灌喂氧化鱼油后黄颡鱼胃肠道黏膜黑色素细胞分化和黑色素合成途径基因的差异表达

以黄颡鱼(Pelteobagrus fulvidraco)为实验对象, 灌喂氧化鱼油、鱼油7d后, 提取胃肠道黏膜总RNA, 采用RNA-seq测序并做转录组分析, 分析了黑色素生物合成途径关键酶(酪氨酸酶)及其相关蛋白基因、黑素体运动的3个蛋白基因、α黑素细胞刺激激素途径和WNT/β-catenin、EDN3和EDNRB、KIT及其配体KITL3个信号通路的主要蛋白基因的差异表达活性。结果显示, 黄颡鱼胃肠道黏膜中存在黑色素细胞分化和发育过程、黑色素合成及其调控途径的代谢网络, 通过绘制代谢网络得到了关键性酶或蛋白质的基因信息。在灌喂氧化鱼油后, 控制黑色素合成途径主要基因的表达活性显著下调, 可能导致黑色素合成量的不足; α-MSH激素途径主要基因差异表达上调, 具备促进黑色素细胞分化和发育的调控基础; 而调控黑色素细胞分化和发育的3个信号通路主要基因也有差异表达。因此, 黄颡鱼受灌喂氧化鱼油的影响, 黑色素细胞分化和发育过程受到较大影响, 会影响到鱼体成熟的黑色素细胞的数量, 同时, 黑色素的生物合成量不足将导致引起黄颡鱼体色的变化。     

黑水虻虫油替代豆油对黄颡鱼幼鱼生长、血清生化指标和肝脏脂滴面积的影响

试验旨在研究黑水虻(Hermetia illucens L.)虫油替代饲料中豆油对黄颡鱼(Pelteobagrus fulvidraco)幼鱼生长性能、血清抗氧化能力、炎症因子及肝脏脂滴面积的影响。以豆油为对照组(T0), 虫油分别替代20%(T20)、40%(T40)、60%(T60)、80%(T80)、100%(T100)的豆油, 配置6种等氮等脂的试验饲料。选取初始体重为(2.12±0.01) g 的黄颡鱼幼鱼840尾, 随机分为6组, 每组4个重复, 每个重复35尾, 养殖56d。结果表明: 黑水虻虫油替代豆油对黄颡鱼幼鱼的末均重、增重率、特定生长率、饲料系数、存活率、摄食率均无显著影响(P>0.05); 各组间黄颡鱼肥满度、腹脂指数、脏体比、肝体比无显著差异(P>0.05); 全鱼粗蛋白、粗脂肪、水分、灰分、钙、磷含量无显著差异(P>0.05); 黑水虻虫油替代豆油对饲料的干物质、蛋白质、脂肪、磷及能量的表观消化率未产生显著影响(P>0.05), 但T60和T80组钙的表观消化率显著高于T100组(P<0.05), 与对照组相比差异不显著(P>0.05)。各组间黄颡鱼幼鱼血清白蛋白、低密度脂蛋白胆固醇、甘油三酯、谷丙转氨酶、谷草转氨酶、尿素氮、球蛋白、乳酸脱氢酶、血糖、总蛋白差异不显著(P>0.05), 但高密度脂蛋白胆固醇含量则随着替代比例的增加先上升后下降, T60组和T80组显著高于对照组(P<0.05)。各组间黄颡鱼幼鱼血清中抗超氧阴离子和丙二醛含量, 过氧化氢酶、超氧化物歧化酶活性无显著差异(P>0.05); 与对照组相比, 各处理组血清肿瘤坏死因子-α浓度无显著性差异(P>0.05); 当黑水虻虫油替代比例超40%时, 各处理组血清中抗炎因子白介素-10和促炎因子白介素-6及白介素-8的浓度均显著高于对照组(P<0.05)。油红O染色显示, 黑水虻虫油替代豆油对肝脏脂滴面积无显著影响(P>0.05)。综上所述, 黑水虻虫油完全替代豆油对黄颡鱼幼鱼生长性能、形体指标、体成分、营养物质表观消化率和抗氧化指标无显著影响, 以生长性能为评价指标, 当黄颡鱼幼鱼基础饲料中添加4%豆油时, 黑水虻虫油完全替代豆油。     

