Lateral ventricle injection of orexin-A ameliorates central precocious puberty in rat via inhibiting the expression of MEG3

Background: Central precocious puberty (CPP) is characterized as increasing gonadotropin-releasing hormone (GnRH) release. Orexin-A has also been shown to affect GnRH release. However, there are few reports about the effect of orexin A on the treatment of CPP. Methods: After establishing the precocious puberty model, the rats were divided into four groups: normal control, precocious puberty rats, precocious puberty rats treated with normal saline and precocious puberty rats treated with orexin-A. The vaginal opening time, second estrus cycle, ovarian index and uterus index of rats in each group were detected. qRT-PCR was performed to examine the expression of MEG3 and kisspeptin in rats. HT22 cells were transfected with pcDNA-MEG3 to detect the expression of Kisspeptin. Results: In this study, we found that orexin-A not only delayed the day of vaginal opening and regular estrus cycle days but also decreased the ovarian index and uterus index in rats with CPP. In addition, orexin-A reversed the up-regulation of MEG3 and kisspeptin in rats with CPP. In HT22 cells, the mRNA and protein level of kisspeptin were enhanced by pcDNA-MEG3. Conclusion: Our results suggest that orexin-A ameliorates central precocious puberty in rat and MEG3 might be involved in this effect, suggesting that MEG3 might be a novel target in treating central precocious puberty.     

刺五加脑灵液联合扎来普隆治疗失眠症的临床研究

目的探讨刺五加脑灵液联合扎来普隆片治疗失眠症的临床疗效。方法选择2018年2月-2019年2月在成都市第四人民医院诊治的失眠症患者86例,根据用药的差别分为对照组(43例)和治疗组(43例)。对照组睡前口服扎来普隆片,5 mg/次,1次/d;治疗组在对照组的基础上口服刺五加脑灵液,10 mL/次,2次/d。两组患者均经10 d治疗。观察两组患者临床疗效,同时比较治疗前后两组患者PSQI、AIS、PHQ-9、WEMWBS和QLQ-C30评分,睡眠质量以及血清血清5-羟色胺(5-HT)、脑源性神经营养因子(BDNF)、白细胞介素-1β(IL-1β)、食欲素A(Orexin-A)、肿瘤坏死因子-α(TNF-α)和IL-2水平。结果治疗后,对照组临床有效率为81.40%,显著低于治疗组的97.67%,两组比较差异有统计学意义(P<0.05)。经治疗,两组患者PSQI评分、AIS评分和PHQ-9评分均显著降低(P<0.05),但WEMWBS评分和QLQ-C30评分均显著升高(P<0.05),且治疗组这些评分项目改善更明显(P<0.05)。经治疗,两组患者总睡眠时间、REM睡眠时间、睡眠效率均提高(P<0.05),而睡眠潜伏期、觉醒时间均显著缩短(P<0.05),且以治疗组改善最为明显(P<0.05)。经治疗,两组患者血清5-HT、BDNF水平均明显升高(P<0.05),但IL-1β、Orexin-A、TNF-α、IL-2水平均明显降低(P<0.05),且以治疗组改善最为明显(P<0.05)。结论刺五加脑灵液联合扎来普隆片治疗失眠症可显著改善睡眠情况,提高生活质量,有利于焦虑抑郁状态和心理健康状态改善。     

Orexin-A对戊四氮慢性点燃癫痫大鼠学习记忆功能的影响

目的 探讨Orexin-A对戊四氮(PTZ)慢性点燃癫痫大鼠学习记忆功能的影响.方法 健康成年雄性Wistar大鼠随机分为对照组(生理盐水)和戊四氮组.点燃成功后每组分别脑室内注射10μlorexin-A(1.4 nmol/μl)或等量生理盐水,利用Morris水迷宫实验进行学习记忆能力检测.结果 致痫组戊四氮点燃癫痫的成功率达80%.与非致病组比较,慢性点燃大鼠搜寻平台的潜伏期明显延长(F=200.956,P<0.01);大鼠在平台所在象限的游泳时间和120 s内穿越平台区域的次数显著减少(P<0.01).脑室内注射orexin-A后,航行定位实验中致痫组大鼠逃避潜伏期明显缩短(P<0.01),非致痫组大鼠的逃避潜伏期也明显缩短(P<0.05).在空间搜寻实验中,orexin-A能明显增加穿越平台象限的次数[(10.83±1.84),(4.67±3.34)]和平台所在象限的游泳时间[(39.73±2.03)s,(33.76±2.96)s],尤其是对致痫组大鼠的怍用更明显(P<0.01),搜寻站台的效率明显提高.结论 Orexin-A能改善慢性点燃癫痫大鼠的空间学习记忆能力.

