Eisenia bicyclis的克隆苗繁育及养殖技术研究

以从日本引进的Eisenia bicyclis雌、雄配子体为材料,研究了不同温度（10、15、20℃）和光照强度[20、40、60μmol/（m2·s）]对雌、雄配子体生长发育和幼孢子体生长的影响。结果表明：雌、雄配子体在温度为20℃时生长最快,15℃时次之,10℃时最慢,在温度为10℃时随光照强度的升高其生长速度加快,但随温度的升高不同光照组间的生长差异逐渐缩小;15℃和20℃是雌、雄配子体发育成熟的适宜温度,培养12 d后配子体的成熟率可达到95%~100%,而且高光照条件下的成熟率明显优于低光照下;幼孢子体生长的适宜温度为20℃,在各光照条件下的生长速度明显快于温度为15、10℃组;温度为10℃时,光照强度越高幼孢子体生长越快,温度为15、20℃时,幼孢子体在光照强度为20μmol/（m2·s）时的生长速度明显慢于40、60μmol/（m2·s）组,而40μmol/（m2·s）组与60μmol/（m2·s）组间生长差异不明显;将雌、雄配子体打碎进行克隆苗繁育,在室内培养至体长为800μm以上的幼孢子体,将其移至海区暂养,1个月后可生长成2 cm左右的幼体。

Evaluation for Antioxidative Properties of Phlorotannins Isolated from the Brown Alga <i>Eisenia bicyclis</i>, by the H-ORAC Method

The antioxidative properties of phlorotannins isolated from the brown alga Eisenia bicyclis were measured using the H-ORAC (Hydrophilic Oxygen Radical Absorbance Capacity) method. The ORAC values of phloroglucinol and it’s oligomers: eckol, fucofuroeckol A, phlorofucofuroeckol A, dieckol, 8,8’-bieckol, were 2.57 ± 0.14, 4.97 ± 0.36, 9.82 ± 0.70, 8.97 ± 0.89, 10.22 ± 0.85, 8.62 ± 0.92 μmol Trolox equivalent/μmol, respectively. With the exception of eckol, the ORAC values of tested phlorotannins were higher than those of the well-known antioxidants (epigallocatechin gallate, resveratrol and L-ascorbic acid) used as positive controls. As a result of comparing with known ORAC values, it was found that the dieckol and fucofuroeckol A had stronger antioxidant activity than representative polyphenols (e.g., kaempferol, quercetin, myricetin and chlorogenic acid) derived from terrestrial plants.

Use of a Natural Compound Made of Ecklonia bicyclis Seaweed, Tribulus terrestris and Water-Soluble Chitosan Oligosaccharide, in Male Sexual Asthenia with Mild or Mild-Moderate Erectile Dysfunction and Serum Testosterone Levels at the Lower Limit of Normal

Objectives: to evaluate the effectiveness of a natural compound made of Ecklonia bicyclis Seaweed, Tribulus terrestris and water-soluble chitosan oligosaccharide, in the male sexual asthenia with mild or mild-moderate erectile dysfunction and serum testosterone levels between 280 and 350 ng/dl. Materials and Methods: 84 male patients affected by reduced libido and serum testosterone levels at the lower limit of normal, were recruited. We have separated patients in three different age groups: group A (18 - 45 years), group B (45 - 59 years), group C (>60 years). All subjects answered the International index of erectile function questionnaire (IIEF-5) and underwent determination of serum total testosterone before and after 30 days of treatment. Results: Before treatment, the group A showed mean (± standard deviation) total testosterone 321.9 ± 19.2 ng/dl and mean IIEF-5 18.6 ± 1.97, in the group B it was 318.5 ± 18.1 ng/dl and 16.3 ± 2.66, and finally in the group C it was 305.4 ± 13.1 ng/dl and 14.2 ± 1.95 respectively. After treatment mean total testosterone and mean IIEF-5 were respectively: group A (448 ± 111.46 ng/dl and 21.84 ± 3.41);group B (453.8 ± 105.23 ng/dl and 20.4 ± 3.81);group C (385.8 ± 87.29 ng/dl and 16.7 ± 3.84). Conclusions: The treatment with Ecklonia bicyclis, Tribulus terrestris and water-soluble chitosan oligosaccharide might represent a safe and effective option on the improvement of libido and erectile function in man with testosterone level at the lower limit of normal.

