泌乳素及其受体的研究进展

泌乳素(PRL)又名催乳素,是由腺垂体及一些垂体外器官如乳腺、胸腺、脾脏等合成的一种多肽类激素,以内分泌、自分泌、旁分泌的形式发挥作用,其广泛参与机体生长发育、物质代谢、性腺功能调节、应激反应、免疫调节等。泌乳素通过与其受体(PRL receptor,PRLR)在靶细胞的细胞膜表面结合,激活下游的信号转导通路,发挥其生物学作用。由于泌乳素在人体内复杂的生物学效应,泌乳素及其受体又与乳腺癌、泌乳素瘤等多种疾病的发生发展及其预后密切相关。本文就泌乳素发挥效应时对其受体的激活、受体激活后的信号转导机制以及泌乳素同相关疾病的联系进行了综述,相信泌乳素及其受体的研究将为这些疾病的治疗提供重要的方向。     

疏肝健脾法对产后缺乳大鼠催乳素及其受体表达的影响研究

目的:观察疏肝健脾法对产后缺乳肝郁脾虚证大鼠乳腺组织形态学与催乳素(prolactin,PRL)及其受体(prolactin receptor,PRLR)表达的影响。方法:采用甲磺酸溴隐亭溶液+慢性束缚法复制大鼠产后缺乳肝郁脾虚病证结合动物模型,随机分为模型组、空白对照组、西药对照组、中药低、中、高剂量组,每组8只,共6组。药物干预后,采用HE染色及免疫组化法比较不同干预方法对产后缺乳大鼠乳腺组织形态与催乳素及其受体表达的影响,匀浆乳腺组织,采用ELISA法检测乳腺组织催乳素(PRL)、血清催乳素(PRL)、胃泌素(GAS)、血清下丘脑一垂体一肾上腺轴(The hypothalamic-pituitary-adrenal axis,HPA轴)水平变化:促肾上腺皮质激素释放激素(CRH)、促肾上腺皮质激素(ACTH)、皮质醇(COR)。结果:与模型组相比,西药对照组和中药低、中、高剂量组均可促进乳腺组织向泌乳活跃期发育,表现为腺泡腔充盈、腺泡上皮细胞数量增多、结缔组织和脂肪组织退化,同时增加乳腺组织中PRL(空白对照组(145.72±16.332)、中药中剂量组(127.39±10.854)西药对照组(80.79±18.112)、中药低剂量组(89.42±14.183)、中药高剂量组(88.02±12.064)模型组(54.32±7.128))(P0.05)及PRLR(空白对照组(139.76±14.199)、中药中剂量组(129.09±18.755)西药对照组(87.42±8.390)、中药低剂量组(81.54±11.090)、中药高剂量组(85.167±9.494)模型组(60.93±11.767))的表达(P0.05)。与模型组相比,疏肝健脾中药可降低过高的血清CRH、ACTH、COR水平,升高乳腺组织PRL、血清GAS及PRL水平(P0.05)。其中,中药中剂量组疗效优于其他用药各组(P0.05)。结论:疏肝健脾法促进产后缺乳大鼠的泌乳作用机制与增加PRL及其受体表达、调节胃肠功能、抑制HPA轴的亢进状态,从而促进乳腺组织形态向泌乳活跃期发育密切相关。     

心理应激致孕鼠乳腺发育不良及雌/孕激素和受体水平异常

目的研究心理应激对孕鼠乳腺发育的影响,评价相关激素及其受体的表达水平。方法妊娠期Wistar大鼠随机分为对照组和实验组,实验组大鼠15 d内随机施加噪音、束缚、昼夜颠倒、冰水游泳和夹尾致痛5种不可预见性应激刺激。观察大鼠乳房外观变化,测量乳头直径和高度并计算乳房系数,光密度法测定乳腺组织DNA和RNA水平,放射免疫法测定血浆和乳腺组织雌激素(E2)、孕酮(P)、生长激素(GH)及催乳素(PRL)水平,放射配基结合分析法测定雌二醇、孕酮受体的最大结合容量(Bmax)和解离常数(Kd),乳房组织H-E染色,光镜下观察组织形态变化。结果与对照组比较,实验组大鼠乳头直径和高度减小、乳房重量减轻、乳房系数减小(P0.05,P0.01),乳房组织DNA、RNA及RNA/DNA均明显减小(P0.05,P0.01),血浆E2、P和GH水平明显下降(P0.05,P0.01),组织匀浆E2和GH水平明显降低(P0.05,P0.01),E2受体和P受体Bmax明显降低、Kd均明显升高(P0.05,P0.01);乳腺小叶、小叶腺泡数目减少、腺泡直径明显降低(P0.05)。结论心理应激能导致孕鼠乳腺发育不良,引起雌、孕激素水平及其受体表达异常。     

