无免疫功能低下的肺隐球菌病患者Th1/Th2类细胞因子的变化

目的 分析无免疫功能低下的肺隐球菌病(PC)患者Th1/Th2类细胞因子的变化及其机制.方法 应用酶联免疫吸附测定(ELISA)法测定20例无免疫功能低下的PC患者(PC组)血清中IL-12、γ-干扰素(IFN-γ)和IL-4的浓度,并与20名健康体检者(对照组)进行对比.分离PC组和对照组外周血单个核细胞(PBMC),重组人IL-12 (rhIL-12)刺激48 h后收集上清液,应用ELISA法测定各组培养上清液中IFN-γ和IL-4的浓度.结果 (1)无免疫功能低下的PC患者血清IFN-γ的浓度为(14.5±2.7) ng/L,显著低于对照组的(81.8±9.8) ng/L,差异有统计学意义(t=6.590,P＜0.01),PC组和对照组血清IL-12浓度分别为(2.5±0.5)和(2.5±0.6) ng/L,血清IL-4浓度分别为(6.9±1.3)和(7.3±1.5) ng/L,差异均无统计学意义(t值分别为0.0035和0.2136,均P＞0.05).(2)PC组与对照组PBMC上清液中IFN-γ浓度分别为(55.7 ±13.6)和(51.1±17.5) ng/L,IL-4分别为(5.1±0.7)和(5.0±0.6)ng/L,差异均无统计学意义(t值分别为0.2979和0.0325,均P＞0.05).(3)经rhIL-12刺激后,PC组和对照组PBMC上清液中IFN-γ的浓度均有明显增高,为(4.3±0.5)和(7.9±1.1)倍,PC组增高幅度明显低于对照组,差异有统计学意义(t=3.01,P＜0.01);而IL-4的浓度两组分别增加了(0.9±0.4)和(1.3±0.4)倍,差异无统计学意义(t=0.7240,P＞0.05).结论 无免疫功能低下的PC患者血清Th1类因子(IFN-γ)下降,Th2类因子(IL-4)无明显变化;Th1细胞对IL-12的反应性和敏感性下降可能是血清Th1类因子(IFN-γ)下降的原因之一.     

慢性丙型肝炎和肝硬化患者外周血Th17和Treg细胞及其相关细胞因子水平变化

目的调查慢性丙型肝炎(CHC)和丙型肝炎肝硬化(LC)患者外周血辅助性T淋巴细胞17(Th17)、调节性T淋巴细胞(Treg)及其相关细胞因子水平的变化。方法 2014年10月~2016年10月本院收治的CHC患者33例,丙型肝炎肝硬化患者27例和健康体检者30例。给予CHC患者α-干扰素联合利巴韦林(PR)方案治疗12 m。采用ELISA法检测白介素-6(IL-6)、IL-10、IL-17和转化生长因子-β(TGF-β),使用流式细胞仪检测外周血Th17和CD4+CD25+Foxp3+Treg细胞数。结果 LC和CHC患者外周血Th17和Treg细胞比率和Th17/Treg比值分别为(4.6±0.7)%、(2.7±0.5)%和(1.6±0.4),和(5.8±0.9)%、(3.1±0.6)%和(1.9±0.5),均显著高于健康人[(2.3±0.4)%、(1.7±0.4)%和(1.4±0.2),P0.05],LC患者外周血Th17和Treg细胞比率和Th17/Treg比值显著高于CHC患者(P0.05);LC和CHC患者血清IL-6、IL-10、IL-17和TGF-β水平分别为(9.9±2.5)ng/L、(18.0±3.2)ng/L、(15.3±2.8)ng/L和(5.5±0.8)ng/L,和(14.1±3.0)ng/L、(21.4±4.0)ng/L、(23.3±3.9)ng/L和(7.2±1.2)ng/L,显著高于健康人[(6.7±1.1)ng/L、(14.2±2.9)ng/L、(5.3±0.6)ng/L和(4.3±1.0)ng/L,P0.05],LC患者血清IL-6、IL-10、IL-17和TGF-β水平显著高于CHC患者(P0.05);CHC患者治疗后外周血Th17和Treg细胞比率及血清TGF-β、IL-10和IL-17水平分别为(3.4±0.6)%、(1.9±0.4)%、(4.7±0.6)ng/L、(15.1±2.5)ng/L和(11.5±2.1)ng/L,均显著低于治疗前,差异有统计学意义(t=7.477、t=7.177、t=4.596、t=4.102、t=6.237,P0.05)。结论 CHC和丙型肝炎肝硬化患者外周血Th17和Treg细胞比率增高,相关细胞因子水平也升高,可能参与了丙型肝炎病毒感染的发病过程,在抗病毒治疗后血清细胞因子水平降低,可能有助于对疗效的观察。     

