Corticotropin-releasing factor stimulates secretion of melanocyte-stimulating hormone from the rat pituitary

Administration of synthetic ovine corticotropin-releasing factor led to rapid, parallel increases in adrenocorticotropin and alpha-melanocyte-stimulating hormone concentrations in rat plasma. Prior treatment with dexamethasone almost completely blocked the adrenocorticotropin response but not the increase in melanocyte-stimulating hormone. These data demonstrate that corticotropin-releasing factor is a potent stimulator not only of adrenocorticotropin secretion from the corticotrophs of the anterior pituitary gland but also of peptide secretion from the intermediate lobe. Such data suggest that melanocyte-stimulating hormone and beta-endorphin play a role in the physiological response to stress.     

Dexamethasone fails to suppress beta-endorphin plasma concentrations in humans and rhesus monkeys

In humans and rhesus monkeys, dexamethasone decreased concentrations of plasma cortisol but did not alter circulating beta-endorphin immunoreactivity. Contrary to current theory suggesting that pituitary beta-endorphin and adrenocorticotropic hormone are controlled by identical regulatory mechanisms for synthesis and release, our evidence suggests that in higher primates the established glucocorticoid feedback mechanism for the adrenocorticotropic hormone-cortisol system does not regulate beta-endorphin secretion in the same way.     

Evidence for homologous actions of pro-opiocortin products

alpha-Melanocyte-stimulating hormone (alpha-MSH), a modified fragment of adrenocorticotropic hormone, derives from the same biosynthetic route as beta-endorphin and is stored by the same arcuate neurons. Microinjection of alpha-melanocyte-stimulating hormone and several related peptides into the periaqueductal gray matter significantly reduced responsiveness to pain and had a behavioral profile similar to that produced by beta-endorphin.     

中国瘰螈皮肤转录组测序及活性多肽分析

首次对中国瘰螈皮肤组织进行了转录组测序并分析其活性多肽。提取中国瘰螈皮肤组织总RNA用于构建cDNA文库，采用Illumina HiSeq 4000进行测序。测序数据处理后获得unigene，并对其进行基因注释和生物信息学分析。共组装获得了74 371 个非冗余unigene（平均长度为762 bp），其中33 627个unigene在七大功能数据库中得到功能注释，5 137个unigene的注释结果来自两栖类物种，约占注释总数的15%。另外，从转录组数据中识别了一些皮肤活性多肽如胆囊收缩素、防御素、伤口愈合肽、蛋白激酶抑制剂、胸腺素，它们大多数具有皮肤免疫或防御功能，且在其他两栖动物种类中均发现有较高序列相似性的同源多肽。结果为阐明蝾螈皮肤免疫或防御机制提供了前期基础资料。     

