Eco-friendly Catalytic Conversion of Fructose into 5-Hydroxymethylfurfural and 2,5-Diformylfuran : 과당으로부터 5-Hydroxymethylfurfural 및 2,5-Diformylfuran 전환을 위한 친환경적 촉매반응

네하 미뜰 명지대학교 대학원 2015 국내박사

Catalytic conversions of fructose to platform chemicals such as 5-hydroxymethylfurfural (5-HMF) and its furanic derivative, 2,5-diformylfuran (2,5-DFF), were investigated under mild conditions using homogeneous and heterogeneous catalysts. The effects of various factors in the catalytic reactions were evaluated. Mild and homogeneous dehydration systems were examined for the conversion of fructose in the presence of acid catalysts such as phosphorous pentoxide (P2O5) and earth abundant metal chlorides in the ionic liquids. The acidic nature of P2O5 along with its hygroscopic properties has been successfully utilized to afford 81.2 % yield of 5-HMF at 50 oC in 60 min. Different metal chlorides were examined as Lewis acid catalysts for production of 5-HMF from fructose. Among tested metal chlorides, niobium pentachloride (NbCl5) exhibited the highest 5-HMF yield. NbCl5 (0.20 mmol) afforded 79.3 % yield of 5-HMF at 80 oC only within 30 min. It was found that no degradation products were generated under these conditions resulting, while high selectivity for the target product was achieved. FT-IR studies indicated that the moderate Lewis acidity of NbCl5 was possibly responsible for the selective formation of 5-HMF. Besides, an aerobic oxidation system was investigated for derivatization of 5-HMF to 2,5-DFF using 4-hydroxy-2,2,6,6-tetramethyl-piperidine-1-oxyl (4-hydroxy-TEMPO) radical as a catalyst with [bis(acetoxy)-iodo]benzene (BAIB) and acetic acid (CH3COOH) as co-oxidants. The highest yield (65.0 %) of 2,5-DFF was achieved in the optimized TEMPO/BAIB/CH3COOH system at 30 oC within 45 min in ethyl acetate. It was found that the presence of BAIB facilitated in-situ regeneration of hydroxylamine form of TEMPO and acetic acid addition assisted in initial formation of oxoammonium form of TEMPO, which was responsible for facile 5-HMF oxidation. It is projected that the proposed reaction system in this study is environmentally benign (metal-free) and highly energy efficient. In order to recycle TEMPO catalyst effectively, immobilization of 4-hydroxy-TEMPO on SBA-15 was carried out and then, TEMPO-SBA-15 was characterized by XRD, N2 physisorption, thermo-gravimetric analyses (TGA), SEM and FT-IR. TEMPO-SBA-15 efficiently oxidized 5-HMF to 2,5-DFF with 72.5 % yield at only 40 oC in ethyl acetate with the aid of BAIB and acetic acid as co-oxidants in an oxygen-rich environment. It indicated that the pore size distribution and architecture of SBA-15 played important roles for the diffusive transport of