高水平IFN-γ可能是布鲁杆菌病慢性化的特征之一

目的比较急、慢性布鲁氏菌病患者外周血细胞因子IFN-γ、IL-4、TGF-β1、IL-10和IFN-γ/IL-4水平,寻找布病慢性化细胞因子标志物。方法随机抽取乌兰察布市地方病防治中心急、慢性布鲁杆菌病患者各42名作为研究对象,当地健康人群20名作为对照组。采用单因素方差分析对比急性布病患者、慢性布病患者和健康人群IFN-γ、IL-4、TGF-β1、IL-10和IFN-γ/IL-4水平。急性布病患者和慢性布病患者血清IFN-γ、IL-4、TGF-β1、IL-10和IFN-γ/IL-4水平比较采用t检验。结果急、慢性布病患者和健康对照组血清IFN-γ、IL-4、TGF-β1、IFN-γ/IL-4指标比较有统计学意义(F=6.86、11.90、12.25、4.60,P0.01);三组指标IL-10比较无统计学意义(F=2.72,P0.05)。急性布病患者和慢性布病患者血清TGF-β1和IL-10水平比较无统计学意义(t=-1.90,-1.81,P0.05);急性布病患者和慢性布病患者血清IFN-γ、IL-4和IFN-γ/IL-4比较,有统计学意义(t=-1.99、82、-3.3,P0.05)。结论联合TGF-β1和IL-10水平变化,高水平IFN-γ可能是布鲁氏菌病慢性化的生物化学标志物之一。     

高血压并冠状动脉粥样硬化与辅助性T淋巴细胞亚群

背景辅助性T细胞亚群中Th1细胞分泌干扰素(IFN-γ),Th2细胞分泌白介素4(IL-4),有报道认为动脉粥样硬化病人Th1/Th2平衡紊乱.目的 探讨血管紧张素Ⅱ(Ang Ⅱ),IFN-γ和IL-4与高血压患者冠状动脉粥样硬化的关系,研究他们的Th1/Th2功能平衡有无紊乱.方法 临床及冠状动脉造影确诊的高血压合并冠状动脉粥样硬化患者30例为病例组,冠状动脉造影排除冠状动脉粥样硬化的高血压患者22例为对照组.采用放射免疫法检测血浆Ang Ⅱ水平,ELISA法检测血浆中IFN-γ和IL-4水平.结果 1)病例组Ang Ⅱ[( 64.7±30.1) vs 对照组(36.1±12.0)ng/L,P＜0.05]、IFN-γ[(10.3±5.4) vs (4.2±1.5)ng/L,P＜0.01]水平增高;2)病例组IL-4水平无明显变化[(15.7±7.0) vs 对照组(13.5±7.0)ng/L,P＞0.05];3)病例组IFN-γ与IL-4比值较对照组增大(0.72±0.34 vs 0.35±0.17,P＜0.01);4)Ang Ⅱ水平与IFN-γ水平呈正相关(r=0.51,P＜0.01),而与IL-4水平无相关性(r=0.11,P＞0.05).结论 高血压患者的高Ang Ⅱ水平使辅助性T淋巴细胞亚群Th1细胞活性升高,导致辅助性T淋巴细胞亚群Th1/Th2的功能失衡与冠状动脉粥样硬化发生、发展有关.     

Graves病患者外周血Th1、Th2相关细胞因子的表达及意义

目的:探讨Th1/Th2细胞失衡在Graves病发病中的作用.方法:应用酶联免疫吸附法分别检测29例新诊断但未治疗的Graves病患者(GD组)和21例正常对照者(NC组)血清IFN-γ、IL-4(分别代表Th1、Th2分泌的细胞因子)水平,并与血清甲状腺过氧化物酶抗体(TPOAb)、甲状腺球蛋白抗体(TGAb)进行相关分析.结果:GD组患者IFN-γ水平与NC组差异无统计学意义(P＞0.05);GD组患者IL-4水平较NC组高,差异有统计学意义(P＜0.05);GD患者血清IL-4与TPOAb水平呈显著正相关(r=0.419,P＜0.05),但IL-4与TGAb水平(r=0.357,P＞0.05)及IFN-γ与TPOAb/ TGAb水平均无相关性(分别为r=0.180,r=0.222;P＞0.05).结论:新诊断未治疗的Graves患者以分泌Th2型细胞因子为主,其Th1/Th2细胞免疫失衡偏向Th2细胞占优势的免疫应答.     

