石菖蒲α-细辛醚对Lithium-Pilocarpine癫痫模型GABA系统的调控作用

目的探讨石菖蒲α-细辛醚治疗对Lithium-Pilocar-pine模型大鼠抗癫痫作用可能的分子机制。方法建立Lithium-Pilocarpine模型,通过观察α-细辛醚治疗对Lithium-Pilocarpine模型大鼠GABA含量、GAD67表达、GABA-T活性及GABAA受体表达的影响。结果①模型组大鼠癫痫持续状态(Status epilepticus,SE)后12 h～24 h海马GABA含量及GAD67表达明显降低(P<0.01),持续至SE后72 h,之后缓慢增高;②SE后各时相脑内GABA-T活性明显增高(P<0.01),以海马区GABA-T活性增高最为突出,持续至SE后1周仍明显高于正常对照组;③SE后12 h～24 h脑内GABAAR-mRNA表达明显降低,GABAAR-mRNA表达高峰主要发生在SE后48 h～72 h;④石菖蒲α-细辛醚治疗能显著增加大鼠SE后各时相海马GABA含量,增强GAD67及GABAA-RNA表达,并持续数日以上,至SE后1周恢复正常;能降低SE后增高的GABA-T活性,其下调海马区GA-BA-T活性的作用强于额叶。结论石菖蒲α-细辛醚可能通过抑制GABA-T活性以降低GABA分解代谢,上调GAD67表达使GABA合成增加,上调GABAA受体表达以增强GA-BA介导的抑制功能来发挥抗癫痫作用。

Pilocarpine促内耳分泌功能的实验研究

目的探讨Ｐｉｌｏｃａｒｐｉｎｅ是否具有促使内耳分泌细胞功能亢进作用。方法 皮下注射Ｐｉｌｏ－ｃａｒｐｉｎｅ，用圆窗电极记录栗鼠ＣＭ，ＣＡＰ，ＳＰ，观察在用药前以及用药后３０’、６０’、９０’、１２０’、１５０’、１８０’检测栗鼠５００、１０００、２０００、４０００和８０００Ｈｚ的ＣＡＰ听阈，ＣＭ、ＳＰ振幅变化；实验结束后将动物处死，透射电镜下观察内耳形态学改变。结果皮下注射Ｐｉｌｏｃａｒｐｉｎｅ后１５’动物出现唾液分泌、排泄物增多的现象．３０’时所有频率的ＳＰ振幅增大，以 ２０００Ｈｚ最为显著，以后随着时间的延长，ＳＰ振幅逐渐增大，９０’后逐渐下降，１８０’时部分频率的ＳＰ振幅已恢复到用药前水平。在用药后各个频率的ＣＡＰ阈值的变化率呈正态分布，大多数动物无显著变化，只有少量动物的几个频率ＣＡＰ振幅在１０ｄＢ以内振荡，ＣＭ振幅无显著变化。透射电镜下内耳毛细胞形态正常，但在微纹区的暗细胞和内齿细胞（ｉｎｔｅｒｄｅｎｔａｌ ｃｅｌｌ， ＩＤＣ）内有着色较深的颗粒，并且部分颗粒向外分泌，在基底膜与盖膜间的空腔间形成层状物。结论 Ｐｉｌｏｃａｒｐｉｎｅ能促进内耳分泌功能。

Purslane protects against the reproductive toxicity of carbamazepine treatment in pilocarpine-induced epilepsy model

Objective: To investigate the protective effect of purslane with carbamazepine treatment.Methods: Male albino rats were modulated by pilocarpine to be epileptic.Both the normal and epileptic rats were treated with carbamazepine, purslane or carbamazepine plus purslane, with separate non-treated control groups for both normal and epileptic rats.Results: The data from the current study showed amelioration in amino acids and electrolytes in the epileptic rats treated with purslane and carbamazepine, with this amelioration occurring without decreasing the fertility hormones(testosterone,dehydroepiandrosterone, luteinizing hormone and follicle stimulating hormone).Purslane treatments also prevented the increase in estradiol.The decreased epileptic hyperexcitability with purslane was evidenced by decreased glial fibrillary acidic protein and lipid peroxidation.Conclusions: Natural products like purslane could be used with the highly repetitive drugs like carbamazepine to reduce or prevent its side-effects.

