BACKGROUND： The pharmacological actions of Panax notoginseng saponins （PNS） lie in removing free radicals, anti-inflammation and anti-oxygenation. It can also improve memory and behavior in rat models of Alzheime...BACKGROUND： The pharmacological actions of Panax notoginseng saponins （PNS） lie in removing free radicals, anti-inflammation and anti-oxygenation. It can also improve memory and behavior in rat models of Alzheimer＇s disease. OBJECTIVE： Using the Morris water maze, immunohistochemistry, real-time PCR and RT-PCR, this study aimed to measure improvement in spatial learning, memory, expression of amyloid precursor protein （App） and β -amyloid （A β ）, to investigate the mechanism of action of PNS in the treatment of AD in the senescence accelerated mouse-prone 8 （SAMP8） and compare the effects with huperzine A. DESIGN, TIME AND SETTING： A completely randomized grouping design, controlled animal experiment was performed in the Center for Research ＆ Development of New Drugs, Guangxi Traditional Chinese Medical University from July 2005 to April 2007. MATERIALS： Sixty male SAMP8 mice, aged 3 months, purchased from Tianjin Chinese Traditional Medical University of China, were divided into four groups： PNS high-dosage group, PNS low-dosage group, huperzine A group and control group. PNS was provided by Weihe Pharmaceutical Co., Ltd. （batch No.： Z53021485, Yuxi, Yunan Province, China）. Huperzine A was provided by Zhenyuan Pharmaceutical Co., Ltd. （batch No.： 20040801, Zhejiang, China）. METHODS： The high-dosage group and low-dosage group were treated with 93.50 and 23.38 mg/kg PNS respectively per day and the huperzine A group was treated with 0.038 6 mg/kg huperzine A per day, all by intragastric administration, for 8 consecutive weeks. The same volume of double distilled water was given to the control group. MAIN OUTCOME MEASURES： After drug administration, learning and memory abilities were assessed by place navigation and spatial probe tests. The recording indices consisted of escape latency （time-to-platform）, and the percentage of swimming time spent in each quadrant. The number of A β 1-40, A β 1-42 and App immunopositive neurons in the brains of SAMP8 mice was analyzed by immunohistochemistry. The mRNA content ofApp, tau, acetylcholinesterase, and synaptophysin （Syp） was tested by real time PCR and RT-PCR. RESULTS： The PCR results show that PNS can downregulate the expression of the App gene and upregulate the expression of the Syp gene in the parietal cortex and hippocampus of SAMP8 mice. The therapeutic effects of the PNS high-dosage group were greater than those of the PNS low-dosage group and the huperzine A group （P 〈 0.05）. The results of the Morris water maze and immunohistochemistry indicated that PNS can improve the capacity for spatial learning and memory in SAMP8 mice, and reduce the content of A β 1-40, A β 1-42 and expression of App in the brains of SAMP8 mice. The therapeutic effects of the PNS high-dosage group were greater than that of the PNS low-dosage group and the huperzine A group （P 〈 0.05）. CONCLUSION： These results support the hypothesis that PNS plays a therapeutic and protective role on the pathological lesions and learning dysfunction of Alzheimer＇s disease. The therapeutic effects of PNS for Alzheimer＇s disease are possibly achieved through downregulating the expression of the App gene and upregulating the expression of the Syp gene. The therapeutic effects of PNS are dose-dependent and are greater than the effect of huperzine A.展开更多
An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43...An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43（GAP-43） is closely associated with neurite outgrowth and axon regeneration during neural development. We speculate that an enriched environment can reduce damage to dopaminergic neurons by affecting the expression of GAP-43. This study is designed to test this hypothesis. Three-month-old female senescence-accelerated mouse prone 8（SAMP8） mice were housed for 3 months in an enriched environment or a standard environment. These mice were then subcutaneously injected in the abdomen with 14 mg/kg MPTP four times at 2-hour intervals. Morris water maze testing demonstrated that learning and memory abilities were better in the enriched environment group than in the standard environment group. Reverse-transcription polymerase chain reaction, immunohistochemistry and western blot assays showed that m RNA and protein levels of GAP-43 in the substantia nigra were higher after MPTP application in the enriched environment group compared with the standard environment group. These findings indicate that an enriched environment can increase GAP-43 expression in SAMP8 mice. The upregulation of GAP-43 may be a mechanism by which an enriched environment protects against MPTP-induced neuronal damage.展开更多