reactants and products, which resulted in improved selectivity. In addition, the catalyst was reused for five times with negligible decrease in terms of its catalytic activity. A novel bi-functional catalyst (WO3HO-VO(Salten)-SiO2@Fe3O4) with silica encapsulated magnetic nanoparticles was prepared by anchoring oxovanadium complex and tungstic acid onto the surface of silica coated Fe3O4 nanoparticles. The prepared catalyst and its precursors were characterized by FT-IR, XRD, HRTEM and N2 physisorption, and (TGA). The WO3HO-VO(Salten)-SiO2@Fe3O4 catalyst facilitated a remarkable activity for the one-pot; i.e. direct conversion of fructose to 2,5-DFF in one step. In this manner, tedious separation and purification steps of intermediate compound 5-HMF could be eliminated. Conversion yield (70.4 %) of 2,5-DFF was obtained in an one-pot reaction of fructose at mild reaction temperature of 50 oC. After a reaction, catalyst can simply be recovered by using an external magnet. 균일 및 불균일 촉매군을 사용하여 탄수화물 과당으로부터 플랫폼 화학물질인 5-Hydroxymethylfurfural (5-HMF) 및 퓨란 유도체인 2,5-Diformylfuran (2,5-DFF) 등으로의 화학전환반응에 관한 연구를 수행하면서 다양한 반응조건 및 촉매가 반응에 미치는 영향을 조사하였다. 이온성 액체상에서 P2O5 및 금속염화물과 같은 균일계 산촉매를 사용함으로써 온화한 조건에서 과당의 탈수반응을 연구하였다. P2O5의 흡습성 및 산특성을 활용하여 50 oC에서 60분 반응 시 81.2 % 의 5-HMF 수율을 얻었다. 또한, 다양한 금속염화물을 루이스산 촉매로서 적용해 본 결과 오염화니오븀 (NbCl5)이 과당으로부터 5-HMF 합성에 있어서 최고 수율을 나타내었다. 80 oC에서 NbCl5 (0.20 mmol)을 사용한 결과 단 30분 만에 79.3 % 의 5-HMF 수율을 얻을 수 있었을 뿐만 아니라, 부반응물도 전혀 생성되지 않았기에 매우 선택적인 촉매로 확인되었다. FT-IR 분석에 의하면, 이는 NbCl5의 온화한 루이스 산성도에 기인한다. 또한, 촉매로서 4-히드록시-2,2,6,6,-테트라메틸-피페리딘-1-옥실 (4-히드록시-TEMPO) 라디칼을 보조 산화제로서 [비스(아세톡시)-아이오도]벤젠 (BAIB), 아세트산 (CH3COOH)을 사용하여 5-HMF의 2,5-DFF로의 호기성 산화반응을 연구하였다. 반응온도 30 oC 및 최적화된 TEMPO/BAIB/CH3COOH 조건하에서 45분만에 에틸 아세테이트 상에서 65.0 % 의 2,5-DFF 수율을 얻을 수 있었다. BAIB로 인하여 TEMPO의 히드록실아민 형태를 반응 중에 재생할 수 있었으며, 아세트산 첨가는 5-HMF 산화의 활성상인 옥소암모늄 형태의 TEMPO로의 변환을 촉진하였다. 본 반응시스템은 금속을 사용하지 않았기에 친환경적이며 에너지 효율적이다. TEMPO 촉매의 반복 회수 및 재생을 위하여 4-히드록시-TEMPO를 SBA-15에 고정화하였으며 (TEMPO-SBA-15), 제조된 촉매의 특성은 XRD, 질소흡탈착 등온곡선, TGA, SEM 및 FT-IR 등을 통해서 분석하였다. 5-HMF로부터 2,5-DFF 전환용 산화반응 촉매로 TEMPO-SBA-15, 보조 산화제로서 BAIB 및 아세트산을 사용한 결과 40 oC의 에틸 아세테이트 상에서 72.5 % 의 2,5-DFF수율을 얻었다. SBA-15의 기공 크기 분포 및 구조적 특성이 반응물과 생성물의 확산 이동을 촉진시켜 선택성이 향상된 것으로 판단된다. 뿐만 아니라 촉매는 여러 번의 반복 회수 및 재생 시에도 활성이 손실되지 않았다. 자성 나노입자가 실리카로 밀폐된 신규 이원기능성 촉매 (WO3HO-VO(Salten)-SiO2@Fe3O4)를 제조하기 위해서 Fe3O4 나노입자 표면을 실리카로 코팅한 후, 그 표면에 옥소바나듐 복합체 및 텅스텐산을 고정하였다. 제조된 촉매와 전구체들은 FT-IR, XRD, HR-TEM 질소 흡탈착 등온곡선 및 열중량분석 (TGA) 등을 통해서 분석되었다. WO3HO-VO(Salten)SiO2@Fe3O4 촉매는 과당으로부터의 탈수 및 산화반응의 두 단계에 모두 작용함으로써 과당으로부터 단일공정으로 2,5-DFF를 생성할 수 있게 하였다. 이를 통해서 중간화합물인 5-HMF의 복잡한 분리 및 정제 작업을 피할 수 있었다. 50 oC의 온화한 반응조건에서 과당으로부터 70.4 % 수율의 2,5-DFF를 단일공정으로 얻을 수 있었으며, 반응 후에는 외부자성체를 이용하여 촉매를 쉽게 회수할 수 있었다.

