吗啡依赖及戒断后大鼠垂体远侧部促肾上腺皮质激素细胞和甲状腺刺激激素细胞组织学研究

目的：观察吗啡依赖大鼠及戒断大鼠垂体远侧部促肾上腺皮质激素（ＡＣＴＨ）细胞和甲状腺刺激激素（ＴＳＨ）细胞的变化。方法：皮下规律注射吗啡建立大鼠吗啡依赖模型,戒断后造成戒断模型并用免疫组织化学和图像分析方法检测吗啡依赖及戒断后大鼠垂体远侧部ＡＣＴＨ细胞和ＴＳＨ细胞的变化。结果：吗啡依赖大鼠ＡＣＴＨ细胞和ＴＳＨ细胞免疫反应减弱（Ｐ＜０．０１）,短期戒断后这些改变仍持续存在。结论：吗啡可导致大鼠脑垂体分泌功能紊乱,且短期戒断后未能完全恢复。     

吗啡依赖及乌拉地尔戒断大鼠垂体远侧部 ACTH 细胞免疫组织化学研究

目的观察吗啡依赖及乌拉地尔戒断大鼠垂体远侧部促肾上腺皮质激素(ACTH)细胞的变化。为探讨乌拉地尔对吗啡依赖大鼠 ACTH 细胞分泌功能影响提供形态学依据。方法用皮下注射吗啡法建立雄性大鼠吗啡依赖模型，戒断组给吗啡依赖大鼠侧脑室注射(IVC)乌拉地尔。用免疫组织化学和图像分析方法观察吗啡依赖大鼠及乌拉地尔戒断大鼠垂体远侧部 ACTH 细胞的变化。结果吗啡依赖大鼠及乌拉地尔戒断大鼠 ACTH 细胞免疫反应减弱(P＜0．01)。侧脑室注射乌拉地尔正常大鼠 ACTH 细胞免疫反应增强(P＜0．05)。结论乌拉地尔可引起吗啡依赖大鼠垂体远侧部 ACTH 细胞分泌功能减弱，引起正常大鼠 ACTH 细胞分泌功能增强。     

吗啡依赖及乌拉地尔戒断大鼠垂体远侧部TSH细胞免疫组织化学研究

用皮下注射吗啡 ,建立雄性大鼠吗啡依赖模型。戒断组给吗啡依赖大鼠侧脑室注射乌拉地尔( IVC)。阳性对照组给正常大鼠 IVC,阴性对照组给正常大鼠 IVC相同体积生理盐水。用免疫组织化学方法观察吗啡依赖大鼠 ,IVC戒断大鼠和 IVC正常大鼠垂体远侧部 TSH细胞的变化。结果发现 ,吗啡依赖大鼠和IVC戒断大鼠 TSH免疫反应细胞的面数密度减少 ,免疫反应减弱 ( P <0 .0 1)。阳性对照组大鼠 TSH免疫反应细胞的面数密度增大 ,免疫反应增强 ( P <0 .0 5 )。本文对吗啡及乌拉地尔所引起大鼠垂体远侧部 TSH免疫反应细胞变化机理进行了探讨     

海洛因依赖对大鼠垂体促肾上腺皮质激素阳性细胞和血清皮质醇的影响

目的探讨海洛因依赖对大鼠垂体远侧部促肾上腺皮质激素细胞表达促肾上腺皮质激素(ACTH)的影响以及血清皮质醇(COR)的改变,并探讨引起变化的可能机制。方法成年雄性SD大鼠55只,随机分为正常对照组、盐水对照组及海洛因依赖组。皮下注射海洛因,建立海洛因依赖大鼠模型,分别于模型建立的第10、17、24、31、38天取垂体组织。应用免疫组织化学SABC法、图像分析法及放射免疫方法进行研究。结果与正常对照组和盐水对照组比较,海洛因依赖组大鼠垂体远侧部ACTH阳性细胞免疫反应明显减弱,与正常及盐水对照组比较有显著性差异;图像分析显示,海洛因依赖组ACTH阳性细胞的平均灰度值增高(P0.05〉。放射免疫法测定结果显示,海洛因依赖组血清COR含量较正常对照组明显降低,经统计处理,差异有统计学意义(P0.05)。结论在海洛因依赖期间,垂体远侧部ACTH阳性细胞表达减少,血清COR含量降低,提示在大鼠海洛因依赖期间,垂体-肾上腺轴功能受到抑制。     