目的探讨p38丝裂原活化蛋白激酶(p38 MAPK)对髓样相关蛋白8(MRP8)/髓样相关蛋白14(MRP14)诱导小鼠胚胎成纤维细胞凋亡的影响。方法制备MRP8、MRP14、MRP8/14备用。分别取p38^(+/+)和p38-/-细胞,随机分为空白对照组、MRP8组、MRP14组、MR...目的探讨p38丝裂原活化蛋白激酶(p38 MAPK)对髓样相关蛋白8(MRP8)/髓样相关蛋白14(MRP14)诱导小鼠胚胎成纤维细胞凋亡的影响。方法制备MRP8、MRP14、MRP8/14备用。分别取p38^(+/+)和p38-/-细胞,随机分为空白对照组、MRP8组、MRP14组、MRP8/14组。MRP8组、MRP14组、MRP8/14组分别加入50μg/m L MRP8、MRP14和MRP8/14。空白对照组加入等体积的DMEM培养液。各组干预24、48 h,采用MTT法检测p38^(+/+)和p38-/-细胞活力。采用流式细胞术检测空白对照组及MRP8/14干预24、48 h组p38^(+/+)和p38-/-细胞凋亡率。采用MTT法检测空白对照组、MRP8/14组以及TLR4受体抑制剂TAK242或RAGE中和抗体处理的MRP8/14(分别设为MRP8/14+TAK242组、MRP8/14+RAGE组)干预24 h p38^(+/+)细胞活力。采用MTT法和流式细胞术检测空白对照组、MRP8/14组以及p38激酶抑制剂SB203580处理的MRP8/14(设为MRP8/14+SB203580组)干预24 h p38^(+/+)细胞活力和凋亡率。采用Western blotting法检测MRP8/14干预0、1、2、4、6、8 h p38^(+/+)细胞p38 MAPK磷酸化。结果 MRP8组、MRP14组、MRP8/14组干预24、48 h p38^(+/+)、p38-/-细胞活力均低于空白对照组(P均<0.05),但无论MRP8/14组干预24 h还是48 h,p38-/-细胞活力明显高于p38^(+/+)细胞(P均<0.05)。MRP8/14干预24、48 h组p38^(+/+)细胞凋亡率均高于空白对照组,且MRP8/14干预48 h组细胞凋亡率更高(P均<0.05);而MRP8/14干预24、48 h组p38-/-细胞凋亡率均未见明显变化(P均>0.05)。MRP8/14组、MRP8/14+RAGE组干预24 h p38^(+/+)细胞活力低于空白对照组、MRP8/14+TAK242组干预24 h(P均<0.05)。MRP8/14+SB203580组干预24 h p38^(+/+)细胞活力较MRP8/14组干预24 h明显升高,细胞凋亡率较MRP8/14组干预24 h明显降低(P均<0.05)。MRP8/14干预2、4、6 h p38^(+/+)细胞p38 MAPK磷酸化水平明显高于干预0、1、8 h(P均<0.05)。结论 p38 MAPK能促进MRP8/14诱导的小鼠成纤维细胞凋亡,其机制可能与TLR4-p38 MAPK信号通路激活有关。展开更多
基金the National Natural Science Foundation of China, No: 30560189
文摘BACKGROUND： The pharmacological actions of Panax notoginseng saponins （PNS） lie in removing free radicals, anti-inflammation and anti-oxygenation. It can also improve memory and behavior in rat models of Alzheimer＇s disease. OBJECTIVE： Using the Morris water maze, immunohistochemistry, real-time PCR and RT-PCR, this study aimed to measure improvement in spatial learning, memory, expression of amyloid precursor protein （App） and β -amyloid （A β ）, to investigate the mechanism of action of PNS in the treatment of AD in the senescence accelerated mouse-prone 8 （SAMP8） and compare the effects with huperzine A. DESIGN, TIME AND SETTING： A completely randomized grouping design, controlled animal experiment was performed in the Center for Research ＆ Development of New Drugs, Guangxi Traditional Chinese Medical University from July 2005 to April 2007. MATERIALS： Sixty male SAMP8 mice, aged 3 months, purchased from Tianjin Chinese Traditional Medical University of China, were divided into four groups： PNS high-dosage group, PNS low-dosage group, huperzine A group and control group. PNS was provided by Weihe Pharmaceutical Co., Ltd. （batch No.： Z53021485, Yuxi, Yunan Province, China）. Huperzine A was provided by Zhenyuan Pharmaceutical Co., Ltd. （batch No.： 20040801, Zhejiang, China）. METHODS： The high-dosage group and low-dosage group were treated with 93.50 and 23.38 mg/kg PNS respectively per day and the huperzine A group was treated with 0.038 6 mg/kg huperzine A per day, all by intragastric administration, for 8 consecutive weeks. The same volume of double distilled water was given to the control group. MAIN OUTCOME MEASURES： After drug administration, learning and memory abilities were assessed by place navigation and spatial probe tests. The recording indices consisted of escape latency （time-to-platform）, and the percentage of swimming time spent in each quadrant. The number of A β 1-40, A β 1-42 and App immunopositive neurons in the brains of SAMP8 mice was analyzed by immunohistochemistry. The mRNA content ofApp, tau, acetylcholinesterase, and synaptophysin （Syp） was tested by real time PCR and RT-PCR. RESULTS： The PCR results show that PNS can downregulate the expression of the App gene and upregulate the expression of the Syp gene in the parietal cortex and hippocampus of SAMP8 mice. The therapeutic effects of the PNS high-dosage group were greater than those of the PNS low-dosage group and the huperzine A group （P 〈 0.05）. The results of the Morris water maze and immunohistochemistry indicated that PNS can improve the capacity for spatial learning and memory in SAMP8 mice, and reduce the content of A β 1-40, A β 1-42 and expression of App in the brains of SAMP8 mice. The therapeutic effects of the PNS high-dosage group were greater than that of the PNS low-dosage group and the huperzine A group （P 〈 0.05）. CONCLUSION： These results support the hypothesis that PNS plays a therapeutic and protective role on the pathological lesions and learning dysfunction of Alzheimer＇s disease. The therapeutic effects of PNS for Alzheimer＇s disease are possibly achieved through downregulating the expression of the App gene and upregulating the expression of the Syp gene. The therapeutic effects of PNS are dose-dependent and are greater than the effect of huperzine A.