D-fructose가 흰쥐 소장으로부터 분리된 Brush-border membrane vesicles내로의 철의 취입에 미치는 영향

김옥선 全南大學校 1995 국내석사

천연당류중에서 철흡수를 촉진시키는 흡수촉진제를 개발하고자 위장관내에서 위장관 세포내로의 철의 취입(uptake)에 미치는 천연당류의 영향을 다소 수정된 2가 양이온 침전법에 따라 흰쥐의 소장으로부터 제조된 Brush-Border Membrane Vesicles(이하 BBMVs)를 사용하여 시험관내 취입 실험을 실시하였다. 그리고 천연당류중에서는 우선 먼저 단당류인 fructose를 선택하여 철과의 복합체 용액을 형성시키고 그들의 BBMVS내로의 취입 양상을 이미 철흡수를 증가시킨다고 잘 알려진 아스코르빈산과 철과의 복합체와 비교·검토함으로써 그들의 철흡수 촉진효율성을 검증하고자 하였다. 다소 수정된 2가 양이온 침전법에 따라 실험실적으로 제조된 BBMVs의 BBM의 기능 유지 확인방법을 표지 효소인 alkaline phosphatase의 활성도 측정과 전자 현미경법 및 D-glucose 취입을 통한 기능보존 측정법 등을 사용하여 검증 하였고, 철과 아스코르빈산 및 fructose등과의 복합체 용액을 제조하여(1:1000) 급속여과방법에 따라 이용하여 흰쥐 소장으로부터 제조된 BBMVs내로 취입 실험 및 농도 의존성 실험을 행하였다. 그리고 철과 fructose의 몰비율에 따라 취입속도가 어떻게 달라지는지를 알아 보았으며, 가장 높은 취입을 나타내는 비율에서 취입실험을 다시 행하였다. Alkaline phosphatase의 활성도 측정결과 16배 정도의 정제도를 나타냈고, D-glucose 취입실험에서는 전형적인 overshooting 취입현상이 나타났다. 전자현미경상의 BBMVs는 크기가 매우 일정하였고, 오염물질을 거의 찾아볼수 없었던 바, 수정된 2가 양이온 침전방법은 매우 정제도가 높은 방법으로써 철취입실험에 적합함을 알 수 있었다. 철복합물의 실험에서 Fe(III)-fructose가 Fe(III)-ascorbate보다 더 높은 Vmax값과 더 낮은 Km값 및 높은 취입속도를 보였다. 그리고, 철과 fructose를 일정 몰비율로 제조하여 취입 속도를 구한 결과 1:3의 비율에서 가장 취입속도가 높았다. 이상의 결과로 부터 fructose가 ascorbate보다 철의 흡수를 더 크게 촉진시킬 것으로 생각 되었고 또한 철과 fructose 사이에는 최적의 배합비율(1:3)이 존재함을 알 수 있었다. We have studied the iron uptake by the purified brush-border membrane vesicles (BBMVs) to determine the effect of fructose on the absorption of iron. BBMVs were prepared by the modified calcium precipitation method. The degree of purification was routinely assessed by the marker enzyme, alkaline phosphatase, the functional integrity was tested by D-[1-^3H]glucose uptake, and the appearance of membrane vesicles was shown by transmission electron microscopy (TEM). The uptakes of complexes of labeled iron [^(59)Fe] with fructose and ascorbic acid were measured with a rapid filtration technique. And the uptake rate and pattern of the two iron-complexes, Fe (III)-fructose and Fe(III)-ascorbate, were observed. In the study of dependency of iron uptake on concentration, two iron-complexes were diluted to 7.5-210 μM and the results were shown by the Lineweaver-burk plot. The Uptake rate of Fe(III)-fructose in various molar ratio of iron to fructose, was also measured, In results, A typical overshooting uptake of D-glucose was observed with peak values 2∼3 times the equilibrium. This result was similar to other studies with BBMVs; TEM showed that the size of BBMVs was uniform and we can hardly find any contaminants. In the uptake of iron complexes, the uptake amount and initial uptake rate were higher in Fe (III)-fructose than in Fe(III)-ascorbate. And Fe(III)-fructose has the higher value of Vmax and the lower value of Km than those of Fe (III)-ascorbate. In the study of molar ratio, the uptake rate of Fe (III)-fructose was highest in molar ratio of 1:3. It can be concluded that D-fructose is more effective in promoting the iron absorption than ascorbic acid and the optimum molar ratio of iron to fructose(1:3) exist.