酶法制备大豆蛋白成骨活性肽

【目的】探究从大豆分离蛋白双酶分步酶解产物中分离促成骨细胞增殖肽的工艺,为大豆产品的开发提供参考。【方法】以促成骨细胞增殖活性作为测定指标,选用木瓜蛋白酶酶解大豆分离蛋白（SPI）,采用MTT法检测不同浓度的木瓜蛋白酶酶解产物对成骨细胞的促增殖活性。采用pH-Stat法检测水解时间为1、2、3、4、5和6 h的木瓜蛋白酶酶解产物的水解度,并检测相应水解时间的酶解产物对成骨细胞的促增殖活性。然后分别采用截留分子量为30 kD和10 kD两种超滤膜对活性较高的酶解产物进行超滤,并测定各超滤组分对成骨细胞的促增殖活性。再将具有较高促成骨细胞增殖活性的超滤组分分别在0.5、1、1.5、2和2.5 h进行碱性蛋白酶酶解,并检测相应水解时间的酶解产物对成骨细胞的促增殖活性。然后选用交联葡聚糖（Sephadex G-15）对具有较高促成骨细胞增殖活性的酶解产物进行分离,并检测各分离组分对成骨细胞的促增殖活性。最后,对各分离组分进行氨基酸分析,并采用质谱（ESI-TOF MS/MS）对最高促成骨细胞增殖活性的分离组分进行结构鉴定,并筛选出具有较强促成骨细胞增殖的大豆活性肽。【结果】在浓度为200 μg·mL ^-1时,木瓜蛋白酶酶解物对成骨细胞的促增殖活性随着酶解时间的增加逐渐增强,当酶解时间为5 h时达到最高促增殖活性（（118.24±2.73）%）。木瓜蛋白酶酶解产物经超滤分离、碱性蛋白酶酶解和凝胶过滤色谱分离后,对各分离组分进行氨基酸分析并筛选得到对成骨细胞具有最强增殖活性的组分F3,该组分的成骨细胞增殖率为（125.80±2.94）%,且F3中丙氨酸、缬氨酸、异亮氨酸、亮氨酸、苯丙氨酸等疏水性氨基酸以及芳香族氨基酸和必需氨基酸的含量明显高于其他组分。进一步通过质谱鉴定出F3主要由10个肽段组成。其中,DAMDGWFRL、GQTPLFPR、ADFYNPK、KDWYDIK的疏水性氨基酸、芳香族氨基酸以及必需氨基酸占比较高。对上述4个肽段进行活性测定,发现GQTPLFPR的促成骨细胞增殖活性最强,在浓度为100 μmol·L^-1时,其成骨细胞增殖率为（129.11±3.12）%。【结论】采用双酶分步酶解结合超滤和凝胶过滤色谱等蛋白分离纯化技术,从大豆分离蛋白中分离纯化出了具有较强促成骨细胞增殖活性的多肽,为促成骨细胞增殖活性肽的制备和应用提供了技术参考。     

米糠蛋白抗氧化肽的制备及初步分离

试验以米糠蛋白为原料,制备米糠蛋白抗氧化肽。以DPPH自由基清除率为指标,采用正交试验,优化碱性蛋白酶酶解米糠蛋白制备抗氧化肽的工艺条件;采用超滤、SephadesG-25柱层析分离纯化米糠蛋白抗氧化肽。试验结果表明,制备米糠蛋白抗氧化肽的最优工艺条件为：酶解温度45℃,pH9．0,时间60rain,米糠蛋白底物浓度3％,碱性蛋白酶加酶量3000U·g-1。在米糠蛋白酶解产物中,Mrs〈5KDa组分的抗氧化活性最强,其对DPPH自由基清除率为68．7％。通过SephadexG-25凝胶层析柱进一步分离得到4个混合组分,混合组分Ⅱ活性最强,其DPPH自由基清除率为75．83％。     

磷酸盐缓冲液的浓度和pH对革胡子鲶肠道抗菌蛋白/肽分级盐析的影响

[目的]探讨磷酸盐缓冲液的pH和浓度在分级盐析时,对革胡子鲶肠道各组分蛋白/肽的影响。[方法]以不同浓度和pH的磷酸盐缓冲液作为组织匀浆缓冲液,在硫酸铵饱和度为20%、40%、60%、80%和100%的条件下,对革胡子鲶肠道抗菌蛋白/肽进行分级盐析,并对盐析产物进行称重、电泳检测和抑菌活性分析。[结果]磷酸盐缓冲液的pH为6.0或7.4时,其浓度对盐析产物产量、蛋白沉淀和抑菌效果无显著影响。而pH为8.0、浓度为0.05 mol/L时,硫酸铵饱和度为60%和100%的粗蛋白/肽的产量显著高于浓度为0.02 mol/L的磷酸盐缓冲液粗蛋白含量;大分子蛋白能更好地在硫酸铵饱和度为20%和40%时沉淀下来,而在饱和度为60%、80%和100%能获得分子量相对较小的抗菌蛋白/肽。磷酸盐缓冲液的pH和浓度对抑菌效果无显著影响。[结论]pH8.0,浓度为0.05 mol/L的磷酸盐缓冲液更适于革胡子鲶肠道抗菌蛋白/肽的提取。     

