AD),induces mesenchymal stem cell (MSC) differentiation into neuron-like cells,with low toxicity. But the mechanisms involved in NC effects on MSCs remain poorly understood. OBJECTIVE:We used a whole genome microarray technique to further investigate the molecular,genetic,and pharmacodynamic mechanisms of NC on MSC gene expression profiles. DESIGN,TIME AND SETTING:A parallel,controlled,in vitro experiment was performed at the First Affiliated Hospital of Shenzhen University,Shenzhen Institute of Integrated Chinese and Western Medicine,China,between September 2006 and October 2008. MATERIALS:NC was provided by Shenzhen Institute of Integrated Chinese and Western Medicine,China. It was predominantly composed of Renshen (Radix Ginseng),Tianma (Rhizoma Gastrodiae),and Yinxingye (Ginkgo Leaf) and prepared by conventional water extraction technology. Twelve adult,male,New Zealand rabbits were included,six of which underwent intragastric administration of NC extract for 1 month to create NC-containing serum. METHODS:Bone marrow was collected from the tibia and femur of Sprague Dawley rats,aged 6-8 months old. Rat MSCs were isolated and purified by the whole bone marrow adherence method. After in vitro culture,MSCs from passage 4 were treated with NC-containing serum for 48 hours,and total RNA was extracted. Gene expression in MSCs was analyzed using Affymetrix whole genome microarray analysis. MAIN OUTCOME MEASURES:Differentially expressed genes in NC serum-treated MSCs. RESULTS:NC treated MSCs displayed 46 differentially expressed genes,22 with upregulated expression (fold change > 2) and 24 with downregulated expression (fold change < -2). Differentially expressed genes participated in neuronal growth,differentiation,and function,cell growth,differentiation,proliferation,apoptosis,signal transduction,substance/energy metabolism,ion transport,and immune responses. NC treatment changed levels of transforming growth factor β/bone morphogenetic proteins,Hedgehog,Bmp,and Wnt signaling pathways,which regulate nerve cell differentiation,development and function,as well as learning and memory; Ras,G proteincoupled receptor signal pathways that are related to cell growth,proliferation,and apoptosis; and mitogen-activated protein kinase kinase kinase signaling cascades. CONCLUSION:NC can regulate gene expression for many signal transduction pathways related to nerve cell differentiation,development and function,learning and memory function,as well as regulation of cell growth,differentiation,proliferation,or apoptosis to mediate the genetic effects of NC treatment on AD. | Yinghong Li Zhengzhi Wu Ming Li Xiuqin Jia Min Yang Manyin Chen | 2010 | Neural Regeneration Research2010，5，6： | 0 |
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