辅助性T淋巴细胞17在肝囊型包虫病患者外周血中的变化

目的 探讨肝囊型包虫病患者外周血中辅助性T淋巴细胞(Th)17的变化.方法 将56例受试者分为:健康志愿者(HD)组20例,肝囊型包虫病(CE)组18例,肝囊型包虫病合并胆瘘(BF)组18例.流式细胞仪胞内染色分析法检测各组受试者外周血中Th17细胞占CD4+T淋巴细胞比例,ELlSA检测血清Th17细胞相关细胞因子IL-17、IL-23水平.数据行t检验和Pearson相关分析.结果 CE组患者外周血Th17/CD4+T淋巴细胞比例为(0.23±0.11)%,明显低于BF组的(0.76±0.43)%和HD组的(0.52±0.50)%(t=2.225,t=4.077,均P＜0.05),而BF组和HD组间差异无统计学意义(t=1.931,P＞0.05).血清IL-17和IL-23在CE组分别为(12.1±3.7)ng/L和(84.4±46.0)ng/L,明显低于BF组的(15.5±4.1)ng/L和(138.6±37.9)ng/L(t=2.515,t=3.649,均P＜0.05),也低于HD组的(14.8±4.4)ng/L和(138.1±48.7)ng/L(t=2.401,t=3.706,均P＜0.05),而BF组和HD组间差异无统计学意义(t=0.534,P＞0.05).各组血清IL-17水平与IL-23水平呈正相关(r=0.657,P＜0.05).结论 肝囊型包虫病患者外周血中Th17细胞比例明显减少,血清IL-17、IL-23水平显著降低.     

拉米夫定治疗对慢性乙型肝炎患者干扰素-γ和白细胞介素-4的影响

目的 观察拉米夫定治疗慢性乙型肝炎患者血清IFN-γ和IL-4水平的动态变化.方法 ELISA法分别检测66例慢性乙型肝炎患者在拉米夫定治疗前,治疗后第3、6、9、12个月时的血清IFN-γ和IL-4水平.选取20名健康献血员作为健康对照.治疗前后比较用t检验,计数资料采用非参数秩和检验.结果 HBeAg阳性患者拉米夫定治疗前.完全应答组IFN-γ水平为(21.03±4.44)ng/L,明显高于部分应答组的(13.85±3.92)ng/L及无应答组的(10.63±3.11)ng/L(t=7.56,t=11.87,均P<0.01);以IFN-γ为15.66 ng/L为界,治疗前IFN-γ高水平患者完全应答率明显高于低水平患者(31.0%比8.7%,x2=8.391,P<0.01),无应答率明显低于低水平患者(13.8%比52.2%,x2=4.256,P<0.01).治疗后完全应答组患者IFN-γ/IL-4接近或高于对照组,部分应答组和无应答组低于对照组;HBeAg阴性患者拉米夫定治疗后IFN-γ/IL-4缓慢上升,但未达对照组水平.结论 拉米夫定治疗慢性乙型肝炎可增加IFN-γ释放,抑制IL-4释放,治疗应答程度与治疗后辅助性T淋巴细胞(Th)1/Th2平衡的恢复及治疗前IFN-γ水平有关.     