Comparison of the distribution of dynorphin systems and enkephalin systems in brain

A study of the anatomical distribution of the endogenous opioid dynorphin in rat brain showed that the peptide is localized in a widespread system with multiple cell groups and projections. This network is revealed by the use of multiple antiserums against dynorphin and can be distinguished from the system containing methionine-enkephalin and leucine-enkephalin, which is mapped by the use of antiserums against the enkephalins and biosynthetically related peptides in the adrenal. It thus appears that the brain contains at least three separate opioid neuronal networks: an enkephalin family with components similar to those found in the adrenal, a beta-endorphin family, and a dynorphin family.     

Immunoreactive dynorphin-(1-8) and corticotropin- releasing factor in subpopulation of hypothalamic neurons

Immunoreactive corticotropin-releasing factor (CRF) and dynorphin-(I-8) were visualized in rat hypothalamus by immunohistofluorescence with specific antibodies. In brains from colchicine-treated, adrenalectomized rats, neuronal perikarya with immunoreactive CRF were observed in the paraventricular nucleus of the hypothalamus. The CRF occurred together with the dynorphin-(1-8). However, the CRF immunoreactivity occurred only in a subpopulation of the dynorphin-(1-8) immunoreactive cells. These findings suggest that there may be a functional interrelationship of CRF with dynorphin-related opioid peptides and provide further evidence that neurons may contain more than one bioactive substance.     

胸腺肽对幼龄雄性小鼠生殖器官指数及性激素水平的影响

为了探讨胸腺肽对幼龄雄性小鼠生殖器官指数及性激素水平的影响,将100只幼龄雄性小鼠随机分为5组,第1组为正常饲养对照组,不做任何处理;第2组为生理盐水对照组,注射生理盐水;其余3组为试验组,每日分别背部皮下注射低剂量胸腺肽(0.30mg.mL-1)、中剂量胸腺肽(0.60mg.mL-1)和高剂量胸腺肽(1.20mg.mL-1),连续30d后测定睾丸指数、附睾指数、精囊腺指数和血清中促黄体生成素(LH)、睾酮(T)水平。试验结果表明:与生理盐水对照组相比,高剂量胸腺肽小鼠睾丸指数、附睾指数、精囊腺指数及LH水平均显著性升高(p<0.05);低胸腺肽组精囊腺指数显著性升高(p<0.05)。与正常饲养对照组比较,高剂量胸腺肽组小鼠T水平显著性升高(p<0.05)。可见,胸腺肽能提高雄性小鼠生殖器官指数和性激素水平,从而调节雄性小鼠生殖内分泌系统。     

胸腺素β4研究进展

胸腺素βA真核生物细胞中的一种主要的肌动蛋白螯合分子,广泛分布于哺乳动物和其它脊椎动物的多种组织和有核细胞中。尽管其分子水平的作用机制尚不明确,但胸腺素β4却与人类的许多生理及病理过程密切相关。随着研究的进一步深入,胸腺素β4在临床上的潜在应用价值将被开发,这对于一些疾病的诊断、治疗及预防均具有重要意义。近年来对胸腺素β4的研究包括其在创伤愈合、肿瘤转移、血管再生等方面的生物学作用,同时,随着对胸腺素β4需求的增加,对利用基因工程的方法进行胸腺素β4生产的研究也在不断进行中。丈中拟对近年来胸腺素β4在生物学功能和生产方法上所取得的进展进行综述。     

beta-Endorphin: possible involvement in the antihypertensive effect of central alpha-receptor activation

Clonidine and L-alpha-methylnoradrenaline (but not D-alpha-methylnoradrenaline) increase the release of a substance with beta-endorphin immunoreactivity from slices of brainstem of spontaneously hypertensive rats, but not that of normotensive rats. It was reported earlier that opiate antagonists inhibit the hypotensive action of clonidine and alpha-methyldopa in spontaneously hypertensive but not in normotensive rats and that beta-endorphin has hypotensive effects of its own. Together, these findings indicate that release of beta-endorphin by central alpha-receptor agonists may contribute to the antihypertensive action of these drugs.     

