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 fact...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 （1 μg/mL; model group）, and 5%, 10%, and 15% serum containing natural cerebrolysin and tunicamycin （1 μ g/mL; low-, moderate-, and high-dose serum containing natural cerebrotysin 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.展开更多
To examine the effects of Cerebrolysin on the treatment of diabetic peripheral neuropathy, we first established a mouse model of type 2 diabetes mellitus by administering a high-glucose, high-fat diet and a single int...To examine the effects of Cerebrolysin on the treatment of diabetic peripheral neuropathy, we first established a mouse model of type 2 diabetes mellitus by administering a high-glucose, high-fat diet and a single intraperitoneal injection of streptozotocin. Mice defined as diabetic in this model were then treated with 1.80, 5.39 or 8.98 m L/kg of Cerebrolysin via intraperitoneal injections for 10 consecutive days. Our results demonstrated that the number, diameter and area of myelinated nerve fibers increased in the sciatic nerves of these mice after administration of Cerebrolysin. The results of several behavioral tests showed that Cerebrolysin dose-dependently increased the slope angle in the inclined plane test（indicating an improved ability to maintain body position）, prolonged tail-flick latency and foot-licking time（indicating enhanced sensitivity to thermal and chemical pain, respectively, and reduced pain thresholds）, and increased an index of sciatic nerve function in diabetic mice compared with those behavioral results in untreated diabetic mice. Taken together, the anatomical and functional results suggest that Cerebrolysin ameliorated peripheral neuropathy in a mouse model of type 2 diabetes mellitus.展开更多
Cerebrolysin is a drug consisting of low-molecular-weight neurotrophic peptides and free amino acids. Cerebrolysin has been shown to ameliorate the effects of oxidative stress, reduce apoptosis, and promote neuronal g...Cerebrolysin is a drug consisting of low-molecular-weight neurotrophic peptides and free amino acids. Cerebrolysin has been shown to ameliorate the effects of oxidative stress, reduce apoptosis, and promote neuronal growth in several degenerative and acquired central nervous system insults, including dementias, stroke, and traumatic injuries. Little is known about its therapeutic efficacy in peripheral nervous system diseases. In this study, we clinically evaluated the effects of cerebrolysin on peripheral nervous system lesions. We evaluated the clinical efficacy of cerebrolysin in six patients with the following conditions who failed to respond to conventional therapies： （1） atonic bladder due to inflammatory radiculitis; （2） paraplegia due to inflammatory radiculoneuropathy; （3） post-traumatic brachial plexopathy; （4） compressive radial nerve injury; （5） post-traumatic facial nerve paralysis; and （6） diabetic ophthalmoplegia. Our results showed that cerebrolysin was more associated with rapid neurological recovery after various peripheral nerve lesions than other therapies including steroids and supportive therapies such as vitamins and antioxidants. The present results support the therapeutic efficacy of cerebrolysin in the treatment of acquired peripheral nervous system diseases.展开更多
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 ...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 of 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 month for natural cerebrolysin-containing serum. The remaining six rabbits received intragastric 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 to five passages were co-cultured with LG-DMEM media containing 10% natural cerebrolysin. Simultaneously, MSCs cultured in/G-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 1.5x 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.展开更多
BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological product of Alzheimer＇s disease （AD）, results in neuronal endoplasmic reticulum stress （ERS）. However, the mech...BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological 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 poorly understood. OBJECTIVE： To measure expression levels of protective proteins （GRP78 and GRP94） of ER molecular partners and pro-apoptotic Caspase-12 ER membrane expression following application of traditional Chinese medicine natural cerebrolysin （NC） to treat Aβ1-40-induced ERS. DESIGN, TIME AND SETTING： A parallel-controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern Medical University between September 2006 and November 2008. MATERIALS： Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoral bone marrow. Isolation and purification of mesenchymal stem cells （MSCs） were established from the whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ1-40 was provided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese and Western 