舒芬太尼联合地佐辛用于剖宫产术后镇痛的疗效及对患者血清泌乳素水平的影响

目的:探讨舒芬太尼联合地佐辛对剖宫产患者术后镇痛的效果及对患者血清泌乳素水平的影响。方法:选取我院收治的剖腹产患者90例,随机分为对照组及实验组,对照组给予舒芬太尼联合托烷司琼术后镇痛,实验组给予舒芬太尼联合地佐辛术后镇痛。观察并比较两组患者的镇痛效果及血清泌乳素PRL水平的影响,以及不良反应的发生情况等。结果:与术前比较,两组患者治疗后血清泌乳素PRL水平均升高,差异具有统计学意义(P0.05);与对照组比较,实验组治疗后血清PRL水平较高,差异具有统计学意义(P0.05);与对照组比较,实验组镇痛VAS评分、初次泌乳时间及泌乳充足时间、不良反应发生率均较低,差异具有统计学意义(P0.05)。结论:剖宫产术后应用舒芬太尼联合地佐辛的临床镇痛效果理想,在维持镇痛效果的同时,降低药物用量,减轻对脑垂体的刺激,提高泌乳素分泌,有效降低不良反应,值得临床推广应用。     

SSTR-2、E-cad与不明原因的反复自然流产相关性研究

目的:研究生长抑素2型受体(SSTR2)、E-钙粘附素(E-cad)与不明原因的反复自然流产相关性。方法:选取2011年1月到2014年12月我院收治的不明原因反复自然流产患者60例(研究组),另选同期早期正常孕妇且准备流产者60例(对照组),应用免疫组化技术检测两组蜕膜组织中的SSTR2和E-cad表达,并进行比较。结果:研究组蜕膜组织中SSTR-2阳性细胞平均光密度显著高于对照组,而E-cad阳性细胞平均光密度显著低于对照组,两组比较差异具有统计学意义(P0.05);研究组蜕膜组织中SSTR-2与E-cad水平呈负相关关系,对照组蜕膜组织中SSTR-2与E-cad水平呈正相关关系(P0.05)。结论:蜕膜组织中SSTR-2高表达与E-cad低表达可能与不明原因的反复自然流产有关。     

中药当归枸杞子汤调节高泌乳素血症临床观察

目的通过连续动态监测泌乳素血液浓度,评价当归枸杞子汤对高泌乳素血症的调节作用。方法选择我院2012年5月~2013年5月门诊收治的高泌乳素血症患者130例,随机分为对照组30例,采用溴隐亭治疗;观察组100例,给予中药当归枸杞子汤治疗。连续动态监测血清泌乳素浓度,评价两组疗效。结果两组患者疗效比较无显著统计学意义(P0.05)。两组治疗后血清泌乳素均显著下降,与治疗前比较差异有统计学意义(P0.05);而两组治疗后比较差异无统计学意义(P0.05)。不良反应发生率比较,对照组显著高于观察组(P0.05)。结论中药当归枸杞子汤治疗高泌乳素血症安全、有效,且价格低廉,用法简单,有显著的临床推广应用价值。     