Correlation Study on Expression of GST-π Protein in Brain Tissue and Peripheral Blood of Epilepsy Rats Induced by Pilocarpine

Previous studies have suggested that glutathione-S-transferase π （GST-π） over-expression in the brain tissue is associated with refractory epilepsy. However, whether the change in GST-π level in the peripheral blood is in line with that in brain tissue remains unknown. This study examined the correlation between GST-π in brain tissue and that in peripheral blood in rat models of pilocarpine-induced refractory epilepsy. The animals were divided into drug-resistant group and drug-responsive group according to the response to anti-epileptic drugs. GST-π expression in brain tissue was immunohistochemically determined, while the expression of GST-π in peripheral blood was analyzed by Western blotting. In the hippocampus and cortex, GST-π was mainly found in the cytoplasm and membrane of neurons, and the GST-π expression level was higher in drug-resistant group than in the drug-responsive group and saline control group （P〈0.05）. Moreover, there was no significant difference between responders and saline control animals （P〉0.05）. The change in expression of GST-π in peripheral blood showed the same pattern as that in brain tissues, suggesting GST-π might contribute to drug resistance in epilepsy. Importantly, the GST-π over-expression in peripheral blood could be used as a marker for resistance to anti-epileptic agents.

Proliferation changes in hippocampal neurons following pilocarpine-induced intractable epilepsy in adult rats

Epilepsy can lead to the changes in neurons residing in the dentate gyrus. The present study aimed to observe the cell dividing features following epilepsy in adult rat hippocampi, and to study difference in cell proliferation between adult rats with common epilepsy and intractable epilepsy.Adult, male, Sprague Dawley rats ware randomly divided into control （n = 8, treatment with normal saline） and three expenmental groups： common epilepsy （n = 33）, intractable epilepsy （n = 11）, and drug-responsive （n = 25）. Pilocarpine （15 mg/kg） was intrapentoneally administered to establish epilepsy in the three experimental groups. Rats that developed epilepsy were treated with chloral hydrate. Rats that did not exhibit spontaneous seizures were enrolled in the common epilepsy group, and rats with spontaneous seizure were included in the spontaneous seizure group. At 6 hours after epileptic attack termination, rats ware intraperitoneally injected with bromodeoxyuridine （BrdU; 50 mg/kg）, an optimal marker forlabeling cell proliferation in vivo, four times.Immunohistochemistry results at 48 hours after BrdU injection indicated that the number of BrdU-positive cells was the highest in the common epilepsy group, followed by the control group,and lastly the intractable group （P 〈 0.01）. In addition, the number of BrdU-positive cells in the common epilepsy group was similar to the drug-responsive group. The present findings demonstrated that intractable epilepsy led to decreased hippocampal neurons in adult rats when compared to common epilepsy.

Scorpion ethanol extract and valproic acid effects on hippocampal glial fibrillary acidic protein expression in a rat model of chronic-kindling epilepsy induced by lithium chloride-pilocarpine

The present study analyzed the effects of ethanol extracts of scorpion on epilepsy prevention and hippocampal expression of glial fibrillary acidic protein in a lithium chloride-pilocarpine epileptic rat model. Results were subsequently compared with valproic acid. Results showed gradually- increased hippocampal glial fibrillary acidic protein expression following model establishment; glial fibrillary acidic protein mRNA expression was significantly increased at 3 days, reached a peak at 7 days, and then gradually decreased thereafter. Ethanol extracts of scorpion doses of 580 and 1 160 mg/kg, as well as 120 mg/kg valproic acid, led to a decreased number of glial fibrillary acidic protein-positive cells and glial fibrillary acidic protein mRNA expression, as well as decreased seizure grades and frequency of spontaneously recurrent seizures. The effects of 1 160 mg/kg ethanol extracts of scorpion were equal to those of 120 mg/kg valproic acid. These results suggested that the anti-epileptic effect of ethanol extracts of scorpion were associated with decreased hippocampal glial fibrillary acidic protein expression in a rat model of lithium chlofide-pilocarpine induced epilepsy.

