纳洛酮对神经元GluR2表达的影响及缺氧损伤保护作用

目的:研究纳洛酮对缺氧损伤神经元膜表面AMPA(amino-3-hyoroxy-5-methylisoxazole-4-propionicacid,α-氨基-3-羟基-5甲基-4-异口恶唑丙酸)受体在谷氨酸受体第二亚单位(GluR2,glutamic acid receptor 2)表达的影响及其对神经细胞凋亡的保护作用。方法:取体外培养12 d的大鼠海马神经元,建立缺氧再灌注损伤模型,分别采用流式细胞和双重免疫荧光技术定量观察神经元凋亡数量、胞膜表面GluR2含量变化及纳洛酮的调节作用。结果:纳洛酮可上调缺氧损伤后神经元膜GluR2的含量(P<0.05),减少缺氧诱导的神经元凋亡的数量(P<0.01)。结论:纳洛酮通过上调神经元膜表面GluR2含量,减少神经元凋亡,减轻继发性脑损害。

淫羊藿素通过CXCR4/SDF-1信号通路促进小鼠成骨细胞成熟和矿化

目的:研究淫羊藿素是否通过CXC趋化因子受体4(CXCR4)/基质细胞衍生因子1(SDF-1)信号通路促进小鼠成骨细胞MC3T3-E1成熟和矿化。方法:将体外培养的MC3T3-E1细胞分为空白对照组、AMD3100组、淫羊藿素组、淫羊藿素加AMD3100组。药物处理24 h时,采用实时定量RT-PCR和蛋白质印迹法检测CXCR4、SDF-1和成骨相关基因和蛋白的表达;药物处理3、6 d时,采用碱性磷酸酶(ALP)试剂盒检测ALP的活性;药物处理14 d时,采用茜素红染色法检测钙化结节。结果:实时定量RT-PCR检测结果显示,与空白对照组比较,淫羊藿素组CXCR4、SDF-1、Runx2和OPG基因表达量增加(P<0.05或P<0.01),而加入CXCR4拮抗剂AMD3100后,CXCR4基因相对表达量减少(P<0.05)。蛋白质印迹法检测结果显示,与空白对照组比较,淫羊藿素组CXCR4、SDF-1、Runx2和OPG蛋白的相对表达量增加(均P<0.01),而加入AMD3100后,CXCR4、SDF-1、Runx2和OPG蛋白表达量减少(均P<0.01)。药物处理第3天和第6天时,淫羊藿素组ALP活性明显高于空白对照组(均P<0.01),而加入AMD3100后ALP活性显著降低(均P<0.01)。药物处理14 d时,与空白对照组比较,淫羊藿素组茜素红染色面积增加,而加入AMD3100后茜素红染色面积明显减少。结论:淫羊藿素可能通过CXCR4/SDF-1信号通路促进小鼠成骨细胞成熟及矿化。

半胱氨酰白三烯受体对小鼠小胶质细胞吞噬功能的调节作用

目的:研究半胱氨酰白三烯(CysLT)受体(CysLT_1R和CysLT_2R)对小鼠BV2小胶质细胞吞噬功能的调节作用。方法:以经典炎症激活剂脂多糖和CysLT受体激动剂LTD_4处理BV2细胞,采用免疫荧光计数法和流式细胞仪检测BV2细胞吞噬功能,免疫荧光共染法观察BV2细胞中CysLT_1R和CysLT_2R表达分布。结果:脂多糖和LTD_4均能增强BV2细胞吞噬功能,而CysLT_1受体选择性拮抗剂孟鲁司特和CysLT_2受体选择性拮抗剂HAMI 3379均能抑制脂多糖和LTD_4诱导的BV2细胞吞噬功能增强;脂多糖和LTD_4激活BV2细胞后可引起CysLT_1R和CysLT_2R在细胞内分布变化,两种亚型的受体分布变化趋势基本一致,且存在共表达。结论:CysLT_1R和CysLT_2R均可以调节BV2细胞的吞噬功能,且两者具有协同性。