Abstract：

Objective To investigate the effects of orexin-A on learning and memory of pentyleneterazol (PTZ)-kindled epileptic rats.Methods Adult Wistar rats were randomly divided into control group(normal saline,NS)and PTZ group.The PTZ-kindled rats were randomly divided into,orexin-A group and NS group administrated by intracerebroventricular(i.c.v.)injection of 10μl orexin-A(1.4 nmol/μl)or 10μl NS.Using Morris water msze experiment,the ability of learning and memory was measured in all rats.Results Eighty percent of rats in PIZ group were kindled successfully after intraperitoneal injection of 35mg/kg PTZ every day for 30 days.Compared to control group,the escape latency in the PTZ-kindled rats was significantly increased in place navigation test(PNT)(F=200.956,P<0.01),whereas a remarkable reduction of time spent in the target quadrant and number of pool circlings in 120 seconds Was observed during probe trials.Following injection of orexin-A,the latency of escape platform was significantly declined in both PTZ-kindled((39.73±2.03)8,(33.76±2.96)s)and NS rats,increased the number of crossing the platform(10.83±1.80)vs(4.67±3.34).In addition,the treatment with orexin-A markedly increased swim velocity and number of pool circlings in beth groups(P<0.01),particularly to the PTZ-kindled rats.Conclusion Spatial learning and memory in the PIZ-kindled rats can be improved by treatment with orexin-A.     

睡眠呼吸暂停综合征儿童手术前后血浆食欲素A及感觉统合能力的变化

目的探讨睡眠呼吸暂停综合征（obstructivesleepapnea—hypopneasyndrome,OSAHS）儿童手术治疗前后血浆食欲素A（Orexin—A）水平变化及与感觉统合能力的关系。方法接受扁桃体切除和／或腺样体刮除术的儿童OSAHS患者120例,正常对照组30例。并按呼吸暂停低通气指数（apneahypopneain—dex,AHI）将OSAHS组分为轻度组（5次／h≤AHI〈20次／h,n=13）;中度组（20次／h≤AHI〈40次／h,n=76）;重度组（AHII〉40次／h,n=31）。同时按感觉统合能力评分还将OSAHS组分为正常组30名、轻度失调组47名、中度失调组28名、重度失调组15名。术前及术后6个月进行血浆Orexin—A水平测定及儿童感觉统合能力评定。结果OSAHS组患儿血浆Orexin—A浓度[（0．41±0．06）μ/ml]高于正常对照组[（0．31±0．04）μg／ml],差异有统计学意义CP〈0．01）;不同严重程度OSAHS患儿手术前后血浆Orexin—A浓度比较,轻度组[术前（0．33±0．02）μg／ml,术后（0．28±0．03）μ/ml]、中度组[术前（0．39±0．04）μ/ml,术后（0．29±0．03）μ/ml]、重度组[术前（0．49±0．04）μ/ml,术后（0．32±0．02）μg／ml]差异均有统计学意义（P〈0．01）。AHI指数与术前血浆Orexin—A浓度显著相关（r=0．803,P〈0．01）。感觉统合测评分值与术前血浆Orexin—A浓度也存在显著相关（r=-0．812,P〈0．01）。结论血浆Orexin—A浓度与儿童OSAHS严重程度及其行为认知能力改变关系密切,可能成为儿童OSAHS诊断和预后的血浆标志物。     

Biphasic effects of orexin-A on autonomic nerve activity and lipolysis

Previously, we showed that orexin-A, a 33-aa peptide, influences renal sympathetic nerve activity. Because the autonomic nervous system plays an important role in the regulation of lipid metabolism, we investigated the in vivo effects of orexin-A on the sympathetic nerve activity innervating white adipose tissue (WAT-SNA) and lipolysis. We found that intracerebroventricular (icv) administration of orexin-A at doses of 1mug/rat and 10ng/rat elevated and suppressed WAT-SNA, respectively. The effect of the high dose of orexin-A (1mug/rat) was eliminated by pretreatment with diphenhydramine hydrochloride, a histamine H(1) receptor antagonist. In contrast, the effect of the low dose of orexin-A (10ng/rat) was suppressed by thioperamide maleate salt, a histamine H(3) receptor antagonist. Moreover, icv administration of 1mug/rat and 10ng/rat of orexin-A increased and decreased the levels of plasma free fatty acids (FFAs), respectively. The effect of 1mug/rat of orexin-A on plasma FFA was eliminated by propranolol hydrochloride, a beta-adrenergic receptor blocker, and also by diphenhydramine. The effect of orexin-A at dose of 10ng/rat disappeared by pretreatment with atropine sulfate, a muscarinic receptor blocker, and thioperamide maleate salt. Our results suggest that high doses of orexin-A may regulate the lipolytic processes in adipose tissue through facilitation of the sympathetic nervous system, which is driven by histamine neurons through the H(1) receptor, and that the beta(3)-receptor may be involved in this enhanced lipolytic response. Low doses of orexin-A, on the other hand, may lower lipolysis by suppressing sympathetic nerve activity via the H(3)-receptor, and the muscarinic receptor may be related to this response.     