颅脑损伤大鼠垂体促肾上腺皮质激素细胞免疫组织化学变化

目的: 观察颅脑损伤大鼠垂体远侧部促肾上腺皮质激素(ACTH)细胞的免疫组织化学变化。方法: 采用落体法致大鼠颅脑损伤; 伤后24 h断头处死,用免疫组织化学和图像分析方法观察颅脑损伤后大鼠ACTH细胞的变化。结果: 颅脑损伤大鼠ACTH细胞的面数密度、平均吸光度及免疫组化反应均显著强于对照组(P<0.05)。结论: 颅脑损伤大鼠ACTH细胞合成分泌促肾上腺皮质激素功能增强。     

吗啡依赖及乌拉地尔戒断大鼠垂体远侧部ACTH细胞免疫组织化学 …

目的：观察吗啡依赖及乌拉地尔戒断大鼠垂体远侧部促肾上腺皮质激素（ＡＣＴＨ）细胞的变化。为探讨乌拉地尔及吗啡依赖大鼠ＡＣＴＨ细胞分泌功能影响提供形态学依据。方法：用皮下注射吗啡法建立雄性大鼠吗啡依赖模型,戒断组给吗啡依赖大鼠侧及室注射（ＩＶＣ）乌拉地尔,用免疫组织化学和图像分析方法观察吗啡信赖大鼠及乌拉地尔戒断大鼠垂体远侧部ＡＣＴＨ细胞的变化。结果：吗啡依赖大鼠及乌拉地尔戒断大鼠ＡＣＴＨ细胞免疫反应     

促肾上腺皮质激素应用于原发性肾病综合征频复发及激素依赖患儿疗效及安全性初探

目的探讨促肾上腺皮质激素（ACTH）治疗原发性肾病综合征（PNS）频复发或激素依赖患儿的疗效和安全性。方法以符合PNS频复发或激素依赖患儿为ACTH组,每月给予ACTH缓慢静滴3—5d,并逐渐激素减量至停药;以激素维持治疗PNS患儿为对照组。观察治疗前与治疗后3、6和12个月两组激素剂量、复发次数、肾上腺皮质功能及不良反应。结果2010年9～12月ACTH组纳入14例,对照组纳入6例。（1）ACTH组12／14例均能顺利将激素减量至停药,无复发;2例因感染尿蛋白波动,感染控制后尿蛋白仍阳性,激素加量至尿蛋白转阴后减量,继续ACTH冲击,其中1例因感染后再次复发,停用ACTH改用他克莫司。ACTH组13例进入分析。（2）ACTH组治疗后3、6和12个月激素剂量与治疗前差异有统计学意义（P〈0．05）;对照组治疗后3、6和12个月激素剂量与治疗前差异无统计学意义（P〉0．05）;ACTH组与对照组治疗后6、12个月激素剂量差异有统计学意义（P〈0．05）。③治疗前ACTH组和对照组均存在肾上腺皮质功能低下,ACTH组13例在治疗3、6和12个月分别有9、11和13例肾上腺皮质功能恢复正常;对照组6例12个月治疗期间均表现为肾上腺皮质功能低下。（4）ACTH组观察到腰部皮疹和心率加快各1例。（5）ACTH组治疗后6、12个月身高与治疗前差异有统计学意义,对照组治疗前后身高差异无统计学意义（P〉0．05）。ACTH组治疗前后体重差异无统计学意义,对照组治疗后6、12个月体重与治疗前差异有统计学意义（P〈0．05）。（6）ACTH组和对照组治疗前后骨密度差异均无统计学意义（P〉0．05）。结论ACTH治疗PNS频复发或激素依赖患儿有一定疗效,可避免长期激素治疗的不良反应,值得进一步探讨。     

