Effect of cortisol on the metabolism of 17-hydroxyprogesterone by Arctic charr and rainbow trout embryos

The effect of cortisol on the in vitro metabolism of [³H]17-hydroxyprogesterone ([³H]17OHP) was studied during embryonic development of Arctic charr (Salvelinus alpinus) and rainbow trout (Oncorhynchus mykiss). In the absence of cortisol, rainbow trout embryos metabolized [³H]17OHP largely to androstenedione (A₄) and androstenetrione (11-KA) with a minor conversion to 17,20ß-dihydroxy-4-pregnen-3-one (17,20P). In the presence of cortisol, this biosynthesis was inhibited. On the other hand, cortisol had no apparent inhibitory effect on the nature of metabolism of [³H]17OHP by Arctic charr embryos. In these embryos [³H]17OHP was metabolized mainly to 17,20P with a minor conversion to A₄ and without the formation of 11-KA that was seen in rainbow trout.When incubated in the presence of [³H]cortisol both Arctic charr and rainbow trout embryos produced 11ß-hydroxyandrostenedione (11ß-OHA) as the major metabolite, with a minor conversion to an unknown steroid. The catabolism of the cortisol by salmonid embryos may reflect the ability of the embryo to inactivate or detoxify cortisol to protect itself from the adverse effects of this biologically potent steroid hormone The study indicates the existence of species-specific differences in the nature of metabolism of [³H]17OHP and the inhibitory effect of cortisol on this metabolism.     

不同发育时期雄性鳗鲡精巢和GtH分泌细胞的结构观察及相关组织中性类固醇激素的变化

通过组织切片和电子显微镜观察经多次注射人绒毛膜促性腺激素(human chorionic gonadotropin,hCG)和鲤鱼脑垂体匀浆液(carp pituitary extract,CPE)促熟的雄性鳗鲡(Anguilla japonica)精巢和脑垂体超微结构的变化,并采用超高效液相色谱-串联质谱(UPLC/MS/MS)联用的方法,观察性腺和血液中7种类固醇激素含量的变化。结果发现:成熟组鳗鲡精巢发育至第Ⅴ期,排精10 d时鳗鲡精巢进入退化吸收期。性成熟雄性鳗鲡的脑垂体GtH分泌细胞代谢活动旺盛,细胞中分泌小球增多,囊泡数量增加,线粒体和内质网丰富。鳗鲡精巢和血液中睾酮(testosterone,T)、17α,20β-二羟基-4-孕烯-3-酮(17α,20β-dihydroxy-4-pregnen-3-one,DHP)和17α-羟孕酮(17α-hydroxyprogesterone,17α-OHP)3种激素含量较高,且成熟的和排精10 d后雄鱼精巢和血液中T、DHP和17α-OHP的含量均显著高于对照组(P﹤0.05),排精10 d时雄鱼精巢和血液中17α-羟孕酮的含量显著低于成熟组鳗鲡精巢和血液中的含量(P﹤0.05);精巢和血液中未检出孕酮(progesterone,P)、雌酮(estrone,E1)、β-雌二醇(estradiol-17β,E2)和雌三醇(estriol,E3)4种激素。结果表明,睾酮、17α-羟孕酮和DHP是雄鱼成熟的关键类固醇激素,其含量在性成熟时达到峰值,排精10 d时含量降低,其含量受脑垂体GtH分泌的调控。     

高密度养殖彭泽鲫造成雄鱼较多的分子机制及成因分析

前期将同一批经雌核发育产生的F1彭泽鲫仔鱼(Pcc)分别置于实验室和池塘进行养殖,发现实验室养殖缸中出现了高比例的雄鱼。本研究为验证雌雄比例是否与密度有关,采用F1彭泽鲫雌鱼经人工雌核发育技术繁育出PccF 2,分高、低密度在实验室进行养殖,并比较了高、低养殖密度下雌雄鱼中性腺分化相关基因的表达。结果表明在PccF 2高、低密度养殖组中,雌鱼中dmrt1c、抗缪勒氏管激素基因amh、芳香化酶基因cyp19a1a的表达显著高于雄鱼中对应基因的表达;雌鱼中dmrt1a、dmrt1b、3β羟基类固醇脱氢酶(3bhsd)、11β羟基类固醇脱氢酶2(11bhsd2)、17α羟化酶/17,20碳链裂解酶a1(cyp17a1)和类固醇激素急性调节蛋白(star)的表达显著低于雄鱼中对应基因的表达。实验室不同密度养殖、不同性别PccF 2彭泽鲫中差异表达基因主要为类固醇合成酶类基因,这些基因的表达差异可能造成雄鱼过多。     