Physiological differences in the growth and maturation of Eisenia bicyclis and Ecklonia cava gametophytes in Korea

The objective of this study was to examine the eff ects of temperature and light intensity on growth of female gametophytes of Eisenia bicyclis and Ecklonia cava and responses of these female gametophytes to Fe addition and daylength. Female gametophytes of each species were cultured at four temperatures (10, 15, 20, and 25°C) and under a combination of four light intensities (10, 20, 40, and 80 μmol photons/(m^2 ·s)) with two temperatures (15 and 20°C for E i . bicyclis;20 and 25°C for E c . cava ) to clarify their optimal growth conditions. Growth and maturation of female gametophytes of these two species under a combination of five Fe-EDTA concentrations (0, 1, 2, 4, and 8 μmol/L) and three daylengths (10, 12, and 14 h) were also examined. The growth of E i . bicyclis gametophyte was maximal at approximately 15–20°C, 20 μmol photons/(m^2 ·s), Fe-EDTA concentration of 8 μmol/L and daylengths of 12–14 h. While E c . c ava gametophytes showed optimal growth at approximately 20–25°C, 20 μmol photons/(m^2 ·s), Fe- EDTA concentration of 8 μmol/L and daylength of 14 h. Maturation of gametophytes was enhanced at Fe-EDTA concentration of 4 μmol/L for E i . bicyclis and at 2 μmol/L for E c . c ava . In conclusion, optimal growth temperatures and Fe-EDTA concentrations for maturation of E i . bicyclis and E c . c ava gametophytes were diff erent. Higher optimal growth temperature for E c . c ava gametophytes may contribute to its wider geographical distribution compared to E i . bicyclis which has restricted habitats in Korea. This suggests that addition of Fe ion could be used to recover beds of these two species in barren grounds of Korea.

Dieckol isolated from Eisenia bicyclis extract suppresses RANKL-induced osteoclastogenesis in murine RAW 264.7 cells

Objective:To demonstrate the effect of dieckol from Eisenia bicyclis on osteoclastogenesis using RAW 264.7 cells.Methods:Murine macrophage RAW 264.7 cells were subjected to dieckol treatment,followed by treatment with receptor activator of nuclear factor kappa-B ligand(RANKL)to induce osteoclastogenesis.Tartrate-resistant acid phosphatase(TRAP)activity was examined using a TRAP activity kit.Western blotting analysis was conducted to examine the level of osteoclast-related factors,including TRAP and calcitonin receptor(CTR),transcriptional factors,including c-Fos,c-Jun,and nuclear factor of activated T cells cytoplasmic 1(NFATc1),nuclear factor kappa-B(NF-κB),extracellular signal-regulated kinase(ERK),and c-Jun N-terminal kinase(JNK).Immunofluorescence staining was conducted to examine the expression of c-Fos,c-Jun,and NFATc1.Results:Among the four phlorotannin compounds present in Eisenia bicyclis,dieckol significantly hindered osteoclast differentiation and expression of RANKL-induced TRAP and CTR.In addition,dieckol downregulated the expression levels of c-Fos,c-Jun,NFATc1,ERK,and JNK,and suppressed NF-κB signaling.Conclusions:Dieckol can suppress RANKL-induced osteoclastogenesis.Therefore,it has therapeutic potential in treating osteoclastogenesis-associated diseases.

Elastosonographic Changes in Patients with Peyronie's Disease, before and after Treatment with a Compound Based on <i>Ecklonia bicyclis</i>, <i>Tribulus terrestris</i>, and Water-Soluble Chitosan