Dynamic expression of P-glycoprotein in the CA1,CA3 and dentate gyrus of the rat hippocampus following lithium-pilocarpine-induced status epilepticus

Epileptic seizures induce overexpression of P-glycoprotein in the blood-brain barrier. However, it is unclear whether hippocampal neurons also overexpress P-glycoprotein following seizure. This study confirmed that the clinical manifestation, pathological characteristics and electroencephalography in the rat model of lithium-pilocarpine-induced mesial temporal lobe epilepsy were consistent with clinical reports of mesial temporal lobe epilepsy in humans.Immunohistochemistry staining demonstrated that P-glycoprotein positive staining was found in neurons in the pyramidal layer of the hippocampus. Westem blot assay and real-time polymerase chain reaction revealed that P-glycoprotein overexpression was exhibited in the CA1, CA3, and dentate gyrus of the hippocampus at 24 and 60 days following model induction, but no significant dffierence was detected in the same region at various time points. These results indicate that seizures led to overexpression of P-glycoprotein in neurons of the hippocampus, but no evidence was found for a positive association between P-glycoprotein expression and seizure frequency.

Effects of ethanol extracts of scorpion on hippocampal apoptosis and caspase-3 expression in lithium chloride-pilocarpine-induced status epilepticus rats

BACKGROUND： Previous studies have demonstrated that scorpion venom in the scorpion can inhibit epilepsy and apoptosis. However, it remains unclear whether ethanol extracts of scorpion （EES） exhibit similar effects. OBJECTIVE： To investigate the effects of EES on hippocampal apoptosis and caspase-3 expression, and to compare the effects on sodium valproate （positive control drug） in a rat model of status epilepticus induced by lithium chloride-pilocarpine. DESIGN, TIME AND SETTING： This randomized, controlled study was conducted at the Drug Research and Development Center, Kanghong Pharmaceuticals Group, and the Department of Pathology, Sichuan Academy of Medical Sciences ＆ Sichuan Provincial People＇s Hospital, China from May 2007 to April 2008. MATERIALS： EES were prepared by Huashen Pharmaceutical, China. Sodium valproate （Hunan Xiangzhong Pharmaceutical, China） and lithium chloride-pilocarpine （Sigma, USA） were also used in the present study. METHODS： From a total of 156 rats, six served as normal controls. The remaining rats were intraperitoneally injected with lithium chloride-pilocarpine to establish status epileptlcus models, and then assigned to five groups （n = 30, respectively）. Animals in each group were administered drugs at 15 minutes after epileptic seizure by gavage, i.e. in the normal control and model groups, rats were treated with 1 mL/0.1 kg saline. The sodium valproate group was administered 120 mg/kg/d sodium valproate. The low-, moderate-, and high-dose EES groups received treatments of 290, 580 and 1 160 mg/kg/d EES. The dispensed concentration was 1 mL/0.1 kg. Rat seizure behavior was observed. If status epilepticus did not terminated after 1 hour, the rats were intraperitoneally administered atropine （1 mg/kg） and diazepam （10 mg/kg） to terminate seizure. These rats were continuously observed for 6 hours to ensure seizure termination. Then rats were treated with the above-mentioned drugs at 8：00 am each day until sacrifice, which took place 4 hours after drug administration. MAIN OUTCOME MEASURES： Terminal dUTP nick end labeling （TUNEL）-positive cells and caspase-3 expression were, respectively, determined by TUNEL and immunohistochemistry at 6, 24 48, and 72 hours, as well as 7 days, after status epilepticus. Behavioral changes were also measured. RESULTS： A few caspase-3-positive cells were observed. TUNEL- and caspase-3-positive ceils were mainly visible in the hippocampal CA1 and CA3 regions 6 hours following status epilepticus in the model and drug intervention groups. The number of TUNEL-positive cells reached a peak at 48 hours following status epilepticus in the sodium valproate group, as well as the moderate- and high-dose EES groups, and number of TUNEL-positive cells reached a peak at 72 hours in the model and low-dose EES groups. The number of caspase-3-positive cells reached a peak at 48 hours in each group. Following treatment of sodium valproate and EES, the number of TUNEL- and caspase-3-positive cells significantly decreased compared with the model group at various time points （P 〈 0.05）. The number of TUNEL- and caspase-3-positive cells was greatest in the low-dose EES group, followed by the moderate- and high-dose EES groups. The number of TUNEL- and caspase-3-positive cells was similar between the sodium valproate and high-dose EES groups. Epileptic seizure was significantly improved in the sodium valproate group, as well as the moderate- and high-dose EES groups, compared with the model group （P〈 0.05 or P〈 0.01）. Treatment with sodium valproate and high-dose EES resulted in the best outcome, although the results were similar （P 〉 0.05）. CONCLUSION： A dose of 1 160 mg/kg/d EES significantly inhibited status epilepticus. This outcome corresponded to a decreased number of apoptotlc cells and caspase-3-positive cells, which was similar to sodium valproate. These results suggest that it is not necessary to extract a component from the scorpion for the treatment of epilepsy. The high dose of EES significantly inhibited epilepsy, which correlated with decreased hippocampal caspase-3 expression.

