集落刺激因子对慢性脑缺血老龄鼠海马锥体细胞及认知功能的保护作用

目的 观察粒细胞集落刺激因子(G-CSF)对慢性脑缺血老龄鼠认知障碍及海马锥体细胞的保护作用.方法 12个月龄雄性SD大鼠30只,随机分为造模组、对照组和干预组,每组10只.Morris水迷宫实验评估大鼠学习记忆功能,免疫组化及荧光染色后对海马CA1区锥体细胞、凋亡细胞计数.结果 定位航行实验显示,造模组逃逸时间明显长于对照组和干预组(P<0.01);空间探索实验显示,造模组停留于平台所在象限的时间和跨过平台区域的次数明显少于对照组和干预组(P<0.01和P<0.05).免疫组化染色显示,造模组海马CA1区NeuN阳性细胞明显少于对照组(P<0.01)和干预组(P<0.05).荧光染色显示,造模组凋亡细胞明显多于对照组(P<0.01)和干预组(P<0.05).造模组、对照组及干预组海马CA1区锥体细胞数量、凋亡细胞数量变化分别与空间记忆功能改善呈正相关、负相关.结论 慢性脑缺血可致老龄鼠学习记忆能力受损,G-CSF可通过促进海马锥体细胞增生、抑制细胞凋亡实现对认知功能障碍的改善作用.     

人粒细胞—巨噬细胞集落刺激因子转移表达及对慢性乙型肝炎细胞免…

为了探讨人ＧＭ－ＣＳＦ基因转移表达对慢性乙型肝炎患者细胞免疫功能的影响，运用携带人ＧＭ－ＣＳＦ基因的重组腺病毒转染慢性乙型肝炎患者ＰＢＭＣｓ，表明以重组腺病毒为载体的人ＧＭ－ＣＳＦ基因在转染细胞中，可持续表达１．２６±０．０６５￣２．０９±０．１１ｎｇｓ·ｍｌ＾－１·２４ｈ＾－１水平２６天左右。通过观察慢性乙型肝炎患者ＰＢＭＣｓ在ＧＭ－ＣＳＦ基因转染前后及分泌ＧＭ－ＣＳＦ细胞和慢性乙型肝炎患者ＰＢ     

集落刺激因子的研究与现状

本文简要地介绍了４种集落刺激因子（粒细胞巨噬细胞因子、粒细胞集落刺激因子、巨噬细胞集落刺激因子和白细胞介素３）的发现、分子生物学特性确定、生物活性研究和临床初步应用的现状和进展。     

比较粒细胞-巨噬细胞集落刺激因子与粒细胞集落刺激因子对骨髓抑制的疗效

目的：比较粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）与粒细胞集落刺激因子（Ｇ－ＣＳＦ）治疗因化学治疗（化疗）致骨髓抑制的疗效。方法：３９例恶性肿瘤病人随机分ＧＭ－ＣＳＦ组１８例（男性１２例，女性６例，中位年龄４２ａ）与Ｇ－ＣＳＦ组２１例（男性１３例，女性８例，中位年龄５３ａ）。因化疗后引起骨髓抑制，分别应用ＧＭ－ＣＳＦ１５０μｇ／ｄ与Ｇ－ＣＳＦ１００μｇ／ｄ，ｓｃ，ｑｄ×（２～６）ｄ。结果：２组外周血白细胞总数及中性粒细胞计数与用药前比较均上升（Ｐ＜０．０１），２组间比较无显著差异（Ｐ＞０．０５）。不良反应均少而轻。结论：ＧＭ－ＣＳＦ与Ｇ－ＣＳＦ治疗化疗所致的骨髓抑制，效果相似。     

皮下注射粒细胞集落刺激因子的新方法

目的对注射皮下注射粒细胞集落刺激因子的两种方法进行比较,选择最佳的注射方法。方法选取40例病人,分两组分别采取不同的注射方法,观察乳头处的药液存有量和注射后局部外渗发生率。结果新的注射方法乳头处存有的药液量平均值为0.04±0.01ml,常规注射方法乳头处存有的药液量为0.1±0.01ml;外渗发生率新的方法为5.6%,常规方法为23%。结论新的注射方法能够充分的发挥药物的疗效,避免了最大限度的药液损失。     