液体悬浮法生产猪苓有效成分的初步研究

以甾体含量为主要指标,以菌丝体生长量及多糖含量为辅助指标,对液体悬浮法生产猪苓甾体所需的培养条件进行了初步研究,结果表明:适合猪苓菌丝甾体化合物积累的最佳液体培养条件为:葡萄糖28.0 g.L-1,酵母膏4.0 g.L-1,磷酸二氢钾1.00 g.L-1,硫酸镁1.00 g.L-1,维生素VB10.1 g.L-1,1%羟甲基纤维素钠10 mL.L-1;在此培养基中每瓶所得菌丝体干重得率为2.050 g,干菌丝体及发酵液中甾体含量分别达到3.864 mg.g-1和37.68 mg.L-1;同时在此培养条件下,菌丝体和发酵液的猪苓多糖含量分别为11.542 mg.g-1和58.85 mg.L-1。     

Bioactive Polyoxygenated Steroids from the South China Sea Soft Coral,Sarcophyton sp

Seven new polyoxygenated steroids (1–7) were isolated together with seven known analogues (8–14) from the South China Sea soft coral, Sarcophyton sp. The structures of the new compounds were identified on the basis of extensive spectroscopic analysis and comparison with reported data. All the steroids are characterized with 3β,5α,6β-hydroxy moiety, displaying carbon skeletons of cholestane, ergostane, gorgostane and 23,24-dimethyl cholestane. In the in vitro bioassay, metabolites exhibited different levels of antimicrobial activity against bacterial species Escherichia coli and Bacillus megaterium, and fungal species Microbotryum violaceum and Septoria tritici. No inhibition was detected towards microalga Chlorella fusca. Preliminary structure-activity analysis suggests that the 11α-acetoxy group may increase both antibacterial and antifungal activities. The terminal-double bond and the cyclopropane moiety at the side chain may also contribute to the bioactivity.     

Differentiation and development of Leydig cell, and induction of spermatogenesis during testicular differentiation in the Japanese eel, Anguilla japonica

The initial appearance and the development of Leydig cells (LCs), the sites of steroid hormone production in the testis, were investigated ultrastructurally during testicular differentiation in the Japanese eel, Anguilla japonica. In addition, the effects of a single injection of human chorionic gonadotropin (HCG; 5 IU g body weight^-1) on histological changes of the testes and serum 11-ketotestosterone (11-KT) were examined at various stages (15–18, 20–23, 26–29, 32–35, 38–41 and 46–50 cm body length (BL)) of testicular differentiation. Testicular differentiation was morphologically characterized by the development of loose connective tissue on the medial side in animals 18–29 cm in BL. Ultrastructurally, LCs were first identified in the loose connective tissue of the testis of the 23 cm fish. In the testes of fish over 32 cm, clusters of LCs were distributed throughout the interstitial region accompanying the increase in number of spermatogonia. In fish larger than 32 cm, spermatogenesis was induced by administration of HCG; serum 11-KT levels were also raised. On the other hand, there was no effect on spermatogenesis or serum 11-KT levels in fish less than 29 cm, or in the controls. These result suggests that morphological differentiation of LCs occurs in testis of the 23 cm eel, and subsequently, the testes of eels of BL more than 32 cm acquire the capability to produce steroid hormones.     

3β,5α,6β-三羟基-16-孕甾烯酮的生物合成及晶体结构分析

利用绿色木霉对5,6α-环氧-3β-羟基-5α-孕甾-16-稀-20-酮进行生物转化得到3β,5α,6β-三羟基孕甾-16-烯-20-酮和3β,5α,6β,15β-四羟基孕甾-16-稀-20-酮。利用IR、MS和2D-NMR实验技术等波谱方法对生物转化产物的结构进行了详细研究,运用X-射线单晶衍射方法确定3β,5α,6β-三羟基孕甾-16-烯-20-酮的空间构型,并进行了晶体结构分析。     

不同温度下雌激素对施氏鲟早期性腺发育及血清性激素的影响

在不同养殖温度下注射外源雌激素后对孵化后90日龄的施氏鲟性未分化的仔鱼进行培育,研究早期性腺发育过程中组织结构和血清性激素含量的变化,以了解温度和外源雌激素对施氏鲟幼鱼性分化和性别比率的影响,探讨施氏鲟的早期性别控制方法和机制。在不同试验温度下,注射外源雌激素-17β-estradiol(E2)30 d后,使施氏鲟幼鱼的卵巢分化提前,但未能改变施氏鲟雌雄1∶1的性别比率。注射24 h后各温度组血浆T含量下降到0 ng.mL-1,显著低于对照组0.46 ng.mL-1的水平(P<0.05)。注射30 d后,血浆T含量有所回升,但以高温组21、24和27℃组的血浆T含量开始回升,27℃组回升幅度最大,与处理前水平无显著差异(P>0.05)。注射后24 h,各温度组血浆E2含量显著升高(P<0.05),7 d后开始缓慢降低,15 d后降到最低值后开始回升,30 d时血浆E2含量与对照组差异不显著(P>0.05)。与注射后血清T含量一样,以温度高组回升快,最快组为21℃组。结果表明,注射17β-E2不能改变施氏鲟的性别比率,但对施氏鲟血浆T和E2含量具有一定影响作用。