老龄大鼠下丘脑及垂体促肾上腺皮质激素样和β－内啡肽样免疫阳性细胞的相关变化

正常Ｗｉｓｔａｒ大鼠，雌雄不拘，老龄鼠（２０个月）８只，壮龄鼠（８个月）１３只。石蜡切片，ＡＢＣ法染色，ＤＡＢ显色．光镜下观察下丘脑及垂体促肾上腺皮质激素样免疫反应（ＡＣＴＨ－ｉｒ）和β－内啡肽样免疫反应（β－ＥＮｒｙ－ｉｒ）阳性细胞的形态、分布和计数。图像分析仪测其灰度值、结果：下丘脑弓状核β－ＥＮＤ－ｉｒ阳性细胞可分为胞核不明显、着色较深、数量较多的强阳性细胞和胞核明显、着色较淡、数量较少的弱阳性细胞；ＡＣＴＨ－ｉｒ阳性细胞，胞核明显，胞体较大，类似于β－ＥＮＤ－ｉｒ弱阳性细胞。老龄大鼠这两种阳性细胞出现率均明显低于壮龄大鼠。弓状核和垂体前叶ＡＣＴＨ－ｉｒ阳性细胞灰度值老龄鼠显著低于壮龄鼠，而β－ＥＮＤ－ｉｒ阳性细胞灰度值老龄鼠则显著高于壮龄鼠。     

慢性吗啡处理及戒断后大鼠前额皮质、海马和杏仁核中Parvalbumin的表达变化

为了观察慢性吗啡处理及戒断后大鼠前额皮质、海马和杏仁核中parvalbumin(PV)的表达变化,为其功能的研究提供形态学依据,本实验将30只雄性SD大鼠随机分为吗啡依赖组和生理盐水对照组。吗啡依赖组大鼠腹膜腔注射吗啡,2次/d,起始剂量为5mg/kg,逐日递增5mg,至第10d为50mg/kg;对照组注射同体积的生理盐水。于末次注射后动物分别存活3h、3d和14d。用免疫组化方法和相对平均灰度值检测前额皮质、海马和杏仁核内PV的表达。结果显示:在生理盐水处理组各存活时间点,前额皮质、海马和杏仁核内PV的表达相同。和生理盐水对照组相比,3h时海马和杏仁核内PV的表达明显增加(P<0.05),但前额皮质内PV的表达减少。第3d时,海马CA1、CA3、CA4、齿状回和杏仁核内PV的表达减少,但CA2区PV表达继续增加,而前额皮质的表达开始恢复。至第14d时,CA2区PV的表达开始恢复,但CA1、CA4和杏仁核内PV的表达又开始增加,明显高于第3d组(P<0.05)。以上结果提示慢性吗啡处理及戒断后PV的表达具有区域特异性和时相特异性;这种变化在戒断早期可能主要与躯体依赖相关,而戒断晚期主要与精神依赖相关。     

大鼠胃肠道促肾上腺皮质激素免疫反应性细胞的观察

用ＰＡＰ免疫组织化学方法观察了１０只正常大鼠胃肠道促肾上腺皮质激素免疫反应性细胞的分布及形态特点。结果如下：促肾上腺皮质激素免疫反应性细胞主要分布在胃窦、胃体及十二指肠。细胞形态多样，有柱状、锥体状及特殊的杯状等。促肾上腺皮质激素免疫反应性细胞基部紧贴基膜，免疫反应性强弱不一。多数细胞促肾上腺皮质激素免疫反应性阳性物质可达肠倥面或腺腔面，有时腔内可见到有促肾上腺皮质激素免疫反应性阳性物质，说明促肾     

Height-dependent difference in the expression of naloxone-induced withdrawal jumping behavior in morphine dependent rats

Opiate withdrawal syndrome may motivate opiate seeking and taking. Thus, development of an effective medical treatment for these symptoms is a primary research goal and strongly relies on improved experimental models. Opiate withdrawal syndrome is characterized by several behavioral signs such as wet dog shake, teeth chattering, sniffing, scratching, chewing, diarrhea, rearing, ptosis and jumping. The goal of present study was to evaluate the impact of the cylindrical chamber height on the expression of jumping behavior in morphine dependent rats. Adult male Wistar rats were rendered dependent on morphine by subcutaneous (s.c.) injection of morphine sulfate (10 mg/kg) with an interval of 12 h for 9 days. On day 10, 2 h after morphine injection, rats were injected with naloxone (1 mg/kg, i.p.). Naloxone-induced jumping was monitored during a period of 30 min in a clear cylindrical Plexiglas test chamber with the floor covered by woodchip. The chambers had the same diameter (35 cm), but the heights of chambers were different (30, 40, 50, 60, 70 and 80 cm). Incidence and frequency of jumping decreased with increasing the height of the test chambers (P<0.05). Altogether, these findings highlight the possibility of detecting height-dependent difference in the expression of naloxone-induced jumping behavior in morphine dependent rats.     