Fructose-responsive regulation by FruR in Faecalibacterium prausnitzii for its intestinal colonization

최혜림 서울대학교 대학원 2025 국내박사

Faecalibacterium prausnitzii, a dominant member of healthy human gut microbiota, exhibits a strong positive correlation with fecal fructose levels, suggesting fructose as a key energy source for its colonization and persistence. This study explores the regulatory mechanisms governing the fru operon in F. prausnitzii, responsible for fructose uptake and metabolism. Here, we demonstrate that FruR, a DeoR family transcriptional regulator, orchestrates fru operon expression through interactions with fructose-1-phosphate (F1P) and HPr2, the histidine containing phosphocarrier protein. The F1P-HPr2(Ser-P)-FruR complex enhances RNA polymerase binding to the fru promoter, with stronger affinity for specific operator motifs compared to apo-FruR. F1P induces structural modifications in FruR that strengthen its interaction with HPr2 and alter its DNA recognition pattern, facilitating RNA polymerase access to the promoter. In vivo experiments in mice demonstrate increased F. prausnitzii abundance alongside upregul ated fru operon expression in fructose rich environments. This study provides new insights into how fructose availability modulates fru operon regulation and promotes F. prausnitzii colonization in the host intestine. Faecalibacterium prausnitzii는 건강한 인체 장내에서 우점하는 유익균으로 알려져 있지만, 산소에 매우 민감하고 배양이 까다로운 이 종이 다른 균들에 비해 장내정착이 유리한 원인에 대해서는 아직 연구되지 않았다. F. prausnitzii의 장내정착 수준은 분변 내 과당수준과 강한 양의 상관관계를 가지고 있음이 알려져 있으며, 이는 F. prausnitzii가 장내정착과 생장에 과당을 활발하게 이용할 것임을 시사한다. 본 연구에서는 F. prausnitzii에서 과당을 수송하고 대사하는 데에 필요한 유전자를 암호화하고 있는 fru 오페론의 전사조절 기작을 확인하였다. DeoR 구조를 가지는 전사인자인 FruR이 fructose-1-phosphate (F1P)와 histidine을 포함하는 인산기 전달 단백질인 HPr2와 결합하여 fru 오페론을 조절함을 밝혔다. F1P-HPr2(Ser-P)-FruR 복합체는 RNA polymerase가 fru promoter에 결합하는 것을 촉진시키며 apo-FruR과는 다른 양상으로 결합함을 볼 수 있었다. fru promoter에는 FruR이 결합할 수 있는 다수의 motif 가 있는데 F1P는 FruR이 해당 motif들에 결합하는 패턴을 변화시키며 더불어 FruR과 HPr2의 결합을 강화할 수 있음을 확인하였다. HPr2는 RNA polymerase가 fru promoter에 정상적으로 결합하는 도와주는 전사인자의 coactivator로 작용하며 FruR 전사활성 기작에 필수적인 인자임을 밝혔다. 나아가, 마우스를 이용한 in vivo 실험에서도 과당이 풍부한 장내환경에서 F. prausnitzii의 정착수준이 fru 오페론의 전사활성과 더불어 증가함을 확인하였다. 본 연구는 fru 오페론의 과당이 존재하는 환경을 인식하여 전사활성을 이루는 기작과 이에 따른 F. prausnitzii의 인체 장내정착에 대한 이해를 도울 수 있다.