基金supported by a grant from the Health Department of Hebei Province of China,No.20120056,20140314the Funding Project for Introduced Abroad Study Personnel of Hebei Province of China,No.C2011003039
文摘An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43（GAP-43） is closely associated with neurite outgrowth and axon regeneration during neural development. We speculate that an enriched environment can reduce damage to dopaminergic neurons by affecting the expression of GAP-43. This study is designed to test this hypothesis. Three-month-old female senescence-accelerated mouse prone 8（SAMP8） mice were housed for 3 months in an enriched environment or a standard environment. These mice were then subcutaneously injected in the abdomen with 14 mg/kg MPTP four times at 2-hour intervals. Morris water maze testing demonstrated that learning and memory abilities were better in the enriched environment group than in the standard environment group. Reverse-transcription polymerase chain reaction, immunohistochemistry and western blot assays showed that m RNA and protein levels of GAP-43 in the substantia nigra were higher after MPTP application in the enriched environment group compared with the standard environment group. These findings indicate that an enriched environment can increase GAP-43 expression in SAMP8 mice. The upregulation of GAP-43 may be a mechanism by which an enriched environment protects against MPTP-induced neuronal damage.
文摘目的探讨p38丝裂原活化蛋白激酶(p38 MAPK)对髓样相关蛋白8(MRP8)/髓样相关蛋白14(MRP14)诱导小鼠胚胎成纤维细胞凋亡的影响。方法制备MRP8、MRP14、MRP8/14备用。分别取p38^(+/+)和p38-/-细胞,随机分为空白对照组、MRP8组、MRP14组、MRP8/14组。MRP8组、MRP14组、MRP8/14组分别加入50μg/m L MRP8、MRP14和MRP8/14。空白对照组加入等体积的DMEM培养液。各组干预24、48 h,采用MTT法检测p38^(+/+)和p38-/-细胞活力。采用流式细胞术检测空白对照组及MRP8/14干预24、48 h组p38^(+/+)和p38-/-细胞凋亡率。采用MTT法检测空白对照组、MRP8/14组以及TLR4受体抑制剂TAK242或RAGE中和抗体处理的MRP8/14(分别设为MRP8/14+TAK242组、MRP8/14+RAGE组)干预24 h p38^(+/+)细胞活力。采用MTT法和流式细胞术检测空白对照组、MRP8/14组以及p38激酶抑制剂SB203580处理的MRP8/14(设为MRP8/14+SB203580组)干预24 h p38^(+/+)细胞活力和凋亡率。采用Western blotting法检测MRP8/14干预0、1、2、4、6、8 h p38^(+/+)细胞p38 MAPK磷酸化。结果 MRP8组、MRP14组、MRP8/14组干预24、48 h p38^(+/+)、p38-/-细胞活力均低于空白对照组(P均<0.05),但无论MRP8/14组干预24 h还是48 h,p38-/-细胞活力明显高于p38^(+/+)细胞(P均<0.05)。MRP8/14干预24、48 h组p38^(+/+)细胞凋亡率均高于空白对照组,且MRP8/14干预48 h组细胞凋亡率更高(P均<0.05);而MRP8/14干预24、48 h组p38-/-细胞凋亡率均未见明显变化(P均>0.05)。MRP8/14组、MRP8/14+RAGE组干预24 h p38^(+/+)细胞活力低于空白对照组、MRP8/14+TAK242组干预24 h(P均<0.05)。MRP8/14+SB203580组干预24 h p38^(+/+)细胞活力较MRP8/14组干预24 h明显升高,细胞凋亡率较MRP8/14组干预24 h明显降低(P均<0.05)。MRP8/14干预2、4、6 h p38^(+/+)细胞p38 MAPK磷酸化水平明显高于干预0、1、8 h(P均<0.05)。结论 p38 MAPK能促进MRP8/14诱导的小鼠成纤维细胞凋亡,其机制可能与TLR4-p38 MAPK信号通路激活有关。
