BACKGROUND:Neuronal loss,synapse mutilation,and increasing malnourished axons are pathologically related to Alzheimer's disease.Microtubule-associated protein 2(MAP2) is of importance for neuronal,axonal,and dendr...BACKGROUND:Neuronal loss,synapse mutilation,and increasing malnourished axons are pathologically related to Alzheimer's disease.Microtubule-associated protein 2(MAP2) is of importance for neuronal,axonal,and dendritic generation,extension,and stabilization,as well as for the regulation of synaptic plasticity. OBJECTIVE:To investigate the antagonistic effects of natural-cerebrolysin-containing serum on beta amyloid protein 1-40(Aβ_(1-40))-induced neurotoxicity from the standpoints of cell proliferation, synaptogenesis,and cytoskeleton formation(MAP2 expression). DESIGN,TIME AND SETTING:A paralleled,controlled,neural cell,and molecular biology experiment was performed at the Institute of Integrated Chinese and Western Medicine, Shenzhen Hospital,Southern Medical University between February 2006 and April 2008. MATERIALS:PC12 cells,derived from the rat central nervous system,were purchased from Shanghai Institute of Cell Biology,Chinese Academy of Sciences,China.Aβ_(1-40) was provided by Sigma,USA.Natural-cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine,China.The natural-cerebrolysin was predominantly composed of Renshen (Radix Ginseng),Tianma(Rhizoma Gastrodiae),and Yixingye(Ginkgo Leaf) in a proportion of 1:2:2.Following conventional water extraction technology,an extract(1:20) was prepared.Each gram of extract equaled 20 grams of crude drug.In a total of 12 adult male New Zealand rabbits, six underwent intragastric administration of natural-cerebrolysin extract for 1 month to prepare natural-cerebrolysin-containing serum,and the remaining six rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS:An Alzheimer's disease in vitro model was induced in PC12 cells using Aβ_(1-40).The cells were incubated with varying doses of natural-cerebrolysin-containing serum(2.5%,5%,and 10%).Normal blank serum-treated PC12 cells served as a blank control group. MAIN OUTCOME MEASURES:Through the use of inverted phase contrast microscope,cell morphology and neurite growth were observed,neurite length was measured,and the percentage of neurite-positive cells was calculated.Cell proliferation rate was determined by MTT assay,and MAP 2 expression was detected by fluorescent immunocytochemistry. RESULTS:Following Aβ_(1-40) treatments,some PC12 cells were apoptotic/dying,and only a few short neurites were observed.Following interventions with natural-cerebrolysin-containing serum, the PC12 cells proliferated,there was an increased number of neurites,and neurite length was enhanced.After middle- and high-dose natural-cerebrolysin treatments,the percentage of neurite-positive cells,as well as the average length of neurites,was significantly greater than the normal blank serum-treated PC12 cells(P<0.05 or P<0.01).Compared with the blank control group,MAP2 expression in the Aβ_(1-40)-treated PC12 cells was significantly inhibited,and the cell proliferation rate was significantly decreased(P<0.01).Following incubations with natural-cerebrolysin-containing serum,MAP2 expression and cell proliferation rate in the PC12 cells were significantly increased in a dose-dependent manner,compared with treatments with blank control serum(P<0.05 or P<0.01). CONCLUSION:Natural-cerebrolysin exhibited antagonistic effects on neurotoxicity in Aβ_(1-40)- induced Alzheimer's disease in vitro models.These effects were likely related to cell proliferation and the upregulation of intracellular MAP2 expression.展开更多