谷物发酵物促进产后缺乳小鼠泌乳功效的机制研究

摘要 目的：研究谷物发酵物对产后缺乳小鼠促进泌乳的功效及作用机制。方法：选择分娩时间相差不超过24 h的KM小鼠,随机分为缺乳模型组、正常组、雌二醇组（17α-雌二醇,1 g/kg）和谷物发酵物组（24 mL/kg）,每组6只。产后第3 d起,除正常组外,其余各组均采用溴隐亭（1.6 mg/kg）建立产后缺乳模型,共10 d。观察仔鼠、母鼠体重变化,每日泌乳量变化,ELISA法检测血浆催乳素（PRL）水平、HE染色观察乳腺组织学形态变化,免疫组化法检测催乳素受体（PRLR）、β-酪蛋白（β-casein）在乳腺组织中的表达。结果：正常组、雌二醇组及谷物发酵物组仔鼠窝重增加量,泌乳量,血浆PRL水平,乳腺中PRLR和β-casein的表达均高于缺乳模型组(P＜0.05) ,母鼠体重减少量低于缺乳模型组(P＜0.05),谷物发酵物组与缺乳模型组相比乳腺小叶面积明显变大,小叶间结缔组织和脂肪组织明显减少,腺泡大量增生,小叶内腺泡腔恢复到正常水平以上。结论：谷物发酵物对溴隐亭诱导的产后缺乳母鼠有促进泌乳作用,泌乳作用机制可能与增加PRL及其受体、β-casein表达有关。     

替莫唑胺治疗垂体瘤患者的临床疗效及机制研究

目的:探讨替莫唑胺治疗垂体瘤患者的临床疗效及其机制。方法:选取我院收治的垂体瘤患者70例,随机分为实验组和对照组,每组35例。对照组患者进行显微镜下经鼻蝶窦手术治疗,并给予溴隐亭口服治疗;实验组在对照组治疗基础上联合替莫唑胺胶囊治疗。观察并比较治疗前后两组患者血清泌乳素、IGF-1、GH水平的变化及临床疗效。结果:与治疗前相比,两组患者治疗后的血清PRL、IGF-1、GH、TSH水平均降低,FT4、ACTH水平升高,差异具有统计学意义(P0.05);而与对照组比较,实验组的血清PRL、IGF-1、GH、TSH水平更低,FT4、ACTH水平更高(P0.05)。实验组的治疗总有效率显著高于对照组,差异具有统计学意义(P0.05)。结论:替莫唑胺治疗垂体瘤的临床疗效好,推测其机制与降低血清PRL、IGF-1水平有关。     

葛根素对正常产后小鼠泌乳作用的影响

摘要 目的：探究葛根素对产后正常小鼠泌乳作用的影响及其机制,并初步探究葛根素对产后正常小鼠的安全性。方法：将雌、雄KM小鼠以3:1比例合笼配种,得到孕鼠饲养至分娩。分娩后的小鼠随机分为正常对照组、葛根素低剂量（18 mg?kg^-1）、高剂量组（72 mg?kg^-1）,每组8只。从产后第3 d起,每天灌胃一次,共10 d。观察小鼠每日泌乳量变化,ELISA法检测血清中催乳素（PRL）、孕酮（P4）、雌二醇（E₂）含量,HE染色观察乳腺、肝、肾、子宫、卵巢组织病理学形态,Western Blot法检测乳腺组织中催乳素受体（PRLR）、酪氨酸激酶 2（JAK2）和信号传导与激活因子5a（STAT5a）的表达。结果：与正常对照组相比,从给药的第6天起,葛根素低剂量组的泌乳量显著升高（P＜0.05）;葛根素低、高剂量组均可见乳腺小叶内腺泡明显变大,分泌物明显增多,且低剂量组更为明显;葛根素低、高剂量组血清PRL水平明显升高（P＜0.01或P＜0.05）;葛根素低剂量组PRLR的蛋白表达明显增加（P＜0.01）,而葛根素高剂量组PRLR、JAK2的蛋白表达明显降低（P＜0.01）。葛根素低剂量组PRLR、JAK2、STAT5a的蛋白表达明显高于葛根素高剂量组（P＜0.01或P＜0.05）。结论：葛根素低剂量对产后正常小鼠有一定促进泌乳作用,高剂量时对泌乳作用不明显。葛根素低、高剂量均未对产后正常小鼠的肝、肾、卵巢和子宫产生明显的病理学改变。     