Potential of microRNA-33 as a serological biomarker for fructose-induced liver injury

HanvitCha 고려대학교 대학원 2022 국내박사

과당으로부터 유발되는 비알코올성 지방간질환(Non-alcoholic fatty liver disease, NAFLD)의 병리학적 원인과 miRNA와의 상호 관련성에 대한 연구가 많이 부족하기 때문에 우리는 단백체학 분석을 통한 과당유래 비알코올성 지방간질환에서의 miRNA의 관련성 및 타겟을 발굴하고자 한다. 실험 I에서는 C57BL/6N 마우스에 물 또는 34% 과당을 자유식이를 통해 6주간 투여하여 비알코올성 지방간질환 모델을 유도하였고, 실험 II에서는 동일한 조건을 이용하여 4주 6주에서 희생하였으며, 시간에 따른 과당의 악영향을 관찰하였다. 실험 1의 결과에서 간 단백질체학에 대한 생물정보학 분석을 통해 SREBP1이 과당에 의해 영향을 받는 가장 중요한 상위 전사조절인자인 것으로 밝혔고, 이를 조절하는 miR-33-5p(miR-33)가 과당 섭취 시 SREBP1 조절을 담당하는 주요 miRNA로 확인되었다. 실험 II에서 우리는, 마우스가 과당을 오래 섭취할수록 더 심각한 간 손상 마커(예: 혈청 AST)가 더욱 증가하는 것을 확인 하였으며, 또한 Srebp1 mRNA 발현도 과당 섭취 기간에 따라 증가하였다. 반면에, 간 miR-33은 과당에 의해 시간 의존적으로 감소하였으나, 오히려 혈청 miR-33 발현은 증가하였다. 이러한 결과는 간에서 miR-33이 세포 손상 시 방출될 수 있음을 시사하였고, 마지막으로 우리는 fructose-induced ferroptosis가 산화 손상으로 인한 간 독성의 원인이 될 수 있음을 관찰하였다. 종합적으로, 우리의 연구 결과는 과당으로 인한 간의 산화 손상이 ferroptosis를 유도하고, 이에 따른 miR-33이 혈액으로의 유출이 일어나, 결과적으로 NAFLD의 혈청학적 바이오마커로서의 가능성을 밝혔다.

당질의 종류가 정상인과 당뇨병환자의 혈청 Glucose 및 혈청 Fructose에 미치는 영향

김혜숙 淑明女子大學校 1984 국내석사

Fructose has been claimed to offer advantage for diabetic individuals since it produces less hyperglycemia. However, previous studies have focused only on serum glucose responses, and thus do not address to the effect on serum fructose. This study was performed to investigate the effect of various simple sugars on serum glucose and fructose in normal and diabetic subjects. Subjects were given either 75g glucose, 75g fructose, 75g sucrose or 112.5g fructose in fasting state and their serum glucose and fructose levels were measured at 0,1,2 and 3 hours. The results were as following : 1) Fructose ingestion resulted in significantly lower serum glucose than ingestion of either sucrose or glucose both in normal and diabetic subjects. 2) Increment in serum fructose was highest after ingestion of fructose compared to ingestion of sucrose or glucose both in normal and diabetic subjects. 3) In normal subjects, serum fructose level increased as the dose of fructose in creased. 4) In diabetic subjects, serum fructose level remained at high level and became significantly higher than normal subjects after 3 hours. 5) Fructose, sucrose and glucose were compared for its possible biohazard (△ glucose plus △ fructose x 7.5) and sucrose gave highest numbers in diabetic subjects. There result raise possibility that ingestion of fructose and sucrose can produce undesirable effects in diabetics.