Impact of Combination Use of 0.004% Travoprost and 2% Pilocarpine on Matrix Metalloproteinases Synthesized by Rabbit Ciliary Muscle:a Pilot Study

Objective To explore the impact of combination use of prostaglandin analogue and cholinergic agonists on main matrix metalloproteinases(MMPs)synthesized by albino rabbit ciliary muscle.Methods Normal adult albino rabbits were divided into the control group,2%pilocarpine group,0.004%travoprost group and travoprost plus pilocarpine group.Two rabbits in the control group were executed after treated with normal saline for one day.Two rabbits were separately executed on the 7th,14th and 24th day of the treatment in each drug treated group.In each subgroup ciliary muscle band of 4 eyes was taken and made into homogenate.The MMPs activities of 10 subgroups were assayed by zymography.Bands’intensity which represents the activity of MMPs was measured by the UltraViolet Illumination system.Results A bright band of MMP-1/2 was showed on each lane at the position corresponding to the molecular weight of 62 kD in the ciliary smooth muscles electrophoresis.When ion Zn and Ca was displaced by MMPs inhibitor EDTA,this bright band disappeared.Compared with the control group,MMP1/2 activity increased by 4.0%,4.1%and 14.0%after 7,14 and 24 days of pilocarpine treatment.Corresponding data was23.2%,61.7%and 111.5%in the travoprost group and 49.3%,68.0%and 88.4%in the travoprost plus pilocarpine group.Conclusions Pilocarpine has little effect on activity of MMP1/2.Travoprost can increase activity of MMP1/2 gradually.Activity of MMP1/2 is rapidly increased by pilocarpine combined with travoprost,but shows small change with the prolonged treatment.

Cytotoxicity of pilocarpine to human corneal stromal cells and its underlying cytotoxic mechanisms