胰岛素样生长因子-1对甲状腺相关眼病眼眶成纤维细胞的促生长作用

背景甲状腺相关眼病（TAO）是一种自身免疫性疾病，目前对其发病机制的研究主要集中在共同抗原上。胰岛素样生长因子-1（IGF．1）功能的发挥需要IGF-1受体（IGF-1R）的参与，IGF-1在促甲状腺激素受体（TSHR）的信号传导通路中也起着重要作用。目的探讨IGF-1对TAO患者眼眶成纤维细胞（OFs）增生及IGF-1R、TSHR表达的影响，探讨IGF-1在TAO发病机制中的作用。方法收集2016年3-6月于四川大学华西医院行TAO眼眶脂肪切除术取得的眶组织标本17例17份及正常人美容手术取得的眼眶组织4人4份，采用含体积分数5％胎牛血清的DMEM／F12培养液培养OFs。分别于培养液中加入不同质量浓度（O、50、100、125μg／L）的IGF-1，采用MTS比色法绘制OFs生长曲线，评估IGF-1对TAO和正常来源OFs增生的影响；采用流式细胞术测定不同质量浓度IGF-1对TAO和正常来源OFs中IGF-1R、TSHR表达水平的影响。结果TAO患者与正常人来源的OFs均生长良好，呈梭形，细胞质丰富，TAO患者与正常人来源的OFs形态学特点一致。培养的OFs波形蛋白呈阳性反应，结蛋白、S-100、肌红蛋白和角蛋白均呈阴性反应。不同质量浓度IGF-1作用后TAO组和正常组OFs增生均呈逐渐上升趋势，总体比较差异有统计学意义（F分组=219．639，P〈0．001；F质量浓度=17．752，P〈0．001），以100μg／LIGF-1的促进作用最强。0、50、100、125μg／LIGF-1作用后TAO组OFs中IGF-1R表达量分别为（0．0091±0．0087）％、（0．0953±0．0233）％、（0．0837±0．0227）％和（0．0709±0．0241）％，正常组OFs中IGF-1R表达量分别为（0．0023±0．0006）％、（0．0093±0．0012）％、（0．0073±0．0015）％和（0．0083±0．0012）％，其中50、100和125μg／LIGF-1作用后各组OFs中IGF-1R表达量均高于无添加IGF-1者，TAO组OFs细胞中IGF-1R的表达量均明显高于正常组，差异均有统计学意义（均P〈0．05）；0、50、100和125μg／LIGF-1作用后TAO组和正常组OFs中TSHR表达量的总体比较差异均无统计学意义（F分组=0．133，P〉0．05；F质量浓度=0．004，P〉0．05）。结论IGF-1对TAO患者和正常人OFs的生长均有促进作用并能上调OFs中IGF-1R的表达，但其对OFs中TSHR的表达变化无明显影响。