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 conventional water extraction technology, an extract （1 ： 20） was prepared. Six adult, male, New Zealand rabbits underwent intragastric administration of NC extract （0.976 g/kg per day） for 1 month to prepare NC-positive serum, and the remaining 6 rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： A total of 500 nmol/L Aβ1-40 was used to establish ERS models of primary cultured MSCs. AD cell models were incubated with different doses of NC-positive serum （2.5%, 5%, and 10%）. MSCs treated with normal blank serum served as normal blank controls. MAIN OUTCOME MEASURES： Reverse transcription-polymerase chain reaction and fluorescent immunocytochemistry were respectively used to measure mRNA and protein expression levels of GRP78, GRP94, and Caspase-12 in MSCs. RESULTS： Following Aβ1-40 exposure, mRNA and protein expression levels of GRP78 and GRP94, as well as Caspase-12, significantly increased （P 〈 0.05）, suggesting successful establishment of ERS models. Following NC-positive serum application, mRNA and protein expression levels of GRP78 and GRP94 in MSCs significantly increased （P 〈 0.05 or P 〈 0.01）. However, mRNA and protein expression levels of Caspase-12 significantly decreased （P 〈 0.05, or P 〈 0.01） compared with the ERS model group. These effects were dose-dependent. CONCLUSION： NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94 expression in MSCs in a dose-dependent manner under the state of Aβ1-40-induced ERS.展开更多
Objectives: Although people with amnestic mild cognitive impairment (aMCI) benefit from cerebrolysin treatment, some still develop dementia. The aim of the current study was to identify most informative clinical asses...Objectives: Although people with amnestic mild cognitive impairment (aMCI) benefit from cerebrolysin treatment, some still develop dementia. The aim of the current study was to identify most informative clinical assessment tests to predict the therapy efficacy in aMCI subjects treated with cerebrolysin. Methods: We studied patients with amnestic mild cognitive impairment (aMCI;n = 53) who had regular neurocognitive and clinical psychiatric assessments and were treated with cerebrolysin i.v. infusions 20 × 30 ml twice a year over three years period. Data were analyzed using non-parametric statistics, cluster and linear discriminant analyses. Results: Combined mathematical modeling enabled to predict cognitive decline from aMCI to dementia in the cerebrolysin-treated patients based on their initial neurocognitive assessment scores. We identified a “dementia risk group” with fast cognitive decline (i.e. low efficacy of the treatment). Lower baseline scores in the Mattis Dementia Rating Scale Memory subtest, Mini-Mental State Examination (MMSE), 10 word list immediate recall, and Frontal Assessment Battery tests when accompanied by higher depression score (Hamilton Depression Rating Scale) suggest poor prognosis for aMCI patients treated with cerebrolysin. Changes in scores on the MMSE, Boston naming test, Digit span forward, and Wechsler scale subtest “Categorical associations” during the treatment course are more characteristic for patients who convert to dementia than their initial scores. Conclusions: aMCI subjects treated with cerebrolysin with lower baseline cognitive functioning and subclinical depression have poor prognosis in terms of converting to dementia. Changes in the MMSE, Boston naming test, Digit span forward, and Wechsler scale subtest “Categorical associations” scores during the treatment course are more informative to identify patients who will develop dementia than their initial scores.展开更多
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, an...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 （Aβ1-40）-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. A β1-40 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 AIzheimer＇s disease in vitro model was induced in PC12 cells using Aβ1-40. 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 Aβ1-40 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 Aβ1-40-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 Aβ1-40 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： 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 th...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 extractJon 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 Wntsignaling pathways, which regulate nerve cell differentiation, development and function, as well as learning and memory; Ras, G protein- coupled 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.展开更多