AIM： To examine the cytotoxic effect of pilocarpine, an anti-glaucoma drug, on human corneal stromal（HCS）cells and its underlying cytotoxic mechanisms using an in vitro model of non-transfected HCS cells.· METHODS： After HCS cells were treated with pilocarpine at a concentration from 0.15625 g/L to 20.0 g/L,their morphology and viability were detected by light microscopy and MTT assay. The membrane permeability,DNA fragmentation and ultrastructure were examined by acridine orange（AO）/ethidium bromide（EB） double-staining. DNA electrophoresis and transmission electron microscopy（TEM）, cell cycle, phosphatidylserine（PS）orientation and mitochondrial transmembrane potential（MTP） were assayed by flow cytometry（FCM）. And the activation of caspases was checked by ELISA.· RESULTS： Morphology observations and viability assay showed that pilocarpine at concentrations above0.625 g/L induced dose- and time-dependent morphological abnormality and viability decline of HCS cells. AO/EB double-staining, DNA electrophoresis and TEM noted that pilocarpine at concentrations above 0.625 g/L induced dose- and/or time-dependent membrane permeability elevation, DNA fragmentation, and apoptotic body formation of the cells. Moreover, FCM and ELISA assays revealed that 2.5 g/L pilocarpine also induced S phase arrest, PS externalization, MTP disruption, and caspase-8,-9 and-3 activation of the cells.· CONCLUSION： Pilocarpine at concentrations above0.625 g/L（1/32 of its clinical therapeutic dosage） has a dose- and time-dependent cytotoxicity to HCS cells by inducing apoptosis in these cells, which is most probably regulated by a death receptor-mediated mitochondrion-dependent signaling pathway.

A Single Hypoxic Event Ameliorates Pilocarpine Induced Hyperkinetic Movements in Planaria

Intermittent hypoxia or hypoxia therapy is exposing an individual to oxygenation conditions that are below atmospheric levels in a planned or acute timeframe. Hypoxia therapy is a potentially novel therapeutic strategy for a variety of pathologies including: mitochondrial disorders, exercise training, and mild cognitive impairments. Mitochondrial dysfunction, hyperkinetic movements, and cognitive impairments are hallmarks of seizures and status epilepticus (SE). A seizure can be considered uncontrolled electrical activity in the brain and SE is a seizure lasting more than 30 minutes, or multiple seizures without regaining consciousness in between. We examined the possibility of using the Pilocarpine model for seizure like activity on brown planaria (Dugesia tigrine). Pilocarpine is a muscarinic acetylcholine receptor agonist capable of creating seizure related brain damage. We utilized 5 mM dosages of pilocarpine and then measured open field behaviour for 3 minutes. Mobility and aversive hyperkinetic movements were observed throughout the measurement phase. After exposure to 5 mM pilocarpine, the planaria displayed behaviours consistent with seizures (e.g. aversive hyperkinetic movements and decreased mobility). Additionally, we measured the effects of an acute hypoxic event on Planaria behaviour. We used 25% carbonated water to create a hypoxic environment for the planaria and then measured mobility and hyperkinetic movements for 3 minutes. We noted that exposure to the hypoxic en-vironment produced no changes in behaviour. However, the aversive hyperkinetic move-ments produced with pilocarpine administration were completely absent when a brief (3 minutes) hypoxic episode followed the pilocarpine exposure (p < 0.05). Aversive behav-iours remained present when the ordering of pilocarpine and hypoxia were counterbal-anced. This ordering effect was consistent across 40 trials. Further evaluation of the pilo-carpine seizure model and intermittent hypoxia on planarian behaviour is warranted.

Effects of Pilocarpine Hydrochloride on the Hyposalivation Induced by Psychotropic Drugs

The aim was to study the secretagogue action of pilocarpine on the murine parotid glands submitted to chronic treatment with psychotropic drugs by salivary flow rate determinations and histological alterations. Fifty four male Wistar rats were equally divided in three groups： C group （control） received saline solution for 30 days; AD group （n = 18） received AmitriptylineR and DiazepamR for 30 days, and ADP group （n = 18） received Amitriptyline R and DiazepamR for 30 days and AmitriptylineR, DiazepamR and pilocarpine for further 30 days, resulting in 60 days of treatment. Saliva samples were collected 30 h after the end of treatment. Parotids were removed and processed for hematoxylin-eosin histological analysis. Dedicated software for image processing allowed the determination of cell number and volume. Significant differences between paired-groups C-AD （P 〈 0.01） and AD-ADP （P 〈 0.01） were observed for glands size and weight. The volume of serous cells was greater in AD, suggesting a hypertrophy of the salivary glands. For salivary flow rate, C group showed the highest average. The number of serous cells was similar between groups ADP and C, with the lowest average being found in AD group （P 〈 0.05）.