Synthetic CD4 peptide derivatives that inhibit HIV infection and cytopathicity

Synthetic peptide segments of the CD4 molecule were tested for their ability to inhibit infection of CD4+ cells by the human immunodeficiency virus (HIV) and to inhibit HIV-induced cell fusion. A peptide mixture composed of CD4(76-94), and synthesis side products, blocked HIV-induced cell fusion at a nominal concentration of 125 micromolar. Upon high-performance liquid chromatography, the antisyncytial activity of the peptide mixture was found not in the fraction containing the peptide CD4(76-94) itself, but in a side fraction containing derivatized peptide products generated in the automated synthesis. Derivatized deletion and substitution peptides in the region CD4(76-94) were used to demonstrate sequence specificity, a requirement for benzyl derivatization, and a core seven-residue fragment required for antisyncytial activity. A partially purified S-benzyl-CD4(83-94) peptide mixture inhibited HIV-induced cell fusion at a nominal concentration of less than or equal to 32 micromolar. Derivatized CD4 peptides blocked cell fusion induced by several HIV isolates and by the simian immunodeficiency virus, SIV, and blocked infection in vitro by four HIV-1 isolates with widely variant envelope gene sequences. Purified CD4(83-94) dibenzylated at cysteine 86 and glutamate 87 possessed antisyncytial activity at 125 micromolar. Derivatization may specifically alter the conformation of CD4 holoreceptor peptide fragments, increasing their antiviral efficacy.     

发酵牛肉火腿中ACE抑制肽的稳定性研究

研究了发酵牛肉火腿中提取的ACE抑制肽的热处理、室温储存和模拟胃肠道消化稳定性。结果表明,发酵牛肉火腿中提取的ACE抑制肽,在50、72、85和100℃下处理5 min和100℃下5、10、20和40 min处理后,处理组和对照组ACE抑制活性无显著差异。在室温下储存2 d,处理组和对照组ACE抑制活性无显著差异(P0.05),但储存至3 d,它的ACE抑制活性显著低于对照组(P0.05),其后储藏时间对提取物的ACE抑制活性无显著影响。ACE抑制肽经体外模拟胃肠道消化后,其活性与对照组无显著差异。     

Regulation of CD8+ T cell development by thymus-specific proteasomes

Proteasomes are responsible for generating peptides presented by the class I major histocompatibility complex (MHC) molecules of the immune system. Here, we report the identification of a previously unrecognized catalytic subunit called beta5t. beta5t is expressed exclusively in cortical thymic epithelial cells, which are responsible for the positive selection of developing thymocytes. Although the chymotrypsin-like activity of proteasomes is considered to be important for the production of peptides with high affinities for MHC class I clefts, incorporation of beta5t into proteasomes in place of beta5 or beta5i selectively reduces this activity. We also found that beta5t-deficient mice displayed defective development of CD8(+) T cells in the thymus. Our results suggest a key role for beta5t in generating the MHC class I-restricted CD8(+) T cell repertoire during thymic selection.     

青砖茶不同超滤组分抑制α-淀粉酶和脂肪酶活性研究

结合膜超滤技术从青砖茶中分离提取α-淀粉酶和脂肪酶抑制活性组分,分析青砖茶汤直接超滤组分和青砖茶汤乙酸乙酯萃取物超滤分离组分对α-淀粉酶和脂肪酶活性的影响。结果表明,青砖茶5ku膜截留组分具有较强的α-淀粉酶抑制活性;100ku膜截留组分具有较强脂肪酶抑制活性。乙酸乙酯萃取物5ku膜截留组分具有更强的α-淀粉酶抑制活性,而其透过组分具有更强的脂肪酶抑制活性。膜超滤技术结合乙酸乙酯萃取法能更好地分离纯化青砖茶汤α-淀粉酶和脂肪酶抑制活性成分。     