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...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.展开更多
目的:观察异氟烷全麻围术期老年患者血清淀粉样蛋白-β(1-40)[Aβ(1-40)]和锌水平的变化。方法:将2007年6月至2008年3月拟开腹手术患者40例纳入研究。依年龄将患者分为2组:老年组(20例)与非老年组(20例)。老年组中男12例,女8例,平均年龄...目的:观察异氟烷全麻围术期老年患者血清淀粉样蛋白-β(1-40)[Aβ(1-40)]和锌水平的变化。方法:将2007年6月至2008年3月拟开腹手术患者40例纳入研究。依年龄将患者分为2组:老年组(20例)与非老年组(20例)。老年组中男12例,女8例,平均年龄(72.39±4.11)岁;非老年组中男11例,女9例,平均年龄(48.15±5.62)岁。患者入手术室后行麻醉诱导和气管插管,并吸入异氟烷1～1.5最低肺泡浓度(MAC)维持麻醉。麻醉前及麻醉后2、4、24 h采集患者肘静脉血3 ml,用酶联免疫吸附试验(ELISA)和生化法分别测定血清Aβ(1-40)和锌水平。结果:老年组与非老年组患者麻醉前血清Aβ(1-40)水平分别为(5.03±1.96)μg/ml和(3.92±0.46)μg/ml,差异有统计学意义(P<0.05)。老年组与非老年组患者麻醉后2、4、24 h Aβ(1-40)水平均增高,分别为(7.95±1.66)μg/ml、(7.63±1.51)μg/ml、(6.94±1.43)μg/ml与(5.15±0.72)μg/ml、(6.34±0.99)μg/ml、(4.81±0.81)μg/ml;2组麻醉前后以及麻醉后2组间各时间点血清Aβ(1-40)水平比较差异均有统计学意义(均P<0.05)。老年组与非老年组患者麻醉前血清锌水平分别为(14.36±1.38)μg/ml和(14.64±1.75)μg/ml。老年组与非老年组患者麻醉后2、4、24 h血清锌水平分别为(12.71±1.36)μg/ml、(10.64±1.83)μg/ml、(8.07±1.61)μg/ml和(13.10±1.68)μg/ml、(11.16±1.74)μg/ml、(9.40±1.99)μg/ml;2组麻醉前后及麻醉后24 h 2组间血清锌水平比较差异均有统计学意义(均P<0.05)。结论:异氟烷全麻围手术期老年患者血清淀粉样蛋白-β(1-40)水平明显升高,血清锌降低。展开更多
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 hip...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.展开更多
基金Supported by:Scientific and Technological Foundation of the National Administration of Traditional Chinese Medicine of China,No.02-03LP41the Scientific and Technological Key Project of Guangdong Province,No. 2006B35630007
文摘BACKGROUND:Neuronal loss,synapse mutilation,and increasing malnourished axons are pathologically related to Alzheimer's disease.Microtubule-associated protein 2(MAP2) is of importance for neuronal,axonal,and dendritic generation,extension,and stabilization,as well as for the regulation of synaptic plasticity. OBJECTIVE:To investigate the antagonistic effects of natural-cerebrolysin-containing serum on beta amyloid protein 1-40(Aβ_(1-40))-induced neurotoxicity from the standpoints of cell proliferation, synaptogenesis,and cytoskeleton formation(MAP2 expression). DESIGN,TIME AND SETTING:A paralleled,controlled,neural cell,and molecular biology experiment was performed at the Institute of Integrated Chinese and Western Medicine, Shenzhen Hospital,Southern Medical University between February 2006 and April 2008. MATERIALS:PC12 cells,derived from the rat central nervous system,were purchased from Shanghai Institute of Cell Biology,Chinese Academy of Sciences,China.Aβ_(1-40) was provided by Sigma,USA.Natural-cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine,China.The natural-cerebrolysin was predominantly composed of Renshen (Radix Ginseng),Tianma(Rhizoma Gastrodiae),and Yixingye(Ginkgo Leaf) in a proportion of 1:2:2.Following conventional water extraction technology,an extract(1:20) was prepared.Each gram of extract equaled 20 grams of crude drug.In a total of 12 adult male New Zealand rabbits, six underwent