Etiological characteristics and treatment of tardive dyskinesia

OBJECTIVE ： The pathogenesis of tardive dyskinesia （TD） is complicated and uncertain, Thus, there is not any effective treatment for it. The psychiatrists pay more and more attention to TD, which lasts for a long time and is difficult to treat. DATA SOURCES： A computer-based online search of Medline database was undertaken to identify articles about the feature of etiology and the progression of treatment for TD published in English by using the keywords of ＂rD, etiology, pathogenesis＂ and ＂TD, therapy, drug treatment＂. Meanwhile, Chinese articles about the feature of etiology and the progression of treatment for TD were searched in Wanfang database and China journal full-text database, and the keywords were ＂TD, etiology, pathogenesis＂ and ＂TD, therapy, drug Treatment＂ in Chinese. STUDY SELECTION： Articles met the following inclusion criteria were selected in this paper. Inclusion criteria： （1） Researches of randomized blind control design, before and after control design and retrospective. （2） Researches of the feature of etiology and the progression of treatment for TD. Exclusion criteria： the repetitive researches and individual reports. DATA EXTRACTION ： Totally 65 articles related the feature of etiology and the progression of treatment for TD of randomized blind control design, before and after control design and retrospective studies were collected, and 53 of them were accorded with the inclusion criteria. Of the 12 excluded ones, 8 were concerning with genetics, 4 were repetitive researches. DATA SYNTHESIS ： The feature of etiology for TD includes：（1） Hypothesis of dopamine receptor super-sensitivity： The dopamine receptor is persistently blocked, so it will result in functional disorder in CNS, and then TD may take place. （2）） Hypothesis of neuronal degeneration： The concentration of aminosuccinic acid and glutamic acid will increase after the antipsychotic used for a long time and this will result in neuronal degeneration through glutamic acid receptor in the postsynaptic membrane; meanwhile with free radical, the nerve cells of corpus striatum may degenerate and become necrosis. （3） Sex and age： The females and gerontal patients are liability to the TD disease. It is may related to the lower estrogen. （4） Molecule heredity： TD may association with the dopamine and 5-HT receptor gene polymorphism. （5） Other theories： Hypofunction of γ-amino-butyri acid （GABA）, hypothesis of noradrenaline 5-serotonin and nutrition metabolism can cause TD disease. Treatlent for TD： （1） Dopamine receptor agonist： The therapeutic effect is not satisfactory, especially for gerontism females. （2） Oxygen free radical scavenger： As represent of vitamin E, it can clear out free radicals and reduce the potential cytotoxic effect of free radicals. （3） Calcium channel blocker： This maybe related to block calcium ions releasing from muscle cells and inhibit muscle convulsion; therefore, it can be used for symptomatic treatment. （4） GABA receptor agonist： It is more effective for the prominent dysmyotonia than dancing slowly symptom. （5） Antipsychotic： There is some therapeutic effect with ciozapine, but the effect will reduce because of the age growing up and the symptom exacerbating. （6） Other therapies： Valproate sodium, cyproheptadine, melatonin, branched chain amino acid, ahalysantinfarctasum, electric acupuncture and injection ad acumen, traditional Chinese drug have a certain effects on TD. Prevention of TD： The serum creatine phosphokinase （CPK） combined with symptoms should be checked regularly so as to early discovery TD. CONCLUSION ： （1） Etiology of TD： The hypothesis of dopamine receptor super-sensitivity is denyed; the hypothesis of neuronal degeneration is approved in academic circles; the sex and age is a finding of generally received; but the dopamine and 5-HT receptor gene polymorphism, hypofunction of GABA, noradrenaline, 5-serotonin and nutrition metabolism cannot explain the pathogenesis of TD. （2） Treatment for TD： The therapeutic effect of dopamine receptor agonist is not satisfactory; the oxygen free radical scavenger maybe effective; calcium channel blocker maybe used for symptomatic treatment; GABA receptor agonist maybe more effective for the prominent dysmyotonia than dancing slowly symptom; the consequence of antipsychotic is discrepancy; other therapies maybe use to adjunctive therapies. （3） As far as prevention of TD is concerned, and the serum CPK combined with symptoms should be checked regularly so as to early discovery TD.

大麻类物质体内代谢毒理及检测研究进展

大麻是国际管制的麻醉药品,因其价廉且摄取方便而长期在世界范围内被广泛使用。近年来,作为大麻的替代品,各种新型合成大麻素以"香料"的形式不断涌现并被非法使用。摄入大麻类物质后,使用者的短期记忆、感觉敏感性以及运动神经协调性等都会受到损伤,因此,生物检材中大麻类物质的检测及结果评判十分重要。本文主要围绕大麻及合成大麻素等大麻类物质的药理毒理特点、体内代谢、原体及代谢产物的检测方法等方面的研究进行综述。