BACKGROUND： The present study analyzed the effect of 3 days （2 h/d） intrauterine hypoxia on learning and memory in juvenile rats, as well as the therapeutic effects of Angelica sinensis on dentate gyrus neurons, as...BACKGROUND： The present study analyzed the effect of 3 days （2 h/d） intrauterine hypoxia on learning and memory in juvenile rats, as well as the therapeutic effects of Angelica sinensis on dentate gyrus neurons, as well as learning and memory. OBJECTIVE： To explore the effects of intrauterine hypoxia on hippocampal dentate gyrus neurons, as well as learning and memory, in juvenile rats; to explore N-methyI-D-aspartate receptor-1 （NMDAR1） expression in the dentate gyrus of neonatal rats following intrauterine hypoxia, as well as prolonged hypoxia; to investigate the regulatory mechanisms of Angelica sinensis. DESIGN, TIME AND SETTING： A randomized and controlled experiment based on developmental neurobiology was performed at the Department of Histology and Embryology in Luzhou Medical College from October 2007 to October 2008. MATERIALS： Angelica sinensis solution （250 g/L） was obtained from Central South Hospital of Wuhan University, China. Neuron-specific enolase and NMDAR1 mRNA in situ hybridization reagents were provided by Wuhan Boster Biological Technology, China. Image-Pro Plus 6.0 analysis system was purchased from Media Cybernetics, USA. METHODS： Healthy pregnant Sprague Dawley rats （n = 30） were randomly divided into control （n = 10）, hypoxia （n = 10）, and Angelica （n = 10） groups. The Angelica and hypoxia pregnant rats were placed in a three-gas incubator （oxygen concentration： 13%） starting with day 14 of pregnancy for 2 hours/day for 5 consecutive days to establish a fetal rat intrauterine hypoxia model. One hour prior to modeling, the pregnant rats from the Angelica and hypoxia groups received Angelica sinensis and normal saline （8 mL/kg） injections, respectively, through the caudal vein. The control group procedures were identical to the hypoxia group, but lacked the hypoxic conditions. MAIN OUTCOME MEASURES： Brain tissues of neonatal rats were used to detect expression of NMDAR1 mRNA, and brain tissues of juvenile rats aged 30 days were used to determine neuron-specific enolase mRNA expression by in situ hybridization. Microscopic images （400x） of the hippocampal dentate gyrus were collected. The integral optical density （IOD） value of positive NMDAR1 mRNA cells in the dentate gyrus of neonatal rats, as well as the quantity and the IOD value of positive neuron-specific enolase mRNA cells in the dentate gyrus of juvenile rats, were analyzed with Image-Pro IPP6.0 software. At 30 days after birth, learning and memory parameters were measured in the juvenile rats using Morris water maze. RESULTS： The quantity and the IOD value of positive neuron-specific enolase mRNA cells in the dentate gyrus of the hypoxia group juvenile rats were significantly less than the control group （P 〈 0.05）, and also less than the Angelica group （P 〈 0.05）. The IOD value of positive NMDAR1 mRNA cells in the dentate gyrus of the hypoxia group neonatal rats was significantly greater than the control group, and also greater than the Angelica group （P 〈 0.05）. In the Morris water maze, the searching time during the probe trial and reversal probe trial was shorter in the hypoxia group juvenile rats compared with the control group, and the Angelica group was prolonged compared with the hypoxia group （P 〈 0.05）. CONCLUSION： Intrauterine hypoxia increased expression of NMDAR1 mRNA in the dentate gyrus of neonatal rats, reduced the number of dentate gyrus neurons, and negatively affected learning and memory in juvenile rats. In contrast, Angelica sinensis injection improved the intrauterine hypoxic condition, increased the number of dentate gyrus neurons, and improved the learning and memory deficits of the juvenile rats.展开更多