lpa-miR-nov-66通过调控cAMP路径抑制α-MSH介导的羊驼黑色素生成

α-黑素细胞刺激素(α-MSH)和lpa-miR-nov-66在羊驼黑色素细胞产生黑色素过程中均起重要的调控作用,但二者之间的关系尚未报道.本研究在体外培养的羊驼黑色素细胞中通过转染lpamiR-nov-66和添加α-MSH处理,用实时定量PCR和Western印迹检测黑色素细胞内基因表达水平,ELISA法检测c AMP和cGMP的产量,RTCA实时无标记细胞功能分析黑色素细胞增殖以及紫外分光光度法检测黑色素产量,证实二者在调控羊驼黑色素细胞产生黑色素颗粒过程中的关系.结果显示,与单纯α-MSH处理相比,lpa-miR-nov-66转染结合α-MSH处理组中,小眼转录因子(MITF)和酪氨酸酶(TYR)在转录水平和翻译水平的表达均降低,而酪氨酸酶相关蛋白2(TYRP2)在转录和翻译水平的表达均升高;cGMP的产量升高,cAMP的产量下降;黑色素细胞增殖没有显著变化;黑色素细胞内黑色素产量下降.与单纯转染lpa-miR-nov-66相比,lpa-miR-nov-66转染结合α-MSH处理组中,MITF、TYR和TYRP2在转录水平和翻译水平的表达均升高;cGMP的产量下降,cAMP的产量升高;黑色素细胞增殖没有显著变化;黑色素细胞内黑色素产量升高.上述结果证明,lpa-miR-nov-66通过调控羊驼黑色素细胞中毛色形成的c AMP路径,抑制α-MSH对黑色素细胞产生黑色素的促进作用.     

环磷酸腺苷促进泰和乌骨鸡皮肤黑色素细胞黑色素合成

该实验旨在研究环磷酸腺苷(cyclic adenosine monophosphate,c AMP)作为α-黑色素细胞刺激素(α-melanocyte stimulating hormone,α-MSH)-黑素皮质素受体1(melanocortin 1 receptor,MC1R)通路的下游信号分子对泰和乌骨鸡皮肤黑色素细胞黑色素合成的影响。利用体外培养的泰和乌骨鸡皮肤黑色素细胞,观察不同浓度c AMP(0、1×10–5、1×10–4、1×10–3 mol/L)及其抑制剂、腺苷酸环化酶(adenylate cyclase,AC)抑制剂对乌骨鸡皮肤黑色素细胞酪氨酸酶(tyrosinase,TYR)活性和黑色素含量的影响。结果表明,与不添加c AMP的对照组相比,不同浓度的c AMP均极显著提高泰和乌骨鸡皮肤黑色素细胞TYR活性(P0.01),10–4 mol/L组提高的幅度最大。不同浓度的c AMP可不同程度地促进黑色素细胞黑色素的合成,1×10–5 mol/L和1×10–4 mol/L组黑色素含量分别显著(P0.05)和极显著(P0.01)高于不添加c AMP的对照组。c AMP抑制剂Rp-c AMPS预处理黑色素细胞显著抑制c AMP(1×10–4 mol/L)作用下酪氨酸酶活性(P0.01)和黑色素含量的升高(P0.05)。Rpc AMPS和AC抑制剂NKY80预处理黑色素细胞均显著抑制α-MSH(2.5μg/m L)引起的TYR活性、c AMP含量和黑色素含量的升高(P0.01或P0.05)。c AMP作为α-MSH-MC1R信号通路中的第二信使或下游信号分子在泰和乌骨鸡皮肤黑色素细胞黑色素的合成中发挥重要作用,其浓度的升高可提高泰和乌骨鸡皮肤黑色素细胞酪氨酸酶活性以及黑色素的合成。     