八肽胆囊收缩素对吗啡戒断大鼠蓝斑和中脑导水管周围灰质CREB及p-CREB的影响

目的 探讨八肽胆囊收缩素（CCK-8）及其受体拮抗剂对吗啡戒断症状的影响及其相应的信号转导机制。方法 建立吗啡依赖及戒断动物模型,每组6只。应用改良柳田知司法对戒断症状评分,并采用免疫组织化学方法检测大鼠蓝斑和中脑导水管周围灰质内CREB及p-CREB的变化。结果 腹腔及侧脑室注射CCK-8,CCK1及CCK2受体拮抗剂均可明显降低戒断症状评分;吗啡依赖后蓝斑和中脑导水管内p-CREB明显增高,戒断后蓝斑内p-CREB水平较依赖组进一步增高,而中脑导水管内p-CREB却较依赖组下降。腹腔及侧脑室注射CCK-8,CCK1及CCK2受体拮抗剂均可逆转戒断后中脑导水管内p-CREB的降低;腹腔及侧脑室注射CCK1 及CCK2受体拮抗剂可逆转戒断后蓝斑内p-CREB的升高,而CCK-8却对蓝斑内p-CREB的表达无明显影响。结论 CCK-8可通过对蓝斑和中脑导水管内p-CREB的调节减轻吗啡戒断症状,且表现出明显的脑区特异性。     

Extremely low-frequency electromagnetic field exposure during chronic morphine treatment strengthens downregulation of dopamine D2 receptors in rat dorsal hippocampus after morphine withdrawal

The aim of this study was to investigate the effect of extremely low-frequency electromagnetic field (ELF-EMF) exposure during morphine treatment on dopamine D2 receptor (D2R) density in the rat dorsal hippocampus following withdrawal. Rats were exposed to ELF-EMF (20 Hz, 14 mT) or sham exposed for 1h per day before injection of morphine (10mg/kg, i.p.) once daily for 12 days. The saline control group was sham exposed for the same period. Immunohistochemistry was used to detect the density of D2Rs on the 1st, 3rd and 5th morphine withdrawal days. The results showed that the density of D2Rs in sham-exposed morphine-treated rats on the 1st and 3rd days of morphine withdrawal was significantly lower than that of the saline control group. The ELF-EMF-exposed morphine group also exhibited a significantly lower density of D2Rs on the 1st and 3rd withdrawal days relative to the sham-exposed morphine group. However, the D2R density in both groups tended to recover as morphine withdrawal days increased. The results suggest that dorsal hippocampal D2Rs are sensitive to morphine withdrawal and that this is potentiated by ELF-EMF pre-exposure during morphine treatment.     

吗啡依赖与戒断大鼠伏隔核单胺、类阿片肽及下丘脑POMCmRNA改变的研究

研究了大鼠吗啡依赖与戒断状态和伏隔核类阿片肽、单胺、下丘脑ＰＯＭＣ基因表达改变之间的关系。发现在吗啡依赖状态伏隔核内β－内啡肽、脑啡肽、单胺释放以及下丘脑ＰＯＭＣ基因表达受抑制。戒断４８小时后强啡肽、ＮＥ、ＭＨＰＧ、５－ＨＴ升达高峰，后虽水平有所降低，但戒断１周时仍高于对照显著。深受抑制的ＤＡ、β－内啡肽释放以及ＰＯＭＣ基因表达在戒断后有所回升，但于戒断１周时还是显著低于对照水平。提出这些生化改变尤其是自从依赖形成后ＤＡ的持续低水平是吗啡依赖的机理之一。     

