The present study induced in vitro-cultured passage 4 bone marrow-derived mesenchymal stem cells to differentiate into neural-like cells with a mixture of alkaloid, polysaccharide, aglycone, glycoside, essential oils,...The present study induced in vitro-cultured passage 4 bone marrow-derived mesenchymal stem cells to differentiate into neural-like cells with a mixture of alkaloid, polysaccharide, aglycone, glycoside, essential oils, and effective components of Buyang Huanwu decoction （active principle region of decoction for invigorating yang for recuperation）. After 28 days, nestin and neuron-specific enolase were expressed in the cytoplasm. Reverse transcription-PCR and western blot analyses showed that nestin and neuron-specific enolase mRNA and protein expression was greater in the active principle region group compared with the original formula group. Results demonstrated that the active principle region of Buyang Huanwu decoction induced greater differentiation of rat bone marrow-derived mesenchymal stem cells into neural-like cells in vitro than the original Buyang Huanwu decoction formula.展开更多
BACKGROUND： The mobilization of endogenous stem cells is an effective way to promote repair following ischemic brain damage. Buyang Huanwu decoction （BHD） can effectively improve cerebral blood flow and protect aga...BACKGROUND： The mobilization of endogenous stem cells is an effective way to promote repair following ischemic brain damage. Buyang Huanwu decoction （BHD） can effectively improve cerebral blood flow and protect against cerebral ischemia/reperfusion damage. OBJECTIVE： To study the effects of BHD on cell proliferation and differentiation in the hippocampal dentate gyrus of rats following cerebral infarction, to investigate the protective effects of BHD against cerebral infarction, and to analyze the dose-effect relationship. DESIGN, TIME AND SETTING： This randomized, controlled, animal study was performed at the Laboratory of Department of Physiology, Henan College of Traditional Chinese Medicine, China from June 2007 to February 2008. MATERIALS： A total of 36 male, Sprague Dawley rats, aged 20-21 months, were equally and randomly assigned to the following groups： sham operation, model control, and nimodipine, as well as high-dose, moderate-dose, and low-dose BHD. BHD was composed of milkvetch root, Chinese angelica, red peony root, earthworm, peach seed, safflower, and Szechwan Iovage rhizome, which were provided by the Outpatient Department, Henan College of Traditional Chinese Medicine, China. METHODS： The Chinese medicinal ingredients described above were decocted. The external carotid artery was ligated in rats from the sham operation group. Rat models of focal cerebral infarction were established by middle cerebral artery occlusion in the model control and nimodipine groups, as well as the high-dose, moderate-dose, and low-dose BHD groups. The drugs were administered by gavage 5 days, as well as 2 hours, prior to model induction. Rats in the nimodipine group were daily administered a 6 mg/kg nimodipine suspension by gavage. Rats in the high-dose, moderate-dose, and low-dose BHD groups were administered daily 26, 13, and 6.5 g/kg BHD, respectively. Rats in the sham operation and model control groups were treated with an equal volume of saline. MAIN OUTCOME MEASURES： The effects of BHD on neurological dysfunction score, brain water content, cell proliferation and differentiation in the hippocampal dentate gyrus, and pathological changes in the ischemic brain hemisphere were measured in cerebral infarction rats. RESULTS： Compared with the sham operation group, the neurological dysfunction score, brain water content, number of BrdU-positive cells, BrdU/NeuN-positive cells, and BrdU/GFAP-positive cells in the hippocampal dentate gyrus significantly increased in the model control group （P 〈 0.01 ）. Compared with the model control group, neurological dysfunction score and brain water content were significantly decreased （P 〈 0.01 or 0.05）, as were the number of BrdU-positive and BrdU/NeuN-positive cells （P 〈 0.01 or 0.05）. The number of BrdU/GFAP-positive