Autonomic innervation of heart is abnormal in diabetes and produces altered cardiovascular parameters. Cerebrolysin is a neurotrophic factor that improves the dendritic tree and synapses in the central nerve system af...Autonomic innervation of heart is abnormal in diabetes and produces altered cardiovascular parameters. Cerebrolysin is a neurotrophic factor that improves the dendritic tree and synapses in the central nerve system after brain damage. The aim of this study was to evaluate if cerebrolysin can improve the cardiac neuropathy generated in diabetic rats. Male Sprague-Dawley rats two months old were injected with streptozotocin (70 mg/Kg/, ip). Hyperglycemia and altered cardiac rate were confirmed after eight weeks of STZ injection, and cerebrolysin treatment was started in control and diabetic rats for two months (1 ml/kg/day, ip). Body weight, heart rate, heart rate variability, arterial blood pressure, and blood glucose levels were measured. Also heart weight and levels of nitrites, NGF and VEGF were measured in left ventricle homogenates. The results show that body weight was reduced and blood glucose levels were increased significantly in diabetic rats. Cerebrolysin treatment produced no significant changes in body weight either in blood glucose level in control and diabetic rats. Cerebrolysin treatment in diabetic rats shows an improvement in the altered basal cardiac rate (306 ± 6.5 lat/min) compared to diabetic saline group (272 ± 8.9 lat/min: P < 0.05), without changes in control rats. Levels of nitrites, VEGF, and NGF in the left ventricle increased in diabetic cerebrolysin treated rats. In conclusion, the results show that cerebrolysin improves some abnormalities observed in the diabetic cardiac neuropathy in rats and suggest that could be considered an additional treatment to prevent or reduce the cardiac autonomic alterations generated in diabetes.展开更多
Schizophrenia and psychosis are psychiatric condition whose neural mechanisms are yet incompletely known, and for which pharmacological treatment is too often ineffective in a growing clinical cohort. Recently, dendri...Schizophrenia and psychosis are psychiatric condition whose neural mechanisms are yet incompletely known, and for which pharmacological treatment is too often ineffective in a growing clinical cohort. Recently, dendritic morphological changes in arborization and dendritic spine density in limbic regions has been reported in postmortem tissue from schizophrenic patients and in animal models of schizophrenia, suggesting that the use of medication improving synaptogenesis may be beneficial as additional treatment of psychotic patients. Cerebrolysin (Cbl) is a drug available for clinical with active neuropeptides fragments that mimics the action of endogenous neurotrophic factors such as BDNF, GDNF, CNTF and NGF, which improves the integrity of the neuronal circuits as well as cognitive and behavioral performance by exerting a neuroprotective effect and promoting the generation of new functional synapses. Recent work from our laboratory has shown that Cbl ameliorates synaptic and dendritic pathology in animal models of schizophrenia by increasing synaptic density and restoring neuronal cytoarchitecture. This neuroprotective effect improves the integrity of the neuronal circuits and improves cognitive and behavioral performance. Importantly, Cbl treatment seems to be safe when used in combination with neuroleptics such as risperidone. The present article analyzes the potential of Cbl in the treatment of neurodevelopmental disease, and reviews the current literature on the effects of Cbl in in vivo animal models of neurodevelopmental disorders like schizophrenia.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： 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 （1 μg/mL; model group）, and 5%, 10%, and 15% serum containing natural cerebrolysin and tunicamycin （1 μ g/mL; low-, moderate-, and high-dose serum containing natural cerebrotysin 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.
文摘To examine the effects of Cerebrolysin on the treatment of diabetic peripheral neuropathy, we first established a mouse model of type 2 diabetes mellitus by administering a high-glucose, high-fat diet and a single intraperitoneal injection of streptozotocin. Mice defined as diabetic in this model were then treated with 1.80, 5.39 or 8.98 m L/kg of Cerebrolysin via intraperitoneal injections for 10 consecutive days. Our results demonstrated that the number, diameter and area of myelinated nerve fibers increased in the sciatic nerves of these mice after administration of Cerebrolysin. The results of several behavioral tests showed that Cerebrolysin dose-dependently increased the slope angle in the inclined plane test（indicating an improved ability to maintain body position）, prolonged tail-flick latency and foot-licking time（indicating enhanced sensitivity to thermal and chemical pain, respectively, and reduced pain thresholds）, and increased an index of sciatic nerve function in diabetic mice compared with those behavioral results in untreated diabetic mice. Taken together, the anatomical and functional results suggest that Cerebrolysin ameliorated peripheral neuropathy in a mouse model of type 2 diabetes mellitus.