Th1/Th2免疫应答系统在结核性胸膜炎患者中的表达

目的分析结核性胸膜炎患者辅助T细胞(Th)亚型Th1样细胞因子γ干扰素(IFN-γ)、白细胞介素(IL)-2和Th2样细胞因子IL-4、IL-10在血清与胸腔积液中的分布特点,探讨系统及局部的Th1/Th2免疫应答在人类结核性胸膜炎病理生理过程中的可能作用.方法应用夹心酶联免疫吸附测定(ELISA)法检测51例结核性胸膜炎患者(胸膜炎组)血清及胸液标本中IFN-γ、IL-2、IL-4、IL-10的浓度,并与肺结核患者(肺结核组,36例)和健康人(健康人组,24名)比较.结果(1)3组受试者血清IFN-γ、IL-2、IL-4、IL-10浓度中位数分别为胸膜炎组为118.60 ng/L、0.00 ng/L、1.49 ng/L、0.00 ng/L;肺结核组为265.75 ng/L、18.03 ng/L、16.00 ng/L、0.00 ng/L;健康人组为221.70 ng/L、18.52 ng/L、16.00 ng/L、0.00 ng/L.3组受试者血清IFN-γ、IL-2浓度差异无显著性(K-W检验χ2值分别是1.15、4.68,P> 0.05).肺结核组和健康人组受试者血清IL-4、IL-10浓度差异无显著性 (Mann-Whitney 检验Z值分别为-0.27、-1.93, P>0.05),结核性胸膜炎患者血清IL-4浓度明显低于肺结核组(Mann-Whitney检验Z值-2.84,P<0.01).(2)3组受试者血清IFN-γ与IL-4浓度之比依次为胸膜炎组27.93,肺结核组21.72,健康人组10.82,经检验差异无显著性(K-W检验χ2值为4.18,P>0.05).(3)结核性胸腔积液细胞因子浓度(中位数)IFN-γ为832.70 ng/L,IL-2为43.76 ng/L, IL-4为26.00 ng/L, IL-10为38.69 ng/L, 与血清浓度相比较差异有显著性(Wilcoxon Signed Rank检验,Z值分别为-4.34、-2.82、-3.29、-5.15,P<0.05 ).结论免疫功能健全的肺结核和结核性胸膜炎患者其系统的Th1应答相似;而结核性胸膜炎患者系统Th2应答降低,系统Th1/Th2平衡有所上调.结核性胸膜炎患者胸膜局部的Th1/Th2应答较系统明显增强,这可能是结核性胸膜炎病理生理的特征.Th1/Th2免疫应答在结核性胸膜炎病理生理过程中起重要作用,但这种作用的因果关系尚待进一步研究.     

外周血Th1/Th2/Treg细胞及相应细胞因子水平在耐多药结核疾病中的诊断价值及意义

目的外周血Th1/Th2/Treg细胞及相应细胞因子水平在耐多药结核(MDR-TB)患者中的诊断价值及意义。方法回顾性分析自2016年2月-2020年2月于我院接受治疗的肺结核患者99例,其中无耐药肺结核患者52例(无耐药组),耐多药结核患者47例(耐药组),另外抽取同期体检的健康者76例(健康组),采用酶联免疫吸附试验法检测各组Th1细胞及IFN-γ细胞因子,Th2细胞及IL-4细胞因子,Treg细胞及IL-10细胞因子水平。结果与对照组相比,耐药组和无耐药组患者血清Th1细胞及其细胞因子IFN-γ水平、Th2细胞及其细胞因子IL-4水平以及Treg细胞及其细胞因子IL-10水平均显著降低(P均0.001);耐药组患者血清Th1细胞及其细胞因子IFN-γ水平、Th2细胞及其细胞因子IL-4水平以及Treg细胞及其细胞因子TGF-β水平均较不耐药组患者显著低(P均0.05)。结论患者外周血Th1/Th2/Treg细胞及相应细胞因子水平的变化与耐多药结核发病相关,因此可考虑将其作为耐多药结核疾病诊断的参考指标。     

CHB患者Th17-Treg失衡中IL-21的作用及机制

目的研究慢性乙型肝炎(CHB)患者外周Th17/调节性T细胞(Treg)失衡中IL-21的作用及其初步机制。方法流式细胞仪分别检测59例CHB患者和20例健康人外周血细胞中Th17细胞和Treg细胞频数,同时以ELISA法检测血清细胞因子IL-21、IL-17、IL-10、TGF-β水平。体外分别以IL21、IL-2刺激CHB患者外周血单核细胞(PBMC),以实时PCR检测培养细胞转录因子RORγt mRNA及Foxp3 mRNA表达水平,以ELISA法检测培养上清细胞因子IL-17、IL-10和TGF-β水平。结果与正常人群相比,IL-21、IL1 7、TGF-β水平显著升高(P均0.05),CHB患者外周Th17/Treg比例显著升高(P0.05),IL-21水平与Th1 7/Treg比例呈正相关(P=01.008)。与IL-2相比.IL-21刺激CHB患者PBMC的RORγt mRNA表达水平明显升高(P=0.016)而Foxp3 mRNA表达水平明显降低(P=0.018),相对应的培养上清中,IL-1 7水平显著上升(P=0.049)而IL-10、TGF-β水平明显降低(P=0.017,P=0.004)。结论 C、HB患者中IL-21能通过提高RORγt mRNA表达,同时抑制Foxp3 mRNA表达进而促进Th1 7/Treg失衡。     