比较粒细胞—巨噬细胞集落刺激因子与粒细胞集落刺激因子对骨髓抑制的疗效

目的：比较粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）与粒细胞集落刺激因子（Ｇ－ＣＳＦ）治疗因化学治疗（化疗）致骨髓抑制的疗效。方法：３９例恶性肿瘤病人随机分ＧＭ－ＣＳＦ组１８例（男性１２例，女性６例，中位年龄４２ａ）与Ｇ－ＣＳＦ组２１例（男性１３例，女性８例，中位年龄５３ａ）。因化疗后引起骨髓抑制，分别应用ＧＭ－ＣＳＦ１５０μｇ／ｄ与Ｇ－ＣＳＦ１００μｇ／ｄ，ｓｃ，ｑｄ×（２￣６）ｄ。结果：２组     

粒细胞集落刺激因子对脂肪基质细胞增殖的影响

目的 探讨粒细胞集落刺激因子(G-CSF)对大鼠脂肪基质细胞(ADASc)增殖的影响.方法 将16只SD大鼠随机分为两组,即G-CSF组(8只)与对照组(8只).G-CSF组皮下注射G-CSF 20μg·kg-1·d-1,连用5 d;对照组皮下注射等量0.9%氯化钠注射液.每组分别于最后一次注射后6 h取大鼠腹膜后脂肪组织,分离、培养、鉴定ADASc,取两组第5代的ADASc,观测细胞生长的最大增殖倍数和倍增时间,流式细胞仪测定细胞周期分布.结果 差异贴壁法培养出的第5代ADASc细胞为高纯度细胞群,表型特征为CD44+CD105-CD31-CD45-.G-CSF组与对照组大鼠ADASc最大增殖倍数、倍增时间、细胞周期Go-1细胞比例差异有统计学意义(P<0.05).G-CSF组大鼠ADASc与对照组比较,具有更大的增殖倍数和更短的倍增时间,处于Go-1周期的细胞数目也相对要少.结论 差异贴壁法是培养ADASc一种比较好的方法;G-CSF可促进骨髓ADASc进入细胞增殖周期.     

粒细胞—巨噬细胞集落刺激因子的临床应用及…

本文阐述了作用于各种骨髓细胞增殖和成熟的粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）的临床应用情况及其副作用。     

回哞与展望——细胞集落刺激因子的临床评价

目的:回顾细胞集落刺激因子进展,借以评价其疗效及安全性.方法:采用近年来国内、外文献进行综述.结果与结论:于20世纪90年代上市的细胞集落刺激因子,作为生物工程药品中多肽类的代表,进展十分迅猛,临床应用也同步增多,近期国内、外专业文献的有关报道亦浩如烟海.大量文献证实,应用细胞集落刺激因子可克服肿瘤化疗和放疗引起的骨髓毒性,有利于大量强化治疗,缩短应用肿瘤化疗周期,并减少感染的并发症.     

Gypenoside attenuates white matter lesions induced by chronic cerebral hypoperfusion in rats

Cerebral white matter lesions (WMLs) are frequently observed in vascular dementia and Alzheimer's disease and are believed to be responsible for cognitive dysfunction. The cerebral WMLs are most likely caused by chronic cerebral hypoperfusion and can be experimentally induced by permanent bilateral common carotid artery occlusion (BCCAO) in rats. Previous studies found the involvement of oxidative stress and astrocytic activation in the cerebral WMLs of BCCAO rats. Gypenoside (GP), a pure component extracted from the Gyrostemma pentaphyllum Makino, a widely reputed medicinal plants in China, has been reported to have some neuroprotective effects via anti-oxidative stress and anti-inflammatory mechanisms. In the present study, we investigated the protective effect of GP against cerebral WMLs and the underlying mechanisms for its inhibition of cognitive decline in BCCAO rats. Adult male Sprague-Dawley rats were orally administered daily doses of 200 and 400 mg/kg GP for 33 days after BCCAO, and spatial learning and memory were assessed using the Morris water maze. Following behavioral testing, oxygen free radical levels and antioxidative capability were measured biochemically. The levels of lipid peroxidation and oxidative DNA damage were also assessed by immunohistochemical staining for 4-hydroxynonenal and 8-hydroxy-2′-deoxyguanosine, respectively. Activated astrocytes were also assessed by immunohistochemical staining and Western blotting with GFAP antibodies. The morphological changes were stained with Klüver-Barrera. Rats receiving 400 mg/kg GP per day performed significantly better in tests for spatial learning and memory than saline-treated rats. GP 400 mg/kg per day were found to markedly scavenge oxygen free radicals, enhance antioxidant abilities, decrease lipid peroxide production and oxidative DNA damage, and inhibit the astrocytic activation in corpus callosum and optic tract in BCCAO rats. However, GP 200 mg/kg per day had no significant effects. GP may have therapeutic potential for treating dementia induced by chronic cerebral hypoperfusion and further evaluation is warranted.     