饲料中棕榈油替代鱼油和豆油对黄颡鱼生长和肌肉脂肪酸组成的影响

为研究不同棕榈油替代水平对黄颡鱼生长性能、肌肉脂肪酸组成、形体指标和肝脏组织结构的影响, 实验采用5组等氮(400 g/kg)等脂(100 g/kg)等能(15.70 MJ/kg)饲料饲喂黄颡鱼[(16.15±0.04) g], 对照饲料以鱼油鲶豆油1鲶2混合为脂肪源, 实验饲料以棕榈油分别替代10、25、40和55 g/kg的混合脂肪源。经过8周的养殖实验, 结果表明棕榈油替代水平对黄颡鱼的摄食量、终末体重、特定生长率、饲料效率和蛋白沉积率未产生显著性的影响(P>0.05), 但是上述指标均呈现先升高后略微下降的趋势, 并在25 g/kg替代组达到最大值。背肌肌肉中亚油酸和n-3HUFA含量随着棕榈油替代水平的上升而显著下降(P<0.05), 但n-3/n-6比值随着棕榈油替代水平的上升而显著上升(P<0.05), 棕榈油替代水平对各组黄颡鱼的肥满度和脏体比未产生显著性的影响(P>0.05), 但是55 g/kg棕榈油替代水平组肝体比显著高于25 g/kg棕榈油添加组(P<0.05), 且55 g/kg棕榈油替代水平组黄颡鱼的肝脏组织出现细胞肿胀, 细胞核移位, 肝血窦和脂肪细胞数量明显增多等不良影响。据上所述: 对于黄颡鱼幼鱼, 棕榈油可以替代25 g/kg的混合脂肪源(鱼油鲶豆油=1鲶2)不影响鱼体生长性能并且在一定程度上改善了背肌肌肉脂肪酸组成。     

灌喂氧化鱼油对黄颡鱼胃肠道黏膜胆固醇和胆汁酸合成途径基因差异表达的影响

分别提取灌喂氧化鱼油以及鱼油的黄颡鱼胃肠道肠道黏膜的总RNA。将氧化鱼油组、鱼油组总RNA等量混合后,采用RNA-Seq测序,用Trinity进行de novo拼接、组装,对单一基因进行功能注释;再将氧化鱼油组、鱼油组总RNA分别测序、注释后进行基因表达量分析,并计算氧化鱼油组对鱼油组单一基因的差异表达定量分析,以log_2(OFH/FH)值代表氧化鱼油组相对于鱼油组基因的差异表达量。基因差异表达显示,氧化鱼油导致黄颡鱼胃肠道黏膜组织中胆固醇、胆汁酸生物合成代谢途径的酶以及涉及胆固醇和胆汁酸吸收转运蛋白基因差异表达。黄颡鱼胃肠道黏膜中以乙酰辅酶A为原料的胆固醇生物合成途径关键酶基因差异表达显著下调,从细胞外吸收转运胆固醇的主要蛋白基因差异表达下调,显示灌喂氧化鱼油导致黄颡鱼胃肠道黏膜胆固醇合成能力下降、从其他组织吸收胆固醇的能力下降,血清胆固醇含量下降4.06%。灌喂氧化鱼油后,黄颡鱼胃肠道黏膜以胆固醇为原料的初级胆汁酸合成代谢的关键酶差异表达显著上调,而胆汁酸转运到细胞外和转运到肠腔的载体蛋白基因差异表达下调,肠道再吸收胆汁酸的转运载体蛋白基因差异表达下调及肝细胞从血液吸收转运的载体蛋白基因差异表达上调,而血清胆汁酸含量下降了20.00%。结果表明,胆汁酸的肠肝循环发生障碍,肠腔和血清胆汁酸含量下降、胆汁酸在细胞出现淤积的趋势。     