文摘Cerebrolysin is a drug consisting of low-molecular-weight neurotrophic peptides and free amino acids. Cerebrolysin has been shown to ameliorate the effects of oxidative stress, reduce apoptosis, and promote neuronal growth in several degenerative and acquired central nervous system insults, including dementias, stroke, and traumatic injuries. Little is known about its therapeutic efficacy in peripheral nervous system diseases. In this study, we clinically evaluated the effects of cerebrolysin on peripheral nervous system lesions. We evaluated the clinical efficacy of cerebrolysin in six patients with the following conditions who failed to respond to conventional therapies： （1） atonic bladder due to inflammatory radiculitis; （2） paraplegia due to inflammatory radiculoneuropathy; （3） post-traumatic brachial plexopathy; （4） compressive radial nerve injury; （5） post-traumatic facial nerve paralysis; and （6） diabetic ophthalmoplegia. Our results showed that cerebrolysin was more associated with rapid neurological recovery after various peripheral nerve lesions than other therapies including steroids and supportive therapies such as vitamins and antioxidants. The present results support the therapeutic efficacy of cerebrolysin in the treatment of acquired peripheral nervous system diseases.
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： 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 of 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 month for natural cerebrolysin-containing serum. The remaining six rabbits received intragastric 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 to five passages were co-cultured with LG-DMEM media containing 10% natural cerebrolysin. Simultaneously, MSCs cultured in/G-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 1.5x 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.
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 characteristic pathological 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 poorly understood. OBJECTIVE： To measure expression levels of protective proteins （GRP78 and GRP94） of ER molecular partners and pro-apoptotic Caspase-12 ER membrane expression following application of traditional Chinese medicine natural cerebrolysin （NC） to treat Aβ1-40-induced ERS. DESIGN, TIME AND SETTING： A parallel-controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern Medical University between September 2006 and November 2008. MATERIALS： Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoral bone marrow. Isolation and purification of mesenchymal stem cells （MSCs） were established from the whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ1-40 was provided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese and Western 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 conventional water extraction technology, an extract （1 ： 20） was prepared. Six adult, male, New Zealand rabbits underwent intragastric administration of NC extract （0.976 g/kg per day） for 1 month to prepare NC-positive serum, and the remaining 6 rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： A total of 500 nmol/L Aβ1-40 was used to establish ERS models of primary cultured MSCs. AD cell models were incubated with different doses of NC-positive serum （2.5%, 5%, and 10%）. MSCs treated with normal blank serum served as normal blank controls. MAIN OUTCOME MEASURES： Reverse transcription-polymerase chain reaction and fluorescent immunocytochemistry were respectively used to measure mRNA and protein expression levels of GRP78, GRP94, and Caspase-12 in MSCs. RESULTS： Following Aβ1-40 exposure, mRNA and protein expression levels of GRP78 and GRP94, as well as Caspase-12, significantly increased （P 〈 0.05）, suggesting successful establishment of ERS models. Following NC-positive serum application, mRNA and protein expression levels of GRP78 and GRP94 in MSCs significantly increased （P 〈 0.05 or P 〈 0.01）. However, mRNA and protein expression levels of Caspase-12 significantly decreased （P 〈 0.05, or P 〈 0.01） compared with the ERS model group. These effects were dose-dependent. CONCLUSION： NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94 expression in MSCs in a dose-dependent manner under the state of Aβ1-40-induced ERS.
文摘Objectives: Although people with amnestic mild cognitive impairment (aMCI) benefit from cerebrolysin treatment, some still develop dementia. The aim of the current study was to identify most informative clinical assessment tests to predict the therapy efficacy in aMCI subjects treated with cerebrolysin. Methods: We studied patients with amnestic mild cognitive impairment (aMCI;n = 53) who had regular neurocognitive and clinical psychiatric assessments and were treated with cerebrolysin i.v. infusions 20 × 30 ml twice a year over three years period. Data were analyzed using non-parametric statistics, cluster and linear discriminant analyses. Results: Combined mathematical modeling enabled to predict cognitive decline from aMCI to dementia in the cerebrolysin-treated patients based on their initial neurocognitive assessment scores. We identified a “dementia risk group” with fast cognitive decline (i.e. low efficacy of the treatment). Lower baseline scores in the Mattis Dementia Rating Scale Memory subtest, Mini-Mental State Examination (MMSE), 10 word list immediate recall, and Frontal Assessment Battery tests when accompanied by higher depression score (Hamilton Depression Rating Scale) suggest poor prognosis for aMCI patients treated with cerebrolysin. Changes in scores on the MMSE, Boston naming test, Digit span forward, and Wechsler scale subtest “Categorical associations” during the treatment course are more characteristic for patients who convert to dementia than their initial scores. Conclusions: aMCI subjects treated with cerebrolysin with lower baseline cognitive functioning and subclinical depression have poor prognosis in terms of converting to dementia. Changes in the MMSE, Boston naming test, Digit span forward, and Wechsler scale subtest “Categorical associations” scores during the treatment course are more informative to identify patients who will develop dementia than their initial scores.