Allose biosynthesis of fructose by one-pot reaction with Flavonifractor plautii D-allulose 3-epimerase and Clostridium thermocellum ribose 5-phosphate isomerase

이태의 건국대학교 대학원 2018 국내석사

D-Allose is a potential medical sugar because it has benefit pharmaceutical effects, including anti-cancer, anti-hypertensive, anti-inflammatory, anti-oxidative, anti-tumor, cryoprotective, and immunosuppressant activities. One-pot reaction using Flavonifractor plautii D-allulose 3-epimerase (FP-DAE) and Clostridium thermocellum ribose 5-phosphate isomerase (CT-RPI) produced allose from fructose, an inexpensive substrate. For the production of allose from fructose, CT-RPI was used along with FP-DAE. The optimal reaction conditions were pH 7.5, 60 °C, 0.1 g/l FP-DAE, 12 g/l CT-RPI, and 600 g/l fructose in the presence of 1 mM Co2+. Under these optimized conditions, FP-DAE and CT-RPI produced 79 g/l allose for 2 h, with a conversion yield of 13%. This is the first biotransformation of fructose to allose by a two-enzyme system. The production of allose by one-step reaction using FP-DAE and CT-RPI was increased by 130%, comparing with that by two-step reaction using the two enzymes. D-allose는 항암, 항고혈압제, 항염증제, 항산화제, 항종양제, 동결방지제, 면역억제제 등의 제약 효능이 있기 때문에 잠재적인 medical sugar라고 할 수 있다. 현재까지 allose는 allulose로부터 생산되어 왔다. 하지만 allulose는 allose보다 비싸다는 큰 문제가 있다. 때문에 allulose로부터 allose를 생산하는 것은 산업적으로 장점이 없다. 하지만 Flavonifractor plautii 유래의 D-allulose 3-epimerase (FP-DAE)와 Clostridium thermocellum 유래의 ribose 5-phosphate isomerase (CT-RPI)를 혼합하여 one-pot 반응을 하면, 값싼 기질인 fructose로부터 allose를 생산할 수 있다. 0.1 g/l FP-DAE와 12 g/l CT-RPI를 1 mM의 Co2+와 pH 7.5, 60 °C에서 600 g/l의 fructose를 기질로 사용했을 때가 최적조건이다. 위에서 언급한 최적 조건에서 FP-DAE 및 CT-RPI는 2시간 동안 79 g/l의 allose를 생산하였으며, 전환율은 13 %였다. 이것은 two-enzyme system에 의해 fructose로부터 allose로의 최초 생물학적 전환이다. FP-DAE와 CT-RPI를 이용하여 fructose로부터 allose를 생산하는데 있어서 one-step 또는 two-step 반응이 있으며 이 두가지 방법을 비교하였을 때, one-step reaction이 130 %로 더 많은 전환이 된 것을 확인하였다.

High Fructose Intake Promotes Adipose-Lung Crosstalk via Inflammatory Mediators, Exacerbating Allergic Asthma Responses in OVA- Sensitized C57BL/6 Mice

Gamaralalage Prabhavi Dayaddra Dassanayake 제주대학교 대학원 2025 국내석사

Dietary fructose is increasingly recognized as a potent immuno-metabolic stressor. However , its mechanistic contribution to the pathogenesis of airway allergy remains inadequately characterized. This study elucidates how chronic fructose consumption amplifies allergen-driven inflammation in an ovalbumin (OVA)-sensitized C57BL/6 murine model. A high-fructose diet substantially elevated lipid peroxidation in pulmonary and adipose tissues, reflecting a pronounced reactive oxygen species (ROS) generation and systemic oxidative burden. This metabolic dysregulation coincided with marked epithelial architectural remodeling, mucus hypersecretion, and extensive infiltration with mononuclear and polymorphonuclear cells. Fructose-exposed mice demonstrated GLUT5 and GLUT2 transcriptional upregulation, indicating augmented fructose transmembrane transport and intracellular metabolic flux. Immunohistochemical analyses revealed substantial F4/80⁺ and CD11c⁺ macrophage enrichment with pronounced M1-dominant CD80⁺ expression and diminished CD206⁺ M2 macrophage population. These M1 macrophages exhibited heightened phagocytic activity, supported by transcriptomic enrichment of phagosome maturation, lysosomal fusion, and myeloperoxidase (MPO)-dependent oxidative pathways. MPO was markedly elevated, and DIF revealed widespread extracellular chromatin scaffolds. MPO co-localization with citrullinated histone H3 (CitH3) confirmed both neutrophil extracellular traps (NETs) and macrophage extracellular traps (METs), with MET formation disproportionately prominent in fructose groups. Chemokine profiling demonstrated robust CCL2, CCL5, CXCL1, CXCL2, CXCL5, and CXCL9 upregulation across both tissues, substantiating the bidirectional adipose-lung inflammatory crosstalk. Transcriptomic analyses confirmed enrichment of ROS biosynthesis, neutrophil degranulation, phagosomal maturation, and chemokine-driven leukocyte trafficking pathways. Thus, dietary fructose exacerbates allergic airway inflammation by intensifying oxidative stress and promoting M1-dominant macrophage activation with amplified phagocytic activity, prominent NETosis and METosis, and fortified adipose-lung chemokine communication. Therefore, fructose serves as a critical dietary determinant with significant implications for asthma pathogenesis and therapeutic intervention.