Several diagnostic techniques have been proposed during the time for the diagnostic evaluation of Induratio Penis Plastica (IPP), due to poor sensitivity and specificity of standard B-Mode Ultrasonography. Shear-Wave elastosonography (SWE) is a relative new ultrasound technique, already used for several organs, which allows evaluating the rigidity of the tissues. Furthermore, we used a compound consisting of Ecklonia bicyclis, Tribulus terrestris, and Biovis (water-soluble Chitosan) with antifibrotic and antioxidant properties and analyzed the elastosonographic variations following the assumption of the compound. From March 2016 to April 2017, 40 patients were recruited with a mean age of 62. All patients underwent elastosonographic measurements at the start of treatment. Subsequently each patient took 1 tablet of the compound of Ecklonia bicyclis, Tribulus terrestris, and Biovis (water-soluble Chitosan) for 6 months. All patients underwent elastosonographic investigation at the end of treatment. The elastosonographic measurement prior treatment, showed that the average of the three measurements (proximal, medial and distal), was 37.05 KPa ± 7.76 SD and 38.8 KPa ± 10.11 SD for the left and right cavernous body, respectively. The examination at the end of the six months of treatment, with the compound showed that the average values of fibrosity were 31.07 KPa ± 7.5 SD and 30.86 KPa ± 11.4 SD for the left and right cavernous body respectively, demonstrating a decrease in fibrosity that was statistically significant (P < 0.01). Our data demonstrate that taking the compound for six months reduces the stiffness of the corpora cavernosa.

The Influence of Phlorotannins and Bromophenols on the Feeding Behavior of Marine Herbivorous Gastropod <i>Turbo cornutus</i>

The influence of phlorotannins and bromophenols on the feeding behavior of marine herbivores was determined using bioassay of Turbo cornutus. It was found that phloroglucinol and its oligomers isolated from the brown alga Eisenia bicyclis (eckol, fucofuroeckol A, phlorofucofuroeckol A, dieckol, and 8,8’-bieckol) had a deterrent effect against feeding behavior of T. cornutus in the concentration of algal body, respectively. In the case of the examination of 0.1 mM concentration, although fucofuroeckol A and phlorofucofuroeckol A significantly reduced feeding by T. cornutus, phloroglucinol and 8,8’-bieckol did not show any significant influence on feeding behavior, and eckol and dieckol had stimulating activity. 2,4-Dibromophenol and 2,4,6-tribromophenol, which are major components of extracellular secretions from Eisenia and Ecklonia species, caused the death, the torpidity, and the decreased appetite of T. cornutus at the concentration of 0.1 mM. In addition, 2,4-dibromophenol had strong feeding deterrent activity at the concentration of 1 μM. These results indicate that phlorotannins and bromophenols act as chemical defense agents of brown algae against environmental stresses such as the herbivore attack.

“Tradamixina TP” Compound in Monotherapy for Three Months in Patients with LUTS: Observational Study on Improvement of Urinary Symptoms and Sexual Function in Men

Introduction: Benign Prostatic Hypertrophy (BPH) and Lower Urinary Tract Symptoms (LUTS) frequently affect the male population after the age of 50. Moreover, it is well known that there is a correlation of these symptoms with Erectile Dysfunction (ED) that can reach a frequency of roughly 52% in men aged between 40 and 70 years old. In this study, we aimed at evaluating the effect of a new natural compound “Tradamixina TP” (composed of 80 mg of the algae Ecklonia Bicyclis, 100 mg of Tribulus terrestris and 100 mg of Chitosan Water-soluble oligosaccharide 320 mg of Serenoa repens) in the improvement of Lower Urinary Tract Symptoms (LUTS). Materials & Methods: One-hundred patients with an average age of 64 years old and who were not taking any medication or phytotherapy for prostatic hypertrophy or erectile dysfunction were recruited in the study. All participants were subjected to serum prostate antigen (PSA) and uroflowmetry measuring maximal urine flow/sec (Qmax), at baseline and after 12 weeks. In addition, patients were asked to take the Tradamixina TP for 3 months and to complete at the beginning and end of the treatment the following questionnaires: 1) the IPSS investigating the level of LUTS, and 2) the IIEF-15 investigating the erectile dysfunction. Results: After the treatment, the IPSS scores were lower than those before the treatment. Moreover, a significant effect was found for the IIEF erectile subdomain with scores that were significantly higher at 12 weeks than at baseline. A similar result was found for the IIEF overall subdomain. A decrease of total serum PSA was also observed. Conclusions: After 3 months of daily treatment, the new compound “Tradamixina TP” improved patients’ sexual function, LUTS and led also to a decrease in serum PSA levels.