饲料中添加螺旋藻和叶黄素对杂交黄颡鱼生长、抗氧化能力和体色异常调控的比较研究

研究以杂交黄颡鱼(黄颡鱼Pelteobagrus fulvidraco♀×瓦氏黄颡鱼Pelteobaggrus vachelli♂)“黄优一号”[体重(4.79±0.01) g]为研究对象, 探究饲料中螺旋藻(Arthrospira platensis)和叶黄素对黄颡鱼生长、抗氧化能力和抗病能力及体色异常调控的影响。实验设计4组饲料, 不添加任何色素源的对照组(Con), 添加2%的螺旋藻组(Spirulina, SP), 添加0.2%的叶黄素组(Lutein, Lut)及添加2%螺旋藻和0.1%溶血磷脂组(Spirulina+Lysophospholipids SPL)。720尾鱼随机放入12个缸, 每缸60尾鱼, 先使用对照组饲料投喂20d至体表颜色异常, 再用4种饲料饲养35d, 共4个处理, 每个处理3个重复。在养殖实验结束后进行嗜水气单胞菌攻毒实验。研究结果表明, 各个处理组间特定生长率(SGR)和摄食率(FR)均没有显著差异(P>0.05); 但SP和SPL组饲料效率(FE)显著高于对照组(P<0.05)。各组间全鱼水分、 蛋白和脂肪含量没有显著差异(P>0.05)。各组间实验鱼皮肤亮度没有显著性差异(P>0.05); 但SP和SPL组皮肤黄度和饱和度显著高于另外两组(P<0.05); 同时, SP和SPL组背部皮肤红度显著高于对照组(P<0.05); SPL组腹部皮肤红度显著高于对照组(P<0.05)。SP和SPL组腹部皮肤叶黄素含量显著高于对照组(P<0.05)。对照组超氧化物歧化酶(SOD)活性显著低于另外3个处理组(P<0.05); 各组过氧化氢酶(CAT)活性没有显著性差异(P>0.05); SPL组谷胱甘肽过氧化物酶(GPX)活性和还原型谷胱甘肽(GSH)含量显著高于叶黄素和对照组(P<0.05); SP和SPL组丙二醛(MDA)含量显著低于另外2个处理组(P<0.05)。SP和SPL组的实验鱼感染嗜水气单胞菌后的累计存活率显著高于叶黄素和对照组(P<0.05)。综上所述, 黄颡鱼“黄优一号”在前期投喂缺乏色素的饲料造成体色异常后, 在饲料中添加螺旋藻或配合添加溶血卵磷脂比叶黄素更有效的改善鱼体的体色, 同时更能提高黄颡鱼的抗氧化能力和抗病力。     

蝇蛆粉替代鱼粉对杂交黄颡鱼生长性能、体组成和抗氧化能力的影响

研究旨在评估蝇蛆粉(MGM)替代鱼粉(FM)对杂交黄颡鱼(Pelteobagrus fulvidraco)"黄优1号"幼鱼生长、体组成和抗氧化能力的影响。实验设计6组等氮(粗蛋白42%)饲料,分别使用蝇蛆粉替代0、20%、40%、60%、80%和100%的鱼粉蛋白,分别记为G0、G20、G40、G60、G80和G100。将初始体重为(0.62±0.01) g的杂交黄颡鱼幼鱼随机分为6组,进行为期8周的投喂实验。实验结束时测定其生长性能、全鱼体组成和肝组织与血清中的抗氧化酶活性(SOD、CAT、GR和T-AOC)。结果显示:(1)与G0相比, G20与G40组杂交黄颡鱼幼鱼的特定生长率并未出现显著性差异(P>0.05), G60、G80与G100组的幼鱼的特定生长率表现出显著的负面影响(P<0.05)。与G0相比, G20与G40组的饲料系数并未出现显著性差异(P>0.05),G60、G80与G100的饲料系数显著性升高(P<0.05)。不同替代组杂交黄颡鱼幼鱼的存活率没有显著差异(P>0.05)。(2)各替代组杂交黄颡鱼幼鱼的全鱼体组成与形体指标没有显著差...     

凡纳滨对虾饲料中酵母水解物替代鱼粉适宜比例的研究

试验添加酵母水解物替代不同比例鱼粉, 添加量分别为0(对照组)、2.5%、5.0%、7.5%、10%和12.5%,替代鱼粉的比例分别为0、8%、16%、24%、32%和40%, 通过评估生长性能、消化酶和免疫相关酶活性等指标评价酵母水解物替代鱼粉对凡纳滨对虾健康生长的影响。选择健康凡纳滨对虾[初重(0.630.01) g]随机分为6组, 每组3个重复, 养殖8周。结果表明, Y2.5和Y5.0处理组对虾增重率和特定生长率与Y0相比差异不显著(P0.05), 但是显著高于其余各组(P0.05), Y2.5组饲料系数和Y0组相比差异不显著, 显著低于其余各组(P0.05); Y5.0组肝胰腺糜蛋白酶和Y0组相比差异不显著(P0.05), 显著高于其余各组(P0.05); Y2.5和Y7.5组胰蛋白酶和Y0组相比差异不显著, 但是显著高于其余各组(P0.05); Y2.5组与Y5.0酚氧化酶活性显著高于其余各组(P0.05), Y2.5组与Y7.5组和Y0组总一氧化氮合酶活性显著高于其余各组(P0.05), Y5.0组和Y7.5组血清溶菌酶活性显著高于其余各组(P0.05)。以增重率为判据, 经二次曲线拟合得出, 获得最大增重率时酵母水解物添加量为1.62%, 替代鱼粉比例为5.19%; 酵母水解物替代鱼粉比例达24%时不会对增重率产生显著影响。     