Molecular Mechanism of Bovine Trabecular Meshwork Cells Apoptosis Induced by Dexamethasone and Protection by Pilocarpine

Purpose: To study the molecular mechanism of trabecular meshwork cells apoptosis induced by dexamethasone and the protection of pilocarpine.Methods: Determining mRNA expression with reverse transcription-polymerase chain reaction (RT-PCR), protein expression with Western blots and the percentage of apoptotic cells with fluorescent microscopy.Results: Dexamethasone up-regulated Fas proteins and affected Bax, caspase-8 and caspase-9 proteins in an action of first decrease then increase. Pre-treatment with pilocarpine decreased the four proteins expression, which were increased by dexamethasone. Pilocarpine self could decrease pro-apoptotic factors Bax, caspase-8 and caspase-9 proteins expression.Conclusion: Fas/FasL pathway participated in apoptotic process induced by dexamethasone in trabecular meshwork cells and the process was probably related with both caspase-8 and caspase-9 pathways. Pilocarpine protected the cells against apoptosis through down-regulating Fas, Bax, caspase-8 and caspase-9 proteins expression.

毛果芸香碱减轻对乙酰氨基酚所致肝损伤的作用及机制研究

目的 研究毛果芸香碱减轻对乙酰氨基酚(Acetaminophen, APAP)所致肝损伤的作用及机制。方法 采用小鼠腹腔注射APAP建立急性肝损伤模型,将18只C57BL/6小鼠随机分为空白对照组、APAP模型组、毛果芸香碱组,每组6只。毛果芸香碱组小鼠腹腔给药毛果芸香碱(2.5 mg/kg体重)7 d,末次给药1 h后,APAP模型组和毛果芸香碱组腹腔注射APAP(400 mg/kg体重),空白对照组腹腔注射等量生理盐水,APAP注射后12 h处死所有小鼠,采集血液和肝脏等组织样本。血清生化检测仪检测血清肝功指标天门冬氨酸氨基转移酶(Aspartate aminotransferase, AST)、丙氨酸氨基转移酶(Alanine aminotransferase, ALT),采用HE染色方法检测肝脏病理评估毛果芸香碱对APAP诱导肝损伤的药效作用;Western blot方法检测肝脏增殖细胞核抗原(Proliferating cell nuclear antigen, PCNA)及细胞外调节蛋白激酶(Extracellular regulated protein kinases1/2, ERK1/2)蛋白表达,探索肝细胞增殖作用及其可能的机制。结果 与空白对照组比较,APAP模型组小鼠的ALT和AST含量均显著升高(P<0.001),表明APAP模型建立成功;与APAP模型组比较,毛果芸香碱组小鼠血清AST含量显著降低(P<0.01),ALT含量无统计学差异(P>0.05)。HE染色病理学结果显示,与空白对照组比较,APAP模型组小鼠肝脏出现肝小叶中心坏死、肝内出血和细胞核消失或固缩;与APAP模型组比较,毛果芸香碱组小鼠改善了肝小叶中心坏死、肝内出血和细胞核固缩。Western blot结果显示,与空白对照组和APAP模型组比较,毛果芸香碱组小鼠肝脏PCNA蛋白表达水平升高,差异有统计学意义(P<0.01);与空白对照组和APAP模型组比较,毛果芸香碱组小鼠p-ERK/ERK蛋白表达水平升高,差异有统计学意义(P<0.01)。结论 毛果芸香碱能减轻APAP所致肝损伤,其分子机制可能是毛果芸香碱通过激活肝细胞中m3AchR-ERK1/2信号通路进而促进肝细胞增殖来发挥作用。

硝酸毛果芸香碱滴眼液稳定性提升的处方工艺研究

目的 立足工业化生产,开发提高硝酸毛果芸香碱滴眼液稳定性的处方工艺。方法 通过阅读文献、剖析参比制剂特性,设计并筛选出合适的处方工艺。结果 采用低温配制的方法、控制处方pH值、筛选合适的滤膜等措施,得到可以工业化生产的处方工艺。该处方中硝酸毛果芸香碱为主要成分、硼酸和枸橼酸钠为pH调节剂、氯化钠为渗透压调节剂、苯扎氯铵为抑菌剂。结论 本研究设计的硝酸毛果芸香碱滴眼液处方工艺合理,稳定性较原研制剂优越,可以进行大规模的工艺化生产。