慢性低灌注对大鼠基底前脑胆碱能系统及认知功能的影响

目的探讨慢性低灌注状态对大鼠基底前脑胆碱能神经系统及认知功能的影响。方法永久结扎SD系大鼠双侧颈总动脉建立大鼠前脑慢性低灌注状态模型,于1、2及4个月时观察基底前脑组织病理学变化、应用免疫组织化学方法检测基底前脑胆碱乙酰转移酶(Cholineacetyltransferase,ChAT)的表达水平、应用改良的MG-2型“Y”型迷宫检测大鼠学习和记忆能力。结果与对照组相比,术后第1、2和4个月时,基底前脑神经元和胶质细胞结构紊乱、呈片状和灶性坏死,ChAT阳性神经元及纤维显著减少(P<0.01),并随时间延长逐渐加重;大鼠全天总反应时间(Totalreactiontime,TRT)明显延长(P<0.01);并且以上两者呈显著负相关(r=-0.83、P<0.01)。结论慢性低灌注状态时,大鼠基底前脑发生缺血性病理改变,导致基底前脑胆碱能神经系统损害,从而在整体水平出现学习记忆障碍。低灌注状态时,基底前脑胆碱能神经系统损伤是认知功能障碍形成的重要机制。     

DDPH: improving cognitive deficits beyond its alpha 1-adrenoceptor antagonism in chronic cerebral hypoperfused rats

DDPH (1-(2, 6-dimethylphenoxy)-2-(3, 4-dimethoxyphenylethylamino) propane hydrochloride), a candidate drug known to be an alpha(1)-adrenoceptor antagonist, can efficiently penetrate through blood brain barrier and inhibit the contraction of vascular smooth muscle in the brain. In rats with chronic cerebral hypoperfusion after permanent bilateral carotid artery ligation, we found that DDPH treatment at 6 or 12 mg/kg per day for 30 days significantly reversed pathological changes such as glial cell proliferation and nuclei shrinkage and reduced neuronal cell loss. In vivo electrophysiological studies revealed that DDPH increased long-term potentiation that was inhibited in these animals. In water maze tests, the percentage of time spent in the target quadrant (Q3) for ischemic rats (20.17+/-2.87%) was much shorter than that for the sham rats (45.39+/-3.68%), but DDPH at 12 mg/kg increased the time (39.58+/-3.77%) spent in Q3 in ischemic rats by 96.23%. These data suggested that DDPH improved the learning and memory performance significantly in rats with ischemia induced by bilateral carotid artery ligation. DDPH also lowered the levels of malondialdehyde (MDA), which was increased in the hypoperfused rats, and enhanced the activities of superoxide dismutase (SOD) and glutathione peroxidase, which were decreased in these rats. Further more, immunohistochemistry, RT-PCR assays and Western blot study demonstrated that DDPH attenuated the decreased expression of NMDAR2B (NR2B) in cortex and hippocampal CA1 region of the rats after bilateral carotid artery ligation. Our results suggest that DDPH may have favorable effects for the subjects in cerebrovascular insufficiency state following ischemic stroke.     

Curcumin reverses impaired cognition and neuronal plasticity induced by chronic stress

Chronic stress occurs in everyday life and induces impaired spatial cognition, neuroendocrine and plasticity abnormalities. A potential therapeutic for these stress related disturbances is curcumin, derived from the curry spice turmeric. Previously we demonstrated that curcumin reversed the chronic stress-induced behavioral deficits in escape from an aversive stimulus, however the mechanism behind its beneficial effects on stress-induced learning defects and associated pathologies are unknown. This study investigated the effects of curcumin on restraint stress-induced spatial learning and memory dysfunction in a water maze task and on measures related neuroendocrine and plasticity changes. The results showed that memory deficits were reversed with curcumin in a dose dependent manner, as were stress-induced increases in serum corticosterone levels. These effects were similar to positive antidepressant imipramine. Additionally, curcumin prevented adverse changes in the dendritic morphology of CA3 pyramidal neurons in the hippocampus, as assessed by the changes in branch points and dendritic length. In primary hippocampal neurons it was shown that curcumin or imipramine protected hippocampal neurons against corticosterone-induced toxicity. Furthermore, the portion of calcium/calmodulin kinase II (CaMKII) that is activated (phosphorylated CaMKII, pCaMKII), and the glutamate receptor sub-type (NMDA_2B) expressions were increased in the presence of corticosterone. These effects were also blocked by curcumin or imipramine treatment. Thus, curcumin may be an effective therapeutic for learning and memory disturbances as was seen within these stress models, and its neuroprotective effect was mediated in part by normalizing the corticosterone response, resulting in down-regulating of the pCaMKII and glutamate receptor levels.     