摩腹手法对睡眠剥夺大鼠脑内神经递质及Orexin-A的影响

[目的]观察和探讨摩腹手法对睡眠剥夺大鼠脑内神经递质及Orexin-A水平的影响。[方法]将Wistar大鼠30只随机分为正常对照组、模型对照组和摩腹手法组各10只,采用改良多平台水环境法对后两组大鼠进行睡眠剥夺造模,造模后正常对照组和模型对照组大鼠不作干预,摩腹手法组大鼠给予每日1次的摩腹治疗,持续14 d后,检测比较各组大鼠脑内5-羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NE)、γ-氨基丁酸(GABA)和食欲素A(Orexin-A)的水平。[结果]与正常对照组相比,模型对照组大鼠脑内5-HT、DA、NE、GABA和Orexin-A的水平均发生显著变化(P0.05)。与模型对照组相比,摩腹手法组大鼠脑内5-HT水平显著回升,DA、NE和Orexin-A水平均明显回落(P0.05),而GABA水平无统计学差异(P0.05)。[结论]摩腹手法可通过调节大鼠脑内5-HT、DA、NE和Orexin-A的水平,起到调节动物睡眠和觉醒状态的作用。     

Localization of orexin-A-like immunoreactivity in prolactin cells in the bullfrog (Rana catesbeiana) pituitary

Immunohistochemical techniques were employed to investigate the distribution of orexin-A-like immunoreactivity in the bullfrog (Rana catesbeiana) pituitary. Orexin-A-immunoreactive cells were scattered throughout the pars distalis. We found that these cells corresponded to the cells immunostained with antiserum against bullfrog prolactin (fPRL). Immunoelectron microscopic analysis indicated that an orexin-A-like substance coexisted with fPRL within secretory granules. Western blot analysis of bullfrog pituitary extract revealed that anti-human orexin-A antiserum labeled two separate bands which were not labeled with anti-fPRL antiserum. The present study has, for the first time, provided evidence of the intragranular colocalization of orexin-A-like and PRL immunoreactivities in the bullfrog pituitary.     

CSF hypocretin-1 concentrations correlate with the level of fatigue in multiple sclerosis patients

Considering the multiplicity of symptoms associated with multiple sclerosis (MS), there is possibility that hypocretin system function might be involved in the pathogenesis of the disease. The current study aimed to investigate the hypocretin-1 levels in cerebrospinal fluid (CSF) of MS patients in relation to different neurological deficit measures including: Ambulation Index (AI), Expanded Disability Status Scale (EDSS), Fatigue Severity Scale (FSS), and Epworth Sleepiness Scale (ESS) in relapse-onset MS patients. 53 subjects were included into the study: 38 patients with a diagnosis of MS and 15 healthy controls. Among MS patients, 25 had relapsing-remitting and 13 secondary progressive MS. CSF hypocretin-1 levels did not differ between MS patients and healthy controls (p > 0.05). A positive correlation between hypocretin-1 level and fatigue level was found in MS patients (p < 0.05) and this effect was even stronger in the MS subgroup suffering from fatigue (p = 0.01). Hypocretin system seems to be generally unchanged in MS but a positive correlation between hypocretin-1 level and fatigue may indicate involvement of some compensatory mechanisms stimulating the production of the neuropeptide in MS patients.     

迷走神经电刺激对脑外伤后昏迷大鼠前额叶皮质和下丘脑Orexin-A及其受体OX1R表达变化的影响

目的:探讨迷走神经电刺激(VNS)对脑外伤(TBI)昏迷大鼠的促醒作用及可能相关机制。方法:将SD大鼠随机分为3组:空白对照组、假刺激组和刺激组。应用VNS治疗脑外伤后昏迷大鼠,观察其行为学变化,并用ELISA、Western-blot、免疫组织化学技术检测各组大鼠前额叶皮质(PFC)和下丘脑组织的Orexin-A及OX1R表达。结果:刺激组30只大鼠中20只出现翻正反射,假刺激组30只中8只出现翻正反射;将3组的Orexin-A及OX1R含量进行两两比较,发现刺激组Orexin-A及受体OX1R水平高于假刺激组,差异有显著性意义(P0.05)。结论:VNS可提高脑外伤昏迷大鼠意识状态,其可能机制为上调PFC及下丘脑部位Orexin-A及OX1R水平,因此VNS有望成为脑外伤后昏迷促醒有效方法。