Ophiocordyceps sinensis has been used as one of the most valuable traditional Chinese Medication. This fungus parasitizes larva of Hepialus armoricanus, and converts each larva into a sclerotium form, in which the fru...Ophiocordyceps sinensis has been used as one of the most valuable traditional Chinese Medication. This fungus parasitizes larva of Hepialus armoricanus, and converts each larva into a sclerotium form, in which the fruit body grows. Due to the geographical limitation, where O. sinensis can only be found in Himalayas region, the natural resources are limited and very expensive. This research aims to compare the growth-rate of O. sinensis mycelia with different ingredients mix with agar media using one-factor-at-a-time method. This research demonstrated the mycelial growth-rate with different carbon sources, including monosaccharide (Fructose, Glucose), disaccharide (Maltose, Sucrose), and polysaccharide (Dextrin, Malt extract), complex organic nitrogen sources, including beef extract, yeast extract, whey protein, and soy protein, and eight different carbon to nitrogen ratios. The objective of this research is to find out the suitable carbon and organic complex nitrogen sources and ratio for the O. sinensis solid cultivation. As results, O. sinensis grew best with disaccharides comparing to the other types of carbon sources. Furthermore, O. sinensis preferred whey protein in contrast to other organic complex nitrogen sources. As for the carbon to nitrogen ratios, an optimal ratio of 18:1 was observed. Based on those experiments, carbon source shows a greater influence for the mycelial growth. Hence many different types of grains and cereals would be great candidates as the main ingredients for the O. sinensis solid cultivation.展开更多
目的探讨香椿子正丁醇提取物(n-butyl alcohol extract of toona sinensis,NBAE)对高糖引起的肾小球内皮细胞氧化应激的影响及机制。方法分别以高糖(high glucose,HG)、HG+NBAE刺激人肾小球内皮细胞,采用DCFDA法检测细胞内活性氧(reacti...目的探讨香椿子正丁醇提取物(n-butyl alcohol extract of toona sinensis,NBAE)对高糖引起的肾小球内皮细胞氧化应激的影响及机制。方法分别以高糖(high glucose,HG)、HG+NBAE刺激人肾小球内皮细胞,采用DCFDA法检测细胞内活性氧(reactive oxygenspecies,ROS)生成情况,Nitrate/Nitrite Colorimetric法检测细胞内NO的含量,Western blot检测Nrf2、NQO1及HO-1的蛋白表达,免疫荧光方法检测Nrf2、P47phox在细胞内的定位及表达。结果高糖刺激肾小球内皮细胞后出现氧化应激损伤,ROS升高,NO减少,p47phox表达量增高,Nrf2及下游蛋白NQO1、HO-1表达减少;NBAE可明显提高Nrf2的表达,并增加下游蛋白NQO1和HO-1的表达,从而抑制高糖导致的ROS升高,抑制p47phox的表达,并稳定NO的含量。结论NBAE通过激活Nrf2及其下游靶蛋白来改善高糖对肾小球内皮细胞造成的氧化应激损伤。展开更多
基金Supported by:a grant by Sichuan Provincial Science and Technology Bureau,No. 05JY029-103:a grant by Sichuan Provincial Education Bureau.No.2006A152
文摘BACKGROUND： The present study analyzed the effect of 3 days （2 h/d） intrauterine hypoxia on learning and memory in juvenile rats, as well as the therapeutic effects of Angelica sinensis on dentate gyrus neurons, as well as learning and memory. OBJECTIVE： To explore the effects of intrauterine hypoxia on hippocampal dentate gyrus neurons, as well as learning and memory, in juvenile rats; to explore N-methyI-D-aspartate receptor-1 （NMDAR1） expression in the dentate gyrus of neonatal rats following intrauterine hypoxia, as well as prolonged hypoxia; to investigate the regulatory mechanisms of Angelica sinensis. DESIGN, TIME AND SETTING： A randomized and controlled experiment based on developmental neurobiology was performed at the Department of Histology and Embryology in Luzhou Medical College from October 2007 to October 2008. MATERIALS： Angelica sinensis solution （250 g/L） was obtained from Central South Hospital of Wuhan University, China. Neuron-specific enolase and NMDAR1 mRNA in situ hybridization reagents were provided by Wuhan Boster Biological Technology, China. Image-Pro Plus 6.0 analysis system was purchased from Media Cybernetics, USA. METHODS： Healthy pregnant Sprague Dawley rats （n = 30） were randomly divided into control （n = 10）, hypoxia （n = 10）, and Angelica （n = 10） groups. The Angelica and hypoxia pregnant rats were placed in a three-gas incubator （oxygen concentration： 13%） starting with day 14 of pregnancy for 2 hours/day for 5 consecutive days to establish a fetal rat intrauterine hypoxia model. One hour prior to modeling, the pregnant rats from the Angelica and hypoxia groups received Angelica sinensis