基于严重痫样发作行为的小鼠匹罗卡品颞叶癫痫模型构建

目的探讨C57BL/6J亚系小鼠经腹腔注射匹罗卡品建立颞叶癫痫模型的方法,总结可用于预测造模成功的癫痫发作急性期行为学表现,为癫痫研究提供可行的造模方案。方法选择30只C57BL/6J亚系小鼠(主要研究对象)和40只C57BL/6N亚系小鼠(对照),通过单次腹腔注射匹罗卡品的方法诱导小鼠癫痫发作从而建立颞叶癫痫模型。两种亚系的小鼠各分为3组,分别经腹腔注射300 mg/kg、330 mg/kg或360 mg/kg的匹罗卡品,观察并比较两种亚系小鼠的运动性癫痫发作的行为学表现,并在造模后第7天开始连续监测小鼠的自发性癫痫发作(spontaneous recurrent seizures,SRS)行为。造模后28 d,处死小鼠并观察其海马的病理改变情况。结果注射匹罗卡品后,C57BL/6N亚系小鼠表现出典型的运动性癫痫发作,而后进入癫痫持续状态(status epilepticus,SE);C57BL/6J小鼠较少观察到典型的运动性癫痫发作及后续的SE,而更多表现为单侧肢体抽搐后持续数秒至数十秒的全身颤抖,本研究将此行为学表现定义为严重痫样发作(severe seizure,SS)。腹腔注射330 mg/kg和360 mg/kg匹罗卡品后,在癫痫急性期出现过SS的C57BL/6J小鼠,经过潜伏期后可出现SRS,C57BL/6J亚系小鼠造模后SRS的比例(70%)和经历过SE后出现SRS的C57BL/6N亚系小鼠(75%)相近。造模后28 d,C57BL/6J小鼠海马出现颞叶癫痫的特征性病理改变,包括苔藓纤维出芽和神经元丢失。结论C57BL/6J亚系小鼠经腹腔注射匹罗卡品诱导癫痫模型时,造模成功的行为学标准可以是SE的发生,也可以是SS的出现。

去乙酰毛花苷联合美托洛尔对心力衰竭患者心功能及血清细胞因子水平的影响

目的探讨去乙酰毛花苷注射液联合美托洛尔对心力衰竭(简称心衰)患者心功能及其血清细胞因子水平的影响。方法回顾性选取70例心衰患者作为研究对象,根据治疗方案的不同,将患者分为对照组、联合用药组,每组各35例。对照组口服美托洛尔缓释片治疗,联合用药组采用去乙酰毛花苷注射液联合美托洛尔缓释片治疗。对比两组临床疗效、不良反应发生率,以及治疗前后心功能指标[左心室射血分数(LVEF)、心脏指数、每搏输出量]、血流动力学参数[舒张末期流速(end-diastolic flow velocity,EDFV)、收缩期峰值流速(peak systolic flow velocity,PSFV)、血氧饱和度(pulse oximetry,SpO_(2))、中心静脉压]和血清细胞因子[NT-proBNP、肌酸激酶同工酶(CK-MB)、血管生成素样蛋白2(angiopoietin-like protein 2,ANGPTL2)、血管生成素样蛋白7(ANGPTL7)]水平。结果联合用药组治疗总有效率、每搏输出量、心脏指数、LVEF、EDFV、PSFV、SpO_(2)水平显著高于对照组,血清NT-proBNP、CK-MB、ANGPTL2、ANGPTL7、中心静脉压均显著低于对照组(均P<0.05);两组不良反应总发生率间的差异无统计学意义(P>0.05)。结论利用去乙酰毛花苷注射液联合美托洛尔缓释片治疗心衰,可减轻机体炎症反应、降低心肌损伤程度、改善心功能,且不良反应少。