磁共振成像表观扩散系数与乳腺浸润性导管癌组织学分级及预后指标的相关性研究

目的:探讨磁共振成像(MRI)表观扩散系数(ADC)与乳腺浸润性导管癌组织学分级及其预后指标的相关性。方法:收集2016年5月至2017年5月于我院就诊的并经手术病理确诊为乳腺浸润性导管癌的患者112例作为研究对象,选取患者乳腺癌组织样本作为病例组,同时选取患者对侧正常乳腺组织样本作为对照组,所有患者均行常规MRI和磁共振扩散加权成像(DW-MRI)检查,分别测量两组样本的ADC值,比较不同乳腺浸润性导管癌组织学分级与正常乳腺组织的ADC值,分析乳腺浸润性导管癌组织的ADC值与肿瘤直径大小、淋巴结转移状态、有无远处转移及雌激素受体(ER)、孕激素受体(PR)和Ki-67表达的关系,并分析ADC值与组织学分级及预后指标的相关性。结果:乳腺浸润性导管癌病理分级I级的ADC值低于对照组,病理分级II级的ADC值低于病理分级I级及对照组,病理分级III级的ADC值低于病理分级II级、I级及对照组,差异均具有统计学意义(P0.05)。乳腺浸润性导管癌患者中,肿块直径2 cm、无淋巴结转移、ER阴性、PR阴性、Ki-67阴性患者的平均ADC值均高于肿块直径≥2 cm、有淋巴结转移、ER阳性、PR阳性、Ki-67阳性患者,差异均具有统计学意义(P0.05);而有无远处转移患者之间比较差异无统计学意义(P0.05)。经Spearman秩相关分析结果显示,乳腺浸润性导管癌患者的ADC值与病理组织学分级呈现负相关关系(rs=-0.716,P=0.000);与肿块直径大小、有无淋巴结转移及ER、PR、Ki-67的表达均呈负相关(rs=-0.316、-0.545、-0.667、-0.598、-0.443,P均0.05),与有无远处转移无相关性(rs=0.091,P=0.887)。结论:乳腺浸润性导管癌的ADC值与癌组织学分级和预后相关指标存在一定相关性,可作为一种临床诊断和判断预后的重要指标,具有重要临床价值。     

Effects of the SLICK1 mutation in PRLR on regulation of core body temperature and global gene expression in liver in cattle

The SLICK1 mutation in bovine PRLR (c.1382del; rs517047387) is a deletion mutation resulting in a protein with a truncated intracellular domain. Cattle carrying at least one allele have a phenotype characterized by a short hair coat (slick phenotype) and increased resistance to heat stress. Given the pleiotropic nature of prolactin, the mutation may affect other physiological characteristics. The liver is one organ that could potentially be affected because of the expression of PRLR. The mutation is a dominant allele, and heterozygous animals have a similar hair coat to that of animals homozygous for the mutation. Present objectives were to determine whether inheritance of the SLICK1 mutation affects liver gene expression and if animals homozygous for the SLICK1 allele differ from heterozygotes in liver gene expression and regulation of body temperature during heat stress. In one experiment, rectal and ruminal temperatures were less for Holstein heifers that were heterozygous for the SLICK1 allele compared with wildtype heifers. There were 71 differentially expressed genes in liver, with 13 upregulated and 58 downregulated in SLICK1 heterozygotes. Among the ontologies characteristic of differentially expressed genes were those related to immune function and fatty acid and amino acid metabolism. In a prospective cohort study conducted with adult Senepol cattle, body temperature and hepatic gene expression were compared between animals heterozygous or homozygous for the SLICK1 mutation. There were no differences in ruminal temperatures between genotypes, rectal temperature was higher in animals homozygous for the SLICK1 mutation, and there was only one gene in liver that was differentially expressed. It was concluded that inheritance of the SLICK1 allele can exert functional changes beyond those related to hair growth although changes in liver gene expression were not extensive. Results are also consistent with the SLICK1 allele being dominant because there were few differences in phenotype between animals inheriting one or two copies of the allele.     

Arginine vasotocin and renal function in the conscious dog

The effects of arginine vasotocin (AVT), 4–6 ng/kg, on renal function and plasma prolactin (PRL) were determined in conscious American foxhounds undergoing a water diuresis. Intravenous AVT produced an antidiuresis, kaliuresis and inconsistent natriuresis, independently of changes in GFR, blood pressure, heart rate and plasma PRL.     