开放流水养殖模式下暂养时间对黄颡鱼肌肉品质和营养价值的影响

文章旨在探索开放流水养殖模式下,暂养对黄颡鱼(Pelteobagrus fulvidraco)肌肉品质和营养价值的影响。试验选用处于快速生长期的大规格鱼种(15.69±2.28) g黄颡鱼为试验对象,随机分为4组,分别暂养0(对照组)、20d、30d和40d,每组3个重复。试验期间投喂黄颡鱼配合饲料,在养殖过程中定期测量水质。在试验结束后对其生长性能、血清生化指标、肌肉氨基酸、脂肪酸组成及质构特性进行比较。结果显示:(1)暂养期间试验池的水质氨态氮含量在0.03—0.05 mg/L,亚硝态氮含量均在0.01 mg/L,溶氧水平均超过9.0 mg/L。(2)暂养时间延长,黄颡鱼末体重和增重率呈上升趋势,特定生长率呈下降趋势,各组间差异显著(P<0.05);暂养20d、30d和40d组的成活率、肥满度、肝体比及脏体比组间无显著性差异(P>0.05)。(3)暂养30d和40d组血清中的白蛋白和总蛋白含量显著高于对照组和暂养20d组(P<0.05),但其γ-谷氨酰基转移酶和碱性磷酸酶活性、总胆固醇和总胆汁酸含量显著低于对照组和暂养20d组(P<0.05);对照组、暂养2...     

Cultured Human Melanocytes Respond to MSH Peptides and ACTH

Although the administration of melanocyte-stimulating hormone (MSH) peptides results in skin darkening in man, cultured human melanocytes have been reported to be unresponsive to these peptides. This may be a consequence of the conditions under which the cells were maintained in vitro, particularly the use of phorbol esters and cholera toxin as melanocyte mitogens. By culturing the cells in the absence of these additives, we demonstrate that α-MSH and its synthetic analogue Nle⁴DPhe⁷α-MSH (NDP-MSH) induce dose-related increases in melanin content and tyrosinase activity and affect cell morphology in the majority of human melanocyte cultures. In addition, NDP-MSH induces increases in tyrosinase mRNA and tyrosinase-related protein-1 (TRP-1) mRNA. The dose-response curves for the MSH peptides are sigmoidal and the two peptides are equipotent in their effects on human melanocytes. Adrenocorticotropic hormone (ACTH) also affects morphology and stimulates melanogenesis and tyrosinase activity in human melanocytes. However, the dose-response curves for ACTH are biphasic, and the melanocytes respond to lower concentrations of ACTH than MSH peptides, similar to those normally present in human plasma. These findings may be important in understanding the role of these pro-opiomelanocortin peptides in human skin pigmentation.     

亚麻油替代鱼油对杂交鲟生长、脂肪酸组成及脂肪代谢的影响

为研究亚麻油替代不同水平的鱼油后对杂交鲟(Acipenser baeri Brandt♀×A. schrenckii Brandt♂)幼鱼[初均重(70.8±0.5) g]生长、脂肪酸组成、肝脏及肌肉脂肪沉积以及脂肪代谢的影响, 在油脂添加量为8%的饲料中用亚麻油分别替代0(LO0)、25%(LO25)、50%(LO50)、75%(LO75)和100%(LO100)的鱼油, 配制5种等氮(38.7%CP)等脂(10%CF)饲料。每组饲料随机设3个重复, 养殖周期为12周。结果表明,亚麻油替代100%的鱼油对杂交鲟幼鱼的生长没有显著影响, 而且随着饲料中亚麻油含量的上升, 饲料效率有所提高, 100%鱼油替代组的饲料效率明显高于100%鱼油组的(P<0.05); 但用亚麻油替代鱼油后, 肌肉和肝脏的粗脂肪含量以及血清中谷草转氨酶、谷丙转氨酶和乳酸脱氢酶活性明显升高(P<0.05); 肌肉亚麻酸和n-3多不饱和脂肪酸的含量与饲料中相应脂肪酸组成呈明显的线性相关关系(R2>0.69; P<0.05)。对于杂交鲟的脂肪代谢而言, 亚麻油的添加对血清中的游离脂肪酸、甘油三酯、高、低密度脂蛋白胆固醇的变化产生明显影响, 但亚麻油对血清总胆固醇和酮体影响不显著。考虑到亚麻油完全替代鱼油后, 肌肉中的EPA和DHA这两种长链高不饱和脂肪酸的含量仅下降了不到30%, 因此亚麻油应该是一种比较优质的鱼油替代品。     

