• 한국축산식품학회지
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• 제37권3호
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• pp.402-409
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• 2017

A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

• Journal of Acupuncture Research
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• 제33권4호
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• pp.49-63
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• 2016

Objectives : This research was performed to investigate the effects of Jodeungsan pharmacopuncture(PA-J) of focal brain ischemia induced by middle cerebral artery occlusion(MCAO) in rats. Methods : The subjects were divided into 4 groups : control, acupuncture, pharmacopuncture PA-J1(11.43 mg / 250 g / $40{\mu}{\ell}$) and pharmacopuncture PA-J2(2.29 mg / 250 g / $40{\mu}{\ell}$). The focal brain ischemia was induced by intraluminal filament insertion into the middle cerebral artery. After 3 days of MCAO, Jodeungsan pharmacopuncture treatment was performed on the GB20, and the day after being treated with pharmacopuncture, the Morris water maze test was carried out on the assigned group. The series of processes were administered 6 times. Thereafter mGluR5, density of neuronal cell and ChAT were measured. Results : The results were as follows. 1. The distance to target significantly decreased in the 2nd trial of the Acu group on the water maze test for short-term memory. 2. The distance to target significantly decreased in the 4th trial of the PA-J2 group on the water maze test for long-term memory. 3. The intensity of mGluR5 significantly increased in the PA-J1 group compared with the control group. 4. The neuroprotective effect on the hippocampal CA1 significantly increased in the PA-J1 and PA-J2 groups compared with the control group. 5. The density of ChAT in the hippocampal CA1 significantly increased in the PA-J1 and PA-J2 groups compared with the control group. Conclusion : These results suggest that Jodeungsan pharmacopuncture may improve memory and cognitive impairment and also have neuroprotective effects on focal brain ischemia.

• 한국토양비료학회지
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• 제45권5호
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• pp.787-791
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• 2012

펩타이드는 Asp, Glu, Leu을 주요 구성 아미노산으로 하는 한우 혈청을 단백질 분해효소로 분해 후 한외여과를 거쳐 제조하였다. 단백질 분해효소는 alcalase, esperase, flavourzyme, protamex를 이용하였고 alcalase가 가장 좋은 분해력을 나타내어 폡타이드 생산을 위한 혈청 분해에 활용하였다. 한우 혈청으로부터 분리한 펩타이드와 킬레이트 결합한 칼슘 (chelate-Ca)과 게르마늄 (chelate-Ge)을 방울토마토에 적용하여 흡수 변화를 확인하고자 시험을 수행하였다. Chelate Ca을 500, 1,000, 2,000배로 희석하여 방울토마토 재배에 적용한 결과, 처리 농도 및 횟수 증가에 따라 엽과 과실의 칼슘함량이 다소 증가하여 대조처리 Calciolid Ca-300 (3409.1, $67.5mg\;kg^{-1}$)이 Chelate Ca-200 (3781.1, $78.0mg\;kg^{-1}$) 보다 처리농도가 높음에도 불구하고 그 함량이 낮아 킬레이트 칼슘의 흡수율이 더 많았다. Chelate Ge을 50, 100, 200배로 희석하여 적용한 결과, 방울토마토 엽의 게르마늄 함량이 처리 농도가 높을수록 많아져 대조구와 Ge $132^{(R)}$-20 보다 모든 처리구에서 2배 이상의 높은 함량을 나타내었다. 본 연구를 통해 펩타이드와 킬레이트 결합한 칼슘과 게르마늄을 방울토마토에 시용할 경우 그 흡수율이 증가되는 것을 확인하였다.

• Journal of Acupuncture Research
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• 제31권4호
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• pp.57-70
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• 2014

Objectives : The purpose of this study is to examine if Clematidis Radix(CR) pharmacopuncture may be effective to the neuropathic pain(mechanical allodynia) in a rat model of neuropathic pain. Methods : To produce the model of neuropathic, L5 spinal nerve was ligated by 6-0 silk thread. After neuropathic surgery, the author examined if the animals exhibited the plantar withdrawal response of allodynia. The plantar withdrawal response was assessed by dynamic plantar aesthesiometer three days after the neuropathic surgery, CR pharmacopuncture was injected at $BL_{23}$ 1time/week for 6 weeks. After that, the author examined the plantar withdrawal response of rats' leg by dynamic plantar aesthesiometer. And also the author examined mGluR5, Bax, Bcl-2, Bax/Bcl-2 ratio in spinal cord, and c-Fos. Also the author observed the change of aspartate aminotransferase(AST), alanine aminotransferase(ALT) count in the blood serum of neuropathic rats. Results : 1. The withdrawal response of allodynia decreased in the PT3, PT4 group as compared with control group. 2. The mGluR5 increased in the PT1, PT2, PT3, and PT4 group. 3. The Bax and Bax/Bcl-2 ratio decreased in the PT4 group. 4. The c-Fos increased in the PT1 group, and decreased in the PT4 group. 5. The changes of AST in blood serum decreased in the every group excluded control group and the changes of ALT in blood serum isn't shown the signigicant change. Conclusions : These results are suggested that CR pharmacopuncture at BL23 decreased mechanical allodynia, can act on anti-apoptotic, neuroprotective effects and liver fuction in the model of neuropathic pain.