Rutin protects against cognitive deficits and brain damage in rats with chronic cerebral hypoperfusion

BACKGROUND AND PURPOSE

Chronic cerebral hypoperfusion is a critical causative factor for the development of cognitive decline and dementia in the elderly, which involves many pathophysiological processes. Consequently, inhibition of several pathophysiological pathways is an attractive therapeutic strategy for this disorder. Rutin, a biologically active flavonoid, protects the brain against several insults through its antioxidant and anti-inflammatory properties, but its effect on cognitive deficits and brain damage caused by chronic cerebral hypoperfusion remains unknown. Here, we investigated the neuroprotective effect of rutin on cognitive impairments and the potential mechanisms underlying its action in rats with chronic cerebral hypoperfusion.

EXPERIMENTAL APPROACH

We used Sprague-Dawley rats with permanent bilateral common carotid artery occlusion (BCCAO), a well-established model of chronic cerebral hypoperfusion. After rutin treatment for 12 weeks, the neuroprotective effect of rutin in rats was evaluated by behavioural tests, biochemical and histopathological analyses.

KEY RESULTS

BCCAO rats showed marked cognitive deficits, which were improved by rutin treatment. Moreover, BCCAO rats exhibited central cholinergic dysfunction, oxidative damage, inflammatory responses and neuronal damage in the cerebral cortex and hippocampus, compared with sham-operated rats. All these effects were significantly alleviated by treatment with rutin.

CONCLUSION AND IMPLICATIONS

Our results provide new insights into the pharmacological actions of rutin and suggest that rutin has multi-targeted therapeutical potential on cognitive deficits associated with conditions with chronic cerebral hypoperfusion such as vascular dementia and Alzheimer''s disease.     

Muscarinic cholinergic modulation of synaptic transmission and plasticity in rat hippocampus following chronic lead exposure

The cholinergic system is believed to be associated with learning and memory functions. Lead (Pb²⁺) is a well-known neurotoxic metal that causes irreversible damage to the central nervous system (CNS). To investigate whether Pb²⁺ interferes with cholinergic modulation, we examined the effects of carbachol (CCh), a muscarinic cholinergic agonist, on synaptic transmission and plasticity in the CA1 area of the hippocampus of developmentally Pb²⁺-exposed rats. The results showed that: (1) In both control and Pb²⁺-exposed rats, 0.1 μM CCh significantly enhanced tetanus-induced long-term potentiation (LTP), while 5 μM CCh induced a reversible depression of field excitatory postsynaptic potentials (fEPSPs). However, both the enhancement of LTP and depression of fEPSPs were significantly smaller in Pb²⁺-exposed rats than in controls, suggesting that the extent of the effect of CCh on the cholinergic system was depressed by Pb²⁺. (2) In Pb²⁺-exposed rats, the enhancement of LTP induced by 0.1 μM CCh was attenuated by pirenzepine, a M₁AChR antagonist, but was not affected by methoctramine tetrahydrochloride (M-105), a M_2/4AChR antagonist. The depression of fEPSPs induced by 5 μM CCh was reduced by either pirenzepine or M-105. (3) Furthermore, paired-pulse facilitation (PPF) was not affected by 0.1 μM CCh in control and Pb²⁺-exposed rats but was increased by 5 μM CCh in either group; the increase in PPF was less pronounced in Pb²⁺-treated when compared to control rats. These results suggested that cholinergic modulation could be impaired by Pb²⁺, and this kind of impairment might occur via different mAChR subtypes. Our study delineated the effects of Pb²⁺ on muscarinic modulation, and this might be one of the underlying mechanisms by which Pb²⁺ impairs learning and memory.     