ANTAGONIST AND AGONIST ACTIVITIES OF THE MOUSE AGOUTI PROTEIN FRAGMENT (91–131) AT THE MELANOCORTIN-1 RECEPTOR

Antagonist and agonist activities of chemically synthetized mouse agouti protein fragment (91–131) (AP_91–131) at the melanocortin type-1 receptor (MC1-R) were assessed using B16-F1 mouse melanoma cells in vitro and the following assay systems: (i) receptor binding, (ii) adenylate cyclase, (iii) tyrosinase, (iv) melanin production, and (v) cell proliferation. In competition binding studies AP_91–131 was about 3-fold less potent than the natural agonist α-melanocyte-stimulating hormone (α-MSH) in displacing the radioligand [¹²⁵I]-[Nle⁴, D-Phe⁷]-α-MSH (K_i 6.5±0.8 nmol/l). α-MSH-induced tyrosinase activation and melanin production were completely inhibited by a 100-fold higher concentration of AP_91–131; the IC₅₀ values for AP_91–131 in the two assay systems were 91±22 nM and 95±15 nM respectively. Basal melanin production and adenylate cyclase activity in the absence of agonist were decreased by AP_91–131 with IC₅₀ values of 9.6±1.8 nM and 5.0±2.4 nM, respectively. This indicates inverse agonist activity of AP_91–131 similar to that of native AP. The presence of 10 nM melanin-concentrating hormone (MCH) slightly potentiated the inhibitory activity of AP_91–131 in the adenylate cyclase and melanin assays. On the other hand, AP_91–131 inhibited cell growth similar to α-MSH (IC₅₀ 11.0±2.1 nM; maximal inhibition 1.8-fold higher than that of α-MSH). Furthermore, MC1-R was down-regulated by AP_91–131 with about the same potency and time-course as with α-MSH. These results demonstrate that AP_91–131 displays both agonist and antagonist activities at the MC1-R and hence that it is the cysteine-rich region of agouti protein which inhibits and mimics the different α-MSH functions, most likely by simultaneous modulation of different intracellular signalling pathways.     

Analysis of the UV-Induced Melanogenesis and Growth Arrest of Human Melanocytes

Cultured human melanocytes derived from different skin types responded to frequent treatment with ultraviolet (UV) light with increased melanin synthesis, decreased proliferation, and morphologic signs of aging. These effects were augmented by increased frequency of irradiation with 15.5 mJ/cm² UV light. Stimulation of melanogenesis by UV light involved an increase in tyrosinase activity, without any change in the amounts of either tyrosinase or tyrosinase-related protein (TRP)-1, and a decrease in the amount of TRP-2, as determined by Western blot analysis. These results are different from the mechanisms by which other melanogenic agents, such as cholera toxin and isobutyl methylxanthine, stimulated melanogenesis, whereby the amounts of tyrosinase, TRP-1 and TRP-2 were increased. The decrease in the amount of TRP-2 might be significant in that it might alter the properties of the newly synthesized melanin. The UV irradiation protocol that was followed blocked melanocytes in G₂-M phase of the cell cycle without compromising cellular viability. Following three rounds of UV irradiation, melanocytes could recover from the growth arrest and resume proliferation. Treatment with 0.1 μM α-melanocyte stimulating hormone (α-MSH) postirradiation enhanced the melanogenic effect of UV light and stimulated the melanocytes to proliferate. The effects of α-MSH on the UV induced responses and their implications on photocarcinogenesis are being further investigated. Analyzing the mechanisms by which UV light exposure affects normal melanocytes might lead to a better understanding of how these cells undergo malignant transformation, and why individuals with different skin types differ in their susceptibility to skin cancers.     