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 （Aβ1-40）-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. A β1-40 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 AIzheimer＇s disease in vitro model was induced in PC12 cells using Aβ1-40. 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 Aβ1-40 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 Aβ1-40-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 Aβ1-40 induced Alzheimer＇s disease in vitro models. These effects were likely related to cell proliferation and the upregulation of intracellular MAP2 expression.
基金Scientific and Technological Foundation of the National Administration of Traditional Chinese Medicine of China,No.02-03LP41the Scientific and Techno-logical Key Project of Guangdong Province,No.2006B35630007
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 extractJon 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 Wntsignaling pathways, which regulate nerve cell differentiation, development and function, as well as learning and memory; Ras, G protein- coupled 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.
文摘Autonomic innervation of heart is abnormal in diabetes and produces altered cardiovascular parameters. Cerebrolysin is a neurotrophic factor that improves the dendritic tree and synapses in the central nerve system after brain damage. The aim of this study was to evaluate if cerebrolysin can improve the cardiac neuropathy generated in diabetic rats. Male Sprague-Dawley rats two months old were injected with streptozotocin (70 mg/Kg/, ip). Hyperglycemia and altered cardiac rate were confirmed after eight weeks of STZ injection, and cerebrolysin treatment was started in control and diabetic rats for two months (1 ml/kg/day, ip). Body weight, heart rate, heart rate variability, arterial blood pressure, and blood glucose levels were measured. Also heart weight and levels of nitrites, NGF and VEGF were measured in left ventricle homogenates. The results show that body weight was reduced and blood glucose levels were increased significantly in diabetic rats. Cerebrolysin treatment produced no significant changes in body weight either in blood glucose level in control and diabetic rats. Cerebrolysin treatment in diabetic rats shows an improvement in the altered basal cardiac rate (306 ± 6.5 lat/min) compared to diabetic saline group (272 ± 8.9 lat/min: P < 0.05), without changes in control rats. Levels of nitrites, VEGF, and NGF in the left ventricle increased in diabetic cerebrolysin treated rats. In conclusion, the results show that cerebrolysin improves some abnormalities observed in the diabetic cardiac neuropathy in rats and suggest that could be considered an additional treatment to prevent or reduce the cardiac autonomic alterations generated in diabetes.
文摘Schizophrenia and psychosis are psychiatric condition whose neural mechanisms are yet incompletely known, and for which pharmacological treatment is too often ineffective in a growing clinical cohort. Recently, dendritic morphological changes in arborization and dendritic spine density in limbic regions has been reported in postmortem tissue from schizophrenic patients and in animal models of schizophrenia, suggesting that the use of medication improving synaptogenesis may be beneficial as additional treatment of psychotic patients. Cerebrolysin (Cbl) is a drug available for clinical with active neuropeptides fragments that mimics the action of endogenous neurotrophic factors such as BDNF, GDNF, CNTF and NGF, which improves the integrity of the neuronal circuits as well as cognitive and behavioral performance by exerting a neuroprotective effect and promoting the generation of new functional synapses. Recent work from our laboratory has shown that Cbl ameliorates synaptic and dendritic pathology in animal models of schizophrenia by increasing synaptic density and restoring neuronal cytoarchitecture. This neuroprotective effect improves the integrity of the neuronal circuits and improves cognitive and behavioral performance. Importantly, Cbl treatment seems to be safe when used in combination with neuroleptics such as risperidone. The present article analyzes the potential of Cbl in the treatment of neurodevelopmental disease, and reviews the current literature on the effects of Cbl in in vivo animal models of neurodevelopmental disorders like schizophrenia.