and normal saline （8 mL/kg） injections, respectively, through the caudal vein. The control group procedures were identical to the hypoxia group, but lacked the hypoxic conditions. MAIN OUTCOME MEASURES： Brain tissues of neonatal rats were used to detect expression of NMDAR1 mRNA, and brain tissues of juvenile rats aged 30 days were used to determine neuron-specific enolase mRNA expression by in situ hybridization. Microscopic images （400x） of the hippocampal dentate gyrus were collected. The integral optical density （IOD） value of positive NMDAR1 mRNA cells in the dentate gyrus of neonatal rats, as well as the quantity and the IOD value of positive neuron-specific enolase mRNA cells in the dentate gyrus of juvenile rats, were analyzed with Image-Pro IPP6.0 software. At 30 days after birth, learning and memory parameters were measured in the juvenile rats using Morris water maze. RESULTS： The quantity and the IOD value of positive neuron-specific enolase mRNA cells in the dentate gyrus of the hypoxia group juvenile rats were significantly less than the control group （P 〈 0.05）, and also less than the Angelica group （P 〈 0.05）. The IOD value of positive NMDAR1 mRNA cells in the dentate gyrus of the hypoxia group neonatal rats was significantly greater than the control group, and also greater than the Angelica group （P 〈 0.05）. In the Morris water maze, the searching time during the probe trial and reversal probe trial was shorter in the hypoxia group juvenile rats compared with the control group, and the Angelica group was prolonged compared with the hypoxia group （P 〈 0.05）. CONCLUSION： Intrauterine hypoxia increased expression of NMDAR1 mRNA in the dentate gyrus of neonatal rats, reduced the number of dentate gyrus neurons, and negatively affected learning and memory in juvenile rats. In contrast, Angelica sinensis injection improved the intrauterine hypoxic condition, increased the number of dentate gyrus neurons, and improved the learning and memory deficits of the juvenile rats.
文摘Ophiocordyceps sinensis has been used as one of the most valuable traditional Chinese Medication. This fungus parasitizes larva of Hepialus armoricanus, and converts each larva into a sclerotium form, in which the fruit body grows. Due to the geographical limitation, where O. sinensis can only be found in Himalayas region, the natural resources are limited and very expensive. This research aims to compare the growth-rate of O. sinensis mycelia with different ingredients mix with agar media using one-factor-at-a-time method. This research demonstrated the mycelial growth-rate with different carbon sources, including monosaccharide (Fructose, Glucose), disaccharide (Maltose, Sucrose), and polysaccharide (Dextrin, Malt extract), complex organic nitrogen sources, including beef extract, yeast extract, whey protein, and soy protein, and eight different carbon to nitrogen ratios. The objective of this research is to find out the suitable carbon and organic complex nitrogen sources and ratio for the O. sinensis solid cultivation. As results, O. sinensis grew best with disaccharides comparing to the other types of carbon sources. Furthermore, O. sinensis preferred whey protein in contrast to other organic complex nitrogen sources. As for the carbon to nitrogen ratios, an optimal ratio of 18:1 was observed. Based on those experiments, carbon source shows a greater influence for the mycelial growth. Hence many different types of grains and cereals would be great candidates as the main ingredients for the O. sinensis solid cultivation.
文摘目的探讨香椿子正丁醇提取物(n-butyl alcohol extract of toona sinensis,NBAE)对高糖引起的肾小球内皮细胞氧化应激的影响及机制。方法分别以高糖(high glucose,HG)、HG+NBAE刺激人肾小球内皮细胞,采用DCFDA法检测细胞内活性氧(reactive oxygenspecies,ROS)生成情况,Nitrate/Nitrite Colorimetric法检测细胞内NO的含量,Western blot检测Nrf2、NQO1及HO-1的蛋白表达,免疫荧光方法检测Nrf2、P47phox在细胞内的定位及表达。结果高糖刺激肾小球内皮细胞后出现氧化应激损伤,ROS升高,NO减少,p47phox表达量增高,Nrf2及下游蛋白NQO1、HO-1表达减少;NBAE可明显提高Nrf2的表达,并增加下游蛋白NQO1和HO-1的表达,从而抑制高糖导致的ROS升高,抑制p47phox的表达,并稳定NO的含量。结论NBAE通过激活Nrf2及其下游靶蛋白来改善高糖对肾小球内皮细胞造成的氧化应激损伤。