慢性间断性低氧对发育期幼鼠认知功能的影响及机制研究

目的 探讨慢性间断性低氧对发育期幼鼠认知功能的影响及其可能机制.方法 21日龄SD雄性幼鼠20只,随机均分为对照组、慢性间断性低氧(CIH)组.CIH组幼鼠在自制缺氧舱内每天缺氧8h,连续缺氧6周.利用Morris水迷宫检测幼鼠认知功能的变化;利用HE染色在光镜下观察海马神经元组织学结构变化.结果 与对照组比较,CIH组幼鼠出现认知功能障碍(P＜0.05);对照组海马神经元形态正常,CIH组海马神经元出现变性坏死.结论 CIH致发育期幼鼠认知功能障碍,其机制可能与海马神经元病理损伤有关.     

Enhancement of oligodendrocyte autophagy alleviates white matter injury and cognitive impairment induced by chronic cerebral hypoperfusion in rats

Cognitive impairment caused by chronic cerebral hypoperfusion(CCH) is associated with white matter injury(WMI), possibly through the alteration of autophagy. Here, the autophagy—lysosomal pathway(ALP) dysfunction in white matter(WM) and its relationship with cognitive impairment were investigated in rats subjected to two vessel occlusion(2VO). The results showed that cognitive impairment occurred by the 28th day after 2VO. Injury and autophagy activation of mature oligodendrocytes and neuronal a...     

醒脑静注射液对慢性脑缺血大鼠认知功能的作用

目的探讨醒脑静对慢性脑缺血大鼠认知功能及细胞间黏附分子的影响。方法利用24只成年大鼠,采用双侧颈总动脉结扎的方法制作慢性大鼠脑缺血模型,分为假手术组、慢性缺血组和醒脑静治疗组。治疗组腹腔注射醒脑静注射液,2个月后用Y-迷宫检测学习能力。用免疫组化法检测脑组织细胞间黏附分子1的表达。结果醒脑静治疗组大鼠Y迷宫错误反应次数明显少于对照组,与此相类似,细胞间黏附分子1的表达在治疗组也少于模型组。结论慢性脑缺血可影响大鼠的认知能力,而醒脑静可减轻认知功能的损害程度。     

芦丁对慢性脑低灌注大鼠海马组织神经元损伤及RhoA/ROCK信号通路的影响

目的 探讨芦丁对慢性脑低灌注大鼠海马组织神经元损伤及Ras同源基因家族成员A（RhoA）/Rho相关卷曲螺旋蛋白激酶（ROCK）信号通路的影响。方法 将SD大鼠随机分为假手术组、模型组、芦丁低剂量组（20 mg/kg）、芦丁中剂量组（40 mg/kg）、芦丁高剂量组（80 mg/kg）、RhoA抑制剂组（Rhosin hydrochloride,40 mg/kg）,每组9只。除假手术组外,其余各组大鼠均通过结扎颈动脉构建慢性脑低灌注大鼠模型,按照各组给药剂量进行干预处理,每天给药1次,持续4周。采用Morris水迷宫实验测定大鼠的认知功能和记忆能力,苏木素-伊红染色观察大鼠海马组织神经元状态,TUNEL染色检测大鼠海马组织神经元凋亡情况,透射电镜观察大鼠海马组织神经元超微结构,蛋白免疫印迹法检测大鼠海马组织凋亡相关蛋白[胱天蛋白酶-3（Caspase-3）、B淋巴细胞瘤-2相关X蛋白（Bax）]及RhoA/ROCK通路相关蛋白表达情况。结果 与假手术组相比,模型组大鼠海马组织神经元数量减少,神经元超微结构、细胞器结构严重受损,逃避潜伏期、海马组织神经元凋亡率、凋亡相关蛋白（Caspase-3、Bax）、RhoA、ROCK1、ROCK2蛋白表达显著升高（P<0.05）,穿越平台次数显著减少（P<0.05）;与模型组相比,芦丁各剂量组及RhoA抑制剂组大鼠神经元数量显著增多,神经元超微结构、细胞器受损等均得到一定的恢复,逃避潜伏期、海马组织神经元凋亡率、凋亡相关蛋白（Caspase-3、Bax）、RhoA、ROCK1、ROCK2蛋白表达显著降低（P<0.05）,穿越平台次数显著增多（P<0.05）,且芦丁各剂量组呈剂量依赖效应;芦丁高剂量组与RhoA抑制剂组上述指标比较差异均无统计学意义。结论 芦丁可能通过抑制RhoA/ROCK信号通路缓解慢性脑低灌注引起的大鼠神经元损伤。