草鱼催乳素抗血清的制备与鉴定

应用从草鱼垂体中分离纯化的催乳素免疫兔子制备了特异抗血清。用常规酶联免疫吸附测定法检测表明该抗血清与马哈鱼生长激素及促性腺激素完全没有交叉反应，而与草鱼生长湟级向弱交叉反应。免疫细胞化学染色表明该抗血清主要与草鱼垂体前叶催乳素细胞发生结合反应，与垂体间叶细胞有微弱染色反应，但与神经垂体琢垂体后叶细胞则完全没有免疫结合反应。     

Stress-induced hormonal and mood responses in scuba divers: a field study

The majority of injuries in scuba-divers are attributable to inappropriate behavior under stressful diving conditions, predominantly involving panic reactions emerging from elevated levels of anxiety. Divers with an elevated level of anxiety and poor coping are at higher risk of developing panic reactions than those possessing more adequate stress-coping-mechanisms. In the comparison of two extreme groups of seven divers each with opposite stress coping strategies, prolactin was found to be a hormonal marker with a significant increase in the sub-group of the stress-controllers. This hormonal response was observed in a recreational and a stressful dive, and in the latter with a more distinct elevation. Along with the self-reported emotional conditions under immersion, these data suggest that an increased prolactin level reflects a state of elevated physical and mental activation and vigilance. Facing a stressful situation subjects with more emotional concern and the tendency to surrender react by "blunted responses" and show significantly lower elevations of the prolactin levels in contrast to subjects with the very opposite psychological features. The other observed somatic parameters (epinephrine, norepinephrine) showed significant increases during and after dives (with the exception of saliva cortisol), however without any significant group difference.     

Purification of Monomeric Prolactin Charge Isoform from Buffalo Pituitaries

Abstract

A standard protocol for isolation of buffalo prolactin (buPRL) was modified at the alcohol precipitation step. This modification could separate lower molecular weight prolactin from the higher molecular weight prolactin (PRL). Reloading the prolactin onto a Sephacryl S‐200 gel purified the buPRL monomer. The purity of buPRL monomer was confirmed by 15% SDS PAGE. The buPRL monomer was >90% pure. It was characterized by specific anti‐buPRL serum in ELISA and Western blot. A native PAGE of the PRL showed three charge isoforms. A protocol was standardized to separate prolactin monomeric least acidic isoforms using an anion exchanger.     

Prolactin-releasing activity of porcine intestinal peptide (PHI-27)

Porcine intestinal peptide (PHI), a twenty-seven amino acid peptide isolated from porcine gut extracts, is a close structural homolog of the secretin family hormones. The structural and biological similarities of PHI to vasoactive intestinal peptide (VIP) together with its presence in the rat hypothalamus suggested a possible role for the peptide in the control of prolactin (PRL) secretion. PHI induced significant, dose-related stimulations of PRL release from cultured, dispersed rat pituitary cells in vitro. The minimum effective dose is 10⁷ molar, compared to 10⁹ molar for VIP. No interactive effect with thyrotropin-releasing hormone was observed; however, PHI partially overcame the dopamine inhibition of PRL release.     

转基因小鼠乳腺上皮细胞的体外培养及其对催乳素反应的研究

实现转基因生物乳腺反应器对外源蛋白的高效表达是目前生物制药亟待解决的难题。催乳素对泌乳期乳蛋白的合成与分泌具有重要的调控功能。通过转基因小鼠乳腺上皮细胞模型的建立,研究催乳素如何调控乳蛋白的表达,为提高乳腺反应器高效表达外源蛋白提供技术及理论支撑。应用机械破碎及胶原酶消化法,经差速贴壁纯化,成功培养含人转铁蛋白基因的小鼠乳腺上皮细胞,细胞上清液中检测到人转铁蛋白表达。细胞经牛催乳素诱导后人转铁蛋白的表达水平明显升高。利用转基因小鼠乳腺上皮细胞模型,可以进行催乳素和环境因素等对乳腺上皮细胞合成及分泌蛋白能力影响的研究。