Light dependent expression of pea chloroplast fructose-1,6-bisphosphatase

Im, Chae-hwan 慶熙大學校 1992 국내석사

Purification and characterization of chloroplast fructose-1, 6-bisphosphatase from pea leaves : 완두 엽록체 Fructose bisphosphatase의 분리정제 및 특성

부성희 Graduate School Kyung Hee University 1990 국내박사

Pentoxifylline과 단당류 fructose와 galactose가 정자운동에 미치는 영향

편정환 경북대학교 대학원 2018 국내석사

Sperm motility is an important factor in fertilization and pregnancy. The sperm viability of commercial cow frozen semen and boar liquid semen tends to decrease after freezing compared with before freezing and as the sperm storage time increases, respectively. Before in vitro fertilization or intrauterine insemination, pentoxifylline as a cell signaling pathway factor can be used to improve sperm motility. Pentoxifylline inhibits phosphodiesterase activity and continuously increases intracellular cAMP levels. This increased cAMP promotes the activity of enzymes involved in the production of ATP required for sperm motility. Therefore, pentoxifylline is thought to be able to improve sperm motility through cell signaling pathways. In addition, monosaccharides like fructose and galactose as the energy sources to produce ATP is thought to be able to improve sperm motility. In other words, pentoxifylline(PTX) and monosaccharide when added to media for sperm incubation is thought to be able to improve sperm motility. In order to propose a culture condition to improve the motility of bull and boar sperm, this study investigated the effect of pentoxifylline and monosaccharides fructose and galactose on sperm motility. Cow frozen semen and boar liquid semen was used to analyze the effect of PTX and monosaccharide on sperm motility with sperm motility index and computer assisted sperm analyser(CASA) method. The effect of PTX treatment on cow frozen semen was investigated, 5mM PTX significantly increased sperm motility(MOT) compared to control. The effect of monosaccharide on sperm motility was investigated, 10mM fructose increased sperm motility VAP, ALH and BCF, but galactose did not affect sperm motility. In addition, the effect of PTX and monosaccharide combined treatment on bull sperm motility was investigated, they did not increased sperm motility. The effect of PTX treatment on boar liquid semen was investigated, 5mM PTX significantly increased the sperm motility compared to other treatments, especially at the culture for 40 minutes. The effect of monosaccharide on sperm motility was investigated, 10mM fructose significantly increased sperm motility VAP and 20mM fructose significantly increased sperm motility VCL, compared to the 5mM frcutose, respectively. However, 20mM galactose significantly increased sperm motility VSL compare to control, especially at the cultures for 40minutes and 50minutes. In addition, the effect of PTX and monosaccharide combined treatment on boar sperm motility was investigated, Of all treatments, sperm motility VCL and VSL were increased only in 20mM PTX and 10mM galactose combined treatment. In conclusion, PTX inproved sperm movement in cow frozen semen and boar liquid semen. Especially, PTX 5mM may be a optimal concentration for the sperm motility. On the other hand, 10mM fructose or 20mM galactose may increase effectively the sperm motility in boar liquid semen. The combined treatment of 20mM PTX and 10mM galactose can improve the sperm motility in the boar semen. Therefore, in order to improve the movement of sperm in frozen and liquid semen, it is recommended to add 5mM PTX to the media or diluent at 40 minutes before use.