毛果芸香碱对映体的毛细管电泳手性分离

采用 β_环糊精及其衍生物作为手性选择剂对毛果芸香碱对映体进行了分离 ,研究了环糊精类型和浓度对分离的影响 ,同时考察了背景电解质 pH、操作电压和温度等因素对对映体分离的影响 ;结果表明 ,采用羟丙基_β_环糊精 (HP_β_CD)可以使毛果芸香碱对映体达到基线分离 ,优化条件下手性分离度可达2.79,为此类药物提供了一种简便、快速的毛细管电泳手性分离分析方法。

毛果芸香碱和布林佐胺滴眼液对兔眼表组织的影响

目的:探讨抗青光眼药物毛果芸香碱和布林佐胺滴眼液及各自的防腐剂三氯叔丁醇和氯化苯甲烃胺(BAC)对兔眼表组织的影响。方法:选取新西兰大白兔15只,分为正常对照组(3只)、毛果芸香碱组(A组,6只)和布林佐胺组(B组,6只)。用药组右眼使用青光眼药物毛果芸香碱或布林佐胺滴眼液,左眼使用对应的防腐剂三氯叔丁醇或BAC,连续用药30d。取球结膜组织行苏木素-伊红(HE)染色计数结膜上皮层炎症细胞数;角膜行扫描电镜检测并进行上皮损害分级评分。结果:与正常对照组比较,布林佐胺滴眼液(P<0.01)和防腐剂BAC(P<0.01)导致球结膜上皮层炎症细胞浸润增多;毛果芸香碱滴眼液(P>0.05)和三氯叔丁醇(P>0.05)引起的球结膜炎症细胞增多不明显。布林佐胺滴眼液导致球结膜炎症细胞增加的程度较毛果芸香碱滴眼液严重(P<0.01)。青光眼药物和防腐剂均可导致兔角膜上皮超微结构的损伤,表现为角膜上皮细胞六边形结构变为不规则、边界不清、细胞膜皱缩、细胞间距增宽、上皮细胞表面的微绒毛丢失、细胞空洞和暗细胞增加。布林佐胺滴眼液和BAC引起的角膜损害较毛果芸香碱滴眼液(P<0.01)和三氯叔丁醇明显(P<0.05)。各组左右两眼的球结膜炎症细胞数和角膜损害评分均无显著差异。结论:布林佐胺滴眼液和防腐剂BAC使用1个月后可导致兔球结膜炎症细胞增加和角膜上皮细胞损伤,毛果芸香碱滴眼液和三氯叔丁醇主要引起角膜上皮超微结构的变化;防腐剂是引起眼表损伤的主要原因。布林佐胺滴眼液对兔眼表的损伤较毛果芸香碱滴眼液严重。

匹罗卡品致大鼠癫痫持续状态后海马神经元凋亡的动态观察

目的探讨癫痫持续状态(status epilepticus,SE)后海马细胞凋亡的发生及其与caspase-3表达的关系。方法采用匹罗卡品诱发大鼠SE模型,用TUNEL染色和免疫组化技术检测海马神经元凋亡和caspase-3表达的动态变化。结果正常对照组大鼠海马未见TUNEL阳性细胞;SE后6 h,开始出现少量TUNEL阳性细胞;SE后72 h,达到高峰;SE后7 d,TUNEL阳性细胞开始减少。正常对照组大鼠海马可见少量caspase-3阳性表达;SE后6 h,大鼠海马caspase-3阳性表达增多,主要集中于CA1和CA3区;SE后48 h,caspase-3表达达到高峰;SE后72 h^7 d,caspase-3阳性染色细胞数开始减少;但仍显著多于对照组,差异有极显著意义(P<0.01)。结果显示caspase-3表达分布与TUNEL染色基本一致,caspase-3表达高峰早于TUNEL所示凋亡细胞出现的高峰。结论神经元凋亡参与了SE后海马神经元迟发性死亡过程并与caspase-3的激活有密切的关系。