乳清蛋白抗氧化肽对H_2O_2所致人胚肺成纤维细胞MRC-5过氧化损伤的保护作用

【目的】探讨纯化后的乳清蛋白抗氧化肽P4对人胚肺成纤维细胞(human lung fibroblast)MRC-5过氧化损伤的保护作用及可能的作用机制。【方法】采用H2O2诱导建立细胞氧化损伤模型,应用四唑蓝快速比色法(MTT法)检测细胞存活率,通过检测细胞培养液中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)含量,来确定P4对过氧化损伤MRC-5细胞的保护作用,并利用电镜观察细胞形态学变化。【结果】1mmol·L-1H2O2孵育24h可显著诱导MRC-5细胞损伤,使细胞存活力下降到22.47%,细胞经不同浓度的抗氧化肽P4(4、20、100μg·mL-1)与H2O2共孵育后,特别是100μg·mL-1(高剂量组)抗氧化肽P4可使细胞存活率达到44.77%。同时,提高乳清蛋白抗氧化肽P4的浓度,可促进受损的MRC-5细胞修复,提高了SOD、CAT、GSH-Px酶活性,降低MDA含量。扫描电镜和透射电镜观察结果也表明,一定浓度的乳清抗氧化肽对MRC-5细胞具有保护作用。【结论】乳清抗氧化肽通过拮抗H2O2而对MRC-5的过氧化损伤具有保护作用。     

胸腺肽对幼龄雌性小鼠生殖器官指数及性激素水平的影响

研究了胸腺肽对幼龄雌性小鼠生殖器官指数及性激素水平的影响.以幼龄雌性小鼠为试验动物,连续30d皮下注射不同剂量的胸腺肽,观察各剂量胸腺肽处理对幼龄雌性小鼠卵巢、子宫脏器指数的影响,并检测血清中促卵泡激素(FSH)、雌二醇(E_2)的含量.结果表明,与正常饲养对照组比较,各剂量胸腺肽处理的小鼠卵巢指数、子宫指数降低,差异显著或极显著(p<0.05,p<0.01).其中,0.30mg·mL~(-1)胸腺肽处理组小鼠卵巢指数最低,比正常饲养对照组下降了 36.07%.0.60mg·mL~(-1)胸腺肽组小鼠子宫指数最低,比正常饲养对照组下降了38.08%.与正常饲养对照组相比,各剂量胸腺肽处理小鼠FSH、E2水平降低,差异显著或极显著(p<0.05,p<0.01).其中,0.60mg·mL~(-1)胸腺肽组FSH和E2含量都为最低,分别比正常饲养对照组下降14.48%和17.59%.可见,胸腺肽能降低幼龄雌性小鼠生殖器官指数及血清中性激素水平,从而影响幼龄雌性小鼠的生殖内分泌系统.     

贻贝抗菌肽的分离纯化

[目的]从贻贝中提取具有抗菌作用的多肽。[方法]以紫贻贝为原料,采用0.5%的乙酸直接提取方式提取抗菌肽,再经过Sephacryl S-100聚丙烯酰胺凝胶色谱进行纯化,收集各馏分并用滤纸片扩散法测定其抗菌活性及对各种细菌的最低抑菌浓度,使用SDS-PAGE测定抗菌肽的分子质量,测定其在100℃条件下不同处理时间和不同pH条件下抗菌活性的变化。[结果]使用0.5%的乙酸作为提取液粗提取抗菌肽具有较高的提取效率,使用Sephacryl S-100纯化抗菌肽,可将抗菌肽纯化为单一物质。分离纯化出的贻贝抗菌肽具有较强的抗菌性,分子质量为5 908,且具有耐高温、耐酸碱的性质。[结论]该研究为抗菌肽的大规模生产奠定了基础。