intragastric administration of natural-cerebrolysin extract for 1 month to prepare natural-cerebrolysin-containing serum,and the remaining six rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS:An Alzheimer's disease in vitro model was induced in PC12 cells using Aβ_(1-40).The cells were incubated with varying doses of natural-cerebrolysin-containing serum(2.5%,5%,and 10%).Normal blank serum-treated PC12 cells served as a blank control group. MAIN OUTCOME MEASURES:Through the use of inverted phase contrast microscope,cell morphology and neurite growth were observed,neurite length was measured,and the percentage of neurite-positive cells was calculated.Cell proliferation rate was determined by MTT assay,and MAP 2 expression was detected by fluorescent immunocytochemistry. RESULTS:Following Aβ_(1-40) treatments,some PC12 cells were apoptotic/dying,and only a few short neurites were observed.Following interventions with natural-cerebrolysin-containing serum, the PC12 cells proliferated,there was an increased number of neurites,and neurite length was enhanced.After middle- and high-dose natural-cerebrolysin treatments,the percentage of neurite-positive cells,as well as the average length of neurites,was significantly greater than the normal blank serum-treated PC12 cells(P<0.05 or P<0.01).Compared with the blank control group,MAP2 expression in the Aβ_(1-40)-treated PC12 cells was significantly inhibited,and the cell proliferation rate was significantly decreased(P<0.01).Following incubations with natural-cerebrolysin-containing serum,MAP2 expression and cell proliferation rate in the PC12 cells were significantly increased in a dose-dependent manner,compared with treatments with blank control serum(P<0.05 or P<0.01). CONCLUSION:Natural-cerebrolysin exhibited antagonistic effects on neurotoxicity in Aβ_(1-40)- induced Alzheimer's disease in vitro models.These effects were likely related to cell proliferation and the upregulation of intracellular MAP2 expression.
基金the National Natural Science Foundation of China, No. 30973779the National Special Planning Project for Traditional Chinese Medicine of China, No.02-03LP41the Key Program of Scientific Planning of Guangdong Province, No. 2006B35630007
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.
文摘目的:观察异氟烷全麻围术期老年患者血清淀粉样蛋白-β(1-40)[Aβ(1-40)]和锌水平的变化。方法:将2007年6月至2008年3月拟开腹手术患者40例纳入研究。依年龄将患者分为2组:老年组(20例)与非老年组(20例)。老年组中男12例,女8例,平均年龄(72.39±4.11)岁;非老年组中男11例,女9例,平均年龄(48.15±5.62)岁。患者入手术室后行麻醉诱导和气管插管,并吸入异氟烷1～1.5最低肺泡浓度(MAC)维持麻醉。麻醉前及麻醉后2、4、24 h采集患者肘静脉血3 ml,用酶联免疫吸附试验(ELISA)和生化法分别测定血清Aβ(1-40)和锌水平。结果:老年组与非老年组患者麻醉前血清Aβ(1-40)水平分别为(5.03±1.96)μg/ml和(3.92±0.46)μg/ml,差异有统计学意义(P<0.05)。老年组与非老年组患者麻醉后2、4、24 h Aβ(1-40)水平均增高,分别为(7.95±1.66)μg/ml、(7.63±1.51)μg/ml、(6.94±1.43)μg/ml与(5.15±0.72)μg/ml、(6.34±0.99)μg/ml、(4.81±0.81)μg/ml;2组麻醉前后以及麻醉后2组间各时间点血清Aβ(1-40)水平比较差异均有统计学意义(均P<0.05)。老年组与非老年组患者麻醉前血清锌水平分别为(14.36±1.38)μg/ml和(14.64±1.75)μg/ml。老年组与非老年组患者麻醉后2、4、24 h血清锌水平分别为(12.71±1.36)μg/ml、(10.64±1.83)μg/ml、(8.07±1.61)μg/ml和(13.10±1.68)μg/ml、(11.16±1.74)μg/ml、(9.40±1.99)μg/ml;2组麻醉前后及麻醉后24 h 2组间血清锌水平比较差异均有统计学意义(均P<0.05)。结论:异氟烷全麻围手术期老年患者血清淀粉样蛋白-β(1-40)水平明显升高,血清锌降低。
基金sponsored by the National Natural Science Foundation of China,No. 30973779
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.