Three layer functional model and energy exchange concept of aging process

Relying on a certain degree of abstraction, we can propose that no particular distinction exists between animate or living matter and inanimate matter. While focusing attention on some specifics, the dividing line between the two can be drawn. The most apparent distinction is in the level of structural and functional organization with the dissimilar streams of ‘energy flow’ between the observed entity and the surrounding environment. In essence, living matter is created from inanimate matter which is organized to contain internal intense energy processes and maintain lower intensity energy exchange processes with the environment. Taking internal and external energy processes into account, we contend in this paper that living matter can be referred to as matter of dissipative structure, with this structure assumed to be a common quality of all living creatures and living matter in general. Interruption of internal energy conversion processes and terminating the controlled energy exchange with the environment leads to degeneration of dissipative structure and reduction of the same to inanimate matter, (gas, liquid and/or solid inanimate substances), and ultimately what can be called ‘death.’ This concept of what we call dissipative nature can be extended from living organisms to social groups of animals, to mankind. An analogy based on the organization of matter provides a basis for a functional model of living entities. The models relies on the parallels among the three central structures of any cell (nucleus, cytoplasm and outer membrane) and the human body (central organs, body fluids along with the connective tissues, and external skin integument). This three-part structural organization may be observed almost universally in nature. It can be observed from the atomic structure to the planetary and intergalactic organizations. This similarity is corroborated by the membrane theory applied to living organisms. According to the energy nature of living matter and the proposed functional model, the decreased integrity of a human body's external envelope membrane is a first cause of the structural degradation and aging of the entire organism. The aging process than progresses externally to internally, as in single cell organisms, suggesting that much of the efforts towards the restoration and maintenance of the mechanisms responsible for structural development should be focused accordingly, on the membrane, i.e., the skin. Numerous reports indicate that all parts of the human body, like: bones, blood with blood vessels, muscles, skin, and so on, have some ability for restoration. Therefore, actual revival of not only aging tissue of the human body's membrane, but the entire human body enclosed within, with all internal organs, might be expected. We assess several aging theories within the context of our model and provide suggestions on how to activate the body's own anti-aging mechanisms and increase longevity. This paper presents some analogies and some distinctions that exist between the living dissipative structure matter and inanimate matter, discusses the aging process and proposes certain aging reversal solutions.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Unusual responses of nocturnal pineal melatonin synthesis and secretion to swimming: Attempts to define mechanisms

Abstract: The effect of swimming at night on rat pineal melatonin synthesis was compared with that of light exposure at night. Rats were forced to swim at 0030 hr (lights out at 2000 hr) and sacrificed by decapitation 15 and 30 min later, immediately after swimming. Other groups of animals were exposed to white light (650μW/cm²) for 15 and 30 min at same time. Swimming caused a rapid and highly significant drop in the melatonin content in the pineal gland; however, the activity of N-acetyltransferase (NAT), the supposed rate limiting enzyme in the melatonin production, was not changed. Despite the drop in pineal melatonin levels, serum concentrations of the indole remained elevated in the rats that swam. In contrast, melatonin levels in the pineal and serum of light exposed rats fell precipitously, accompanied by a significant suppression of NAT activity. Since we anticipated that the strenuous exercise associated with swimming may induce release of artrial natriuretic peptide (ANP) from the heart, which in turn could cause the release of pineal melatonin, in a second study we injected physiological saline intravenously to stretch the cardiac muscle and release ANP. Three milliliters of normal saline was injected during the day into the jugular vein of anesthetized rats that were pretreated with isoproterenol to stimulate pineal melatonin production. Animals were killed 15 min after the saline injection, and pineal NAT activity and pineal melatonin levels were measured. The saline injections caused no alteration in the elevated levels of either NAT or melatonin. These data suggest that the disparity in pineal NAT activity (which was high) and pineal melatonin (which was low), in animals swum at night, may not be caused by ANP which is released during strenuous exercise such as swimming.     

Melatonin and photoperiodic time measurement: Seasonal breeding in the sheep

Abstract: Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status—the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.