cells was significantly reduced （P 〈 0.05） in the nimodipine group, high-dose, moderate-dose, and low-dose BHD groups. Compared with the nimodipine group, the neurological dysfunction score was significantly reduced in the moderate-dose BHD group （P 〈 0.05）. However, the number of BrdU-positive cells was significantly increased in the rat hippocampal dentate gyrus in the high-dose and moderate-dose BHD groups （P 〈 0.01 or 0.05）. The following was determined by microscopy： slightly disarranged neural cells, mild vascular dilatation, inflammatory cell infiltration, and light tissue edema were observed in the nimodipine group; inflammatory celt infiltration was reduced in the low-dose BHD group; cerebral edema and inflammatory cell infiltration were significantly reduced in the high-dose and in the moderate-dose BHD group. Electron microscopy revealed lipofuscin, slightly swollen mitochondria, and normal rough endoplasmic reticulum in the high-dose and moderate-dose BHD groups. Improvement was best in the moderate-dose BHD group. CONCLUSION： Cerebral ischemia activated proliferation of neural stem cells in the rat hippocampal dentate gyrus. The actions of BHD against cerebral ischemia/reperfusion damage correlated with proliferation and differentiation of neural stem cells in the hippocampal dentate gyrus. A moderate-dose of BHD resulted in the most effective outcome.展开更多
The traditional Chinese medicine Buyang Huanwu Decoction has been shown to improve the neu- rological function of patients with stroke. However, the precise mechanisms underlying its effect remain poorly understood. I...The traditional Chinese medicine Buyang Huanwu Decoction has been shown to improve the neu- rological function of patients with stroke. However, the precise mechanisms underlying its effect remain poorly understood. In this study, we established a rat model of cerebral ischemia by middle cerebral artery occlusion and intragastrically administered 5 g/kg Buyang Huanwu Decoction, once per day, for 1, 7, 14 and 28 days after cerebral ischemia. Immunohistochemical staining revealed a number of cells positive for the neural stem cell marker nestin in the cerebral cortex, the subven- tricular zone and the ipsilateral hippocampal dentate gyrus in rat models of cerebral ischemia. Buyang Huanwu Decoction significantly increased the number of cells positive for 5-bromodeoxyuridine （BrdU）, a cell proliferation-related marker, microtubule-associated protein-2, a marker of neuronal differentiation, and growth-associated protein 43, a marker of synaptic plasticity in the ischemic rat cerebral regions. The number of positive cells peaked at 14 and 28 days after intragastric administration of Buyang Huanwu Decoction. These findings suggest that Buyang Huanwu Decoction can promote the proliferation and differentiation of neural stem cells and en- hance synaptic plasticity in ischemic rat brain tissue.展开更多
基金supported by the Natural Science Foundation of Anhui Province (Role of substance P in electroacupuncture for focal brain ischemia in rats), No. 050431003the National Natural Science Foundation of China (Target point and signal transduction pathway of Yangxin Tongmai active principle region for BMSCs), No. 81102595
文摘The present study induced in vitro-cultured passage 4 bone marrow-derived mesenchymal stem cells to differentiate into neural-like cells with a mixture of alkaloid, polysaccharide, aglycone, glycoside, essential oils, and effective components of Buyang Huanwu decoction （active principle region of decoction for invigorating yang for recuperation）. After 28 days, nestin and neuron-specific enolase were expressed in the cytoplasm. Reverse transcription-PCR and western blot analyses showed that nestin and neuron-specific enolase mRNA and protein expression was greater in the active principle region group compared with the original formula group. Results demonstrated that the active principle region of Buyang Huanwu decoction induced greater differentiation of rat bone marrow-derived mesenchymal stem cells into neural-like cells in vitro than the original Buyang Huanwu decoction formula.