CCK-8及其受体拮抗剂对吗啡戒断大鼠额叶皮质及海马CREB与pCREB表达的影响

 目的：观察八肽胆囊收缩素（CCK-8）及其受体拮抗剂对吗啡戒断大鼠额叶皮质和海马cAMP反应元件结合蛋白（CREB）表达及其磷酸化（pCREB）的影响,初步探讨CCK-8调节吗啡戒断大鼠的受体后机制。方法：建立大鼠吗啡慢性依赖及纳络酮催促戒断模型,并给予CCK-8、CCK1受体拮抗剂L-364718和CCK2受体拮抗剂LY-288513慢性干预,应用Western blotting和免疫组织化学技术观察额叶皮质和海马CREB与pCREB 表达的变化。结果：（1） 正常组大鼠额叶皮质神经元胞浆、胞核均表达CREB蛋白,pCREB蛋白则仅在胞核中高表达;海马CA1区锥体细胞层神经元中,CREB蛋白在胞浆中高表达,胞核低表达,pCREB蛋白则仅在胞核中表达。（2） 慢性吗啡作用后CREB无明显变化,pCREB增加;急性纳洛酮催促戒断后CREB仍无明显变化,pCREB进一步升高。（3） 与戒断组相比,CCK-8、L-364718和LY-288513慢性干预对吗啡依赖戒断大鼠额叶皮质CREB蛋白表达无明显影响,pCREB蛋白表达均明显降低;L-364718和LY-288513慢性干预后,海马CREB与pCREB表达均明显降低,而CCK-8慢性干预对CREB蛋白表达无明显影响,仅pCREB蛋白表达明显降低。结论：CCK-8及其受体拮抗剂可能通过调节核转录因子CREB减轻吗啡戒断症状,并具有脑区特异性。     

Immunohistochemical localization of afatinib in male rat intestines and skin after its oral administration

Afatinib, a second-generation tyrosine kinase inhibitor, was designed to bind covalently to and irreversibly inhibit active ErbB family receptors. The major metabolites of afatinib in human plasma are adducts of afatinib covalently bound to plasma proteins via. the Michael addition reaction. These findings suggest that afatinib may form covalent bonds with proteins in tissue and be localized in tissue. However, there is no method for the specific detection of afatinib-protein conjugates localized in tissue. In this paper, we aimed to develop an immunohistochemical protocol to detect afatinib-protein conjugates. Immunostainings were performed with male rat intestinal tract and skin at 24 h after an oral administration of afatinib. In the intestinal tract, strong staining was observed in the ileum and colon, but only slight staining was observed in the duodenum and jejunum. In the skin, strong staining was observed in the epidermis, sebaceous glands and hair follicles. Immunohistochemistry for afatinib-protein conjugates could be a useful tool to detect the localization of such conjugates. This study is the first to elucidate the localization of afatinib-protein conjugates in the rat intestinal tract and skin and is expected to be of great use in efforts to clarify the mechanism underlying afatinib-induced diarrhoea or skin toxicities.     

Immunohistochemical localization of orexin/hypocretin-like immunoreactive peptides and melanin-concentrating hormone in the brain and pituitary of medaka

Orexin/hypocretin is a neuropeptide that is involved in the regulation of feeding behavior and the sleep–wakefulness cycle in mammals. Melanin-concentrating hormone (MCH) is believed to be another candidate involved in food intake in teleost fish as well. Thus, it is interesting to examine whether neural connections exist between the neurons producing these two hormones. We first examined the localization of orexin-like immunoreactivity (orexin-LI) in the brain of the medaka Oryzias latipes by using immunohistochemistry. We further examined the interaction between the orexin and MCH neurons in the medaka brain by performing double-staining immunohistochemistry. Orexin-LI cell bodies were located in the nucleus posterioris periventricularis (NPPv) of the hypothalamus, and orexin-LI fibers were detected not only in the hypothalamus but also extensively throughout the brain. Some orexin-LI fibers were in close contact with the MCH-immunoreactive (ir) cell bodies in the hypothalamus, as revealed by double-staining immunohistochemistry. Moreover, a few MCH-ir fibers were in close contact with the orexin-LI cell bodies. These results suggest that in the medaka brain, orexin performs various functions, including neuromodulation, and that neural connections exist between the orexin and MCH neurons.