氧化油脂对斑点叉尾鮰生长、体色和抗氧化能力的影响

为考察正常及氧化鱼油、豆油、混合油脂(鱼油﹕豆油=1﹕1)对斑点叉尾鮰(Ictalurus punctatus)生长性能、体色和肝脏抗氧化能力的影响, 设计6组等氮等脂饲料, 分别添加6%鱼油、6%豆油、3%鱼油+3%豆油、6%氧化鱼油、6%氧化豆油、3%氧化鱼油+3%氧化豆油, 饲喂6组初始体重(150.5±4.2 g)的斑点叉尾鮰8周, 每组3个重复, 每个重复14尾鱼。结果表明, 摄食6%鱼油、6%豆油、3%鱼油+3%豆油饲料的斑点叉尾鮰在增重率、饲料系数、肌肉组成、体色和肝脏抗氧化指标上均无显著差异(P>0.05); 摄食3组氧化油脂饲料后, 鱼体增重率和肌肉粗脂肪含量降低(P<0.05), 饲料系数和背部、尾部皮肤b*(黄度值)增加(P<0.05), 肝脏丙二醛(MDA)和血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)和直接胆红素升高(DBIL)(P<0.05), 肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、还原型谷胱甘肽降低(GSH)(P<0.05), 而肌肉色度值(L*、a* 、b*)和肌肉水分、粗蛋白质、灰分含量无显著差异(P>0.05)。以上结果表明, 6%氧化油脂(鱼油、豆油或混合油脂)导致斑点叉尾鮰生长性能下降、皮肤黄度增加、肝脏抗氧化能力受损; 豆油可替代斑点叉尾鮰饲料中鱼油的使用, 而不会对生长产生负面影响。     

A novel synthetic compound, (Z)-5-(3-hydroxy-4-methoxybenzylidene)-2-iminothiazolidin-4-one (MHY773) inhibits mushroom tyrosinase

As part of continued efforts for the development of new tyrosinase inhibitors, (Z)-5-(substituted benzylidene)-2-iminothiazolidin-4-one derivatives (1a – 1l) were rationally synthesized and evaluated for their inhibitory potential in vitro. These compounds were designed and synthesized based on the structural attributes of a β-phenyl-α,β-unsaturated carbonyl scaffold template. Among these compounds, (Z)-5-(3-hydroxy-4-methoxybenzylidene)-2-iminothiazolidin-4-one (1e, MHY773) exhibited the greatest tyrosinase inhibition (IC₅₀ = 2.87 μM and 8.06 μM for monophenolase and diphenolase), and outperformed the positive control, kojic acid (IC₅₀ = 15.59 and 31.61 μM). The kinetic and docking studies demonstrated that MHY773 interacted with active site of tyrosinase. Moreover, a melanin quantification assay demonstrated that MHY773 attenuates α-melanocyte-stimulating hormone (α-MSH) and 3-isobutyl-1-methylxanthine (IBMX)-induced melanin contents in B16F10 melanoma cells. Taken together, these data suggest that MHY773 suppressed the melanin production via the inhibition of tyrosinase activity. MHY773 is a promising for the development of effective pharmacological and cosmetic agents for skin-whitening.     

In situ melanin assay for MSH using mouse B16 melanoma cells in culture

A sensitive in situ melanin assay using cultured mouse B16 melanoma cells is described for structure-activity studies with melanocyte-stimulating hormone (MSH) peptides. B16 Cells were seeded at a density of 2500 cells per well in 96-well microtest tissue culture plates; after 24 h the cells were incubated in the presence of serial dilutions of MSH peptides for 3 to 5 days. The melanin released into the medium of each well was then determined spectrophotometrically at a wavelength of 405 nm using an automatic microplate reader calibrated against synthetic melanin. Studies with alpha-MSH, [Nle4, D-Phe7]-alpha-MSH, [3'-iodo-Tyr2]-alpha-MSH, adrenocorticotropin (ACTH)(1-24), and ACTH(1-39) showed that the peptides had identical intrinsic activities and that the relative potencies were similar to those obtained with a tyrosinase assay. The EC50 of alpha-MSH was 27 pM, i.e., about five- to sevenfold lower than that in the assays for tyrosinase or intracellular melanin. Thus, the new assay represents the most sensitive melanoma cell assay for MSH available to date.