文摘BACKGROUND： The mobilization of endogenous stem cells is an effective way to promote repair following ischemic brain damage. Buyang Huanwu decoction （BHD） can effectively improve cerebral blood flow and protect against cerebral ischemia/reperfusion damage. OBJECTIVE： To study the effects of BHD on cell proliferation and differentiation in the hippocampal dentate gyrus of rats following cerebral infarction, to investigate the protective effects of BHD against cerebral infarction, and to analyze the dose-effect relationship. DESIGN, TIME AND SETTING： This randomized, controlled, animal study was performed at the Laboratory of Department of Physiology, Henan College of Traditional Chinese Medicine, China from June 2007 to February 2008. MATERIALS： A total of 36 male, Sprague Dawley rats, aged 20-21 months, were equally and randomly assigned to the following groups： sham operation, model control, and nimodipine, as well as high-dose, moderate-dose, and low-dose BHD. BHD was composed of milkvetch root, Chinese angelica, red peony root, earthworm, peach seed, safflower, and Szechwan Iovage rhizome, which were provided by the Outpatient Department, Henan College of Traditional Chinese Medicine, China. METHODS： The Chinese medicinal ingredients described above were decocted. The external carotid artery was ligated in rats from the sham operation group. Rat models of focal cerebral infarction were established by middle cerebral artery occlusion in the model control and nimodipine groups, as well as the high-dose, moderate-dose, and low-dose BHD groups. The drugs were administered by gavage 5 days, as well as 2 hours, prior to model induction. Rats in the nimodipine group were daily administered a 6 mg/kg nimodipine suspension by gavage. Rats in the high-dose, moderate-dose, and low-dose BHD groups were administered daily 26, 13, and 6.5 g/kg BHD, respectively. Rats in the sham operation and model control groups were treated with an equal volume of saline. MAIN OUTCOME MEASURES： The effects of BHD on neurological dysfunction score, brain water content, cell proliferation and differentiation in the hippocampal dentate gyrus, and pathological changes in the ischemic brain hemisphere were measured in cerebral infarction rats. RESULTS： Compared with the sham operation group, the neurological dysfunction score, brain water content, number of BrdU-positive cells, BrdU/NeuN-positive cells, and BrdU/GFAP-positive cells in the hippocampal dentate gyrus significantly increased in the model control group （P 〈 0.01 ）. Compared with the model control group, neurological dysfunction score and brain water content were significantly decreased （P 〈 0.01 or 0.05）, as were the number of BrdU-positive and BrdU/NeuN-positive cells （P 〈 0.01 or 0.05）. The number of BrdU/GFAP-positive cells was significantly reduced （P 〈 0.05） in the nimodipine group, high-dose, moderate-dose, and low-dose BHD groups. Compared with the nimodipine group, the neurological dysfunction score was significantly reduced in the moderate-dose BHD group （P 〈 0.05）. However, the number of BrdU-positive cells was significantly increased in the rat hippocampal dentate gyrus in the high-dose and moderate-dose BHD groups （P 〈 0.01 or 0.05）. The following was determined by microscopy： slightly disarranged neural cells, mild vascular dilatation, inflammatory cell infiltration, and light tissue edema were observed in the nimodipine group; inflammatory celt infiltration was reduced in the low-dose BHD group; cerebral edema and inflammatory cell infiltration were significantly reduced in the high-dose and in the moderate-dose BHD group. Electron microscopy revealed lipofuscin, slightly swollen mitochondria, and normal rough endoplasmic reticulum in the high-dose and moderate-dose BHD groups. Improvement was best in the moderate-dose BHD group. CONCLUSION： Cerebral ischemia activated proliferation of neural stem cells in the rat hippocampal dentate gyrus. The actions of BHD against cerebral ischemia/reperfusion damage correlated with proliferation and differentiation of neural stem cells in the hippocampal dentate gyrus. A moderate-dose of BHD resulted in the most effective outcome.
基金supported by grants from the National Nature Science Foundation of China,No.30873355,81072939,81273989,81202694the Foundation of Educational Commission of Hunan Province in China,No.11C0954
文摘The traditional Chinese medicine Buyang Huanwu Decoction has been shown to improve the neu- rological function of patients with stroke. However, the precise mechanisms underlying its effect remain poorly understood. In this study, we established a rat model of cerebral ischemia by middle cerebral artery occlusion and intragastrically administered 5 g/kg Buyang Huanwu Decoction, once per day, for 1, 7, 14 and 28 days after cerebral ischemia. Immunohistochemical staining revealed a number of cells positive for the neural stem cell marker nestin in the cerebral cortex, the subven- tricular zone and the ipsilateral hippocampal dentate gyrus in rat models of cerebral ischemia. Buyang Huanwu Decoction significantly increased the number of cells positive for 5-bromodeoxyuridine （BrdU）, a cell proliferation-related marker, microtubule-associated protein-2, a marker of neuronal differentiation, and growth-associated protein 43, a marker of synaptic plasticity in the ischemic rat cerebral regions. The number of positive cells peaked at 14 and 28 days after intragastric administration of Buyang Huanwu Decoction. These findings suggest that Buyang Huanwu Decoction can promote the proliferation and differentiation of neural stem cells and en- hance synaptic plasticity in ischemic rat brain tissue.