• Fisheries and Aquatic Sciences
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• 제14권3호
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• pp.174-178
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• 2011

This study aimed to determine the degree of hydrolysis and angiotensin-I-converting enzyme (ACE)-inhibitory activity of Giant Jellyfish Nemopilema nomurai (jellyfish) hydrolysates. The degree of hydrolysis using six proteolytic enzymes (Alcalase, Flavozyme, Neutrase, papain, Protamex, and trypsin) ranged from 13.1-36.8% and the inhibitory activities from 20.46-79.58%. Using papain hydrolysate, we newly isolated and characterized ACE-inhibitory peptides with a molecular weight of 3,000-5,000 Da that originated from jellyfish collagen. The purified peptide (FII-b) was predicted to be produced from an alpha-2 fragment of the type IV collagen of jellyfish. The N-terminal sequence of FII-b was Asp-Pro-Gly-Leu-Glu-Gly-Ala-His-Gly- and showed 87% identity to the collagen type IV alpha-2 fragment of Rattus norvegicus and a predicted protein from Nematostella vectensis, indicating that the ACE-inhibitory peptide originated from the collagen hydrolysate and had an $IC_{50}$ value of 3.8 ${\mu}g$/mL. The primary structure of the fragment is now being studied; this peptide represents an interesting new type of ACE inhibitor and will provide knowledge of the potential applications of jellyfish components as therapies for hypertension.

• Food Science and Biotechnology
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• 제17권3호
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• pp.489-494
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• 2008

In order to develop a potent antidementia $\beta$-secretase inhibitor from phytochemicals, $\beta$-secretase inhibitory activities of extracts from many medicinal plants and herbs were determined. Water extracts from Rubus coreanus showed the highest $\beta$-secretase inhibitory activity of 84.5%. After purification of the $\beta$-secretase inhibitor from R. coreanus using systematic solvent extraction, ultrafiltration, Sephadex G-10 column chromatography, and reverse-phase high performance liquid chromatography (HPLC), a purified $\beta$-secretase inhibitor with $IC_{50}$ inhibitory activity of $6.3{\times}10^3\;ng/mL$ ($1.56{\times}10^{-6}\;M)$ was obtained with a 0.08% solid yield. The molecular mass of the purified $\beta$-secretase inhibitor was estimated to be 576 Da by liquid chromatography-mass spectrometry (LC-MS) and $\beta$-secretase inhibitor also is a new tetrapeptide with the sequence Gly-Trp-Trp-Glu. The purified $\beta$-secretase inhibitory peptide inhibited $\beta$-secretase non-competitively and also show less inhibition on trypsin, however no inhibition on other proteases such as $\alpha$-secretase, chymotrypsin, and elastase.

• Journal of Acupuncture Research
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• 제25권1호
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• pp.1-14
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• 2008

Objectives : Delayed neuronal death(DND) of pyramidal neuronsin the CA1 regions of the hippocampus has been extensively studied following global brain ischemia, whereas little is known about DND in this highly vulnerable brain region after focal brain ischemia. The aim of the present study was to investigate the effect of Gastrodiae Elata(GA) pharmacopuncture on hippocampal neuronal apoptosis in rats with focal brain ischemic injury. Materials and methods : The neuroprotective effects of water extracts of GA were investigated in middle cerebral artery occlusion(MCAo) of Sprague-Dawley(SD) rats. Seventy-five healthy SD ratswere randomly divided into five groups following MCAo : control group with focal ischemia, saline injection group, pharmacopuncture group GA-1($0.0007mg/m{\ell}/g$), pharmacopuncture group GA-2($0.00035mg/m{\ell}/g$), pharmacopuncture group GA-3($0.00014mg/m{\ell}/g$). Results : The intensity of mGluR5 increased in the GA-1 group. The intensity of Bax and the Bax/Bcl-2 ratio decreased in the GA-1 group. The intensity of Bcl-2 increased in all the GA groups. The density of neurons stained by Cresyl violet and ChAT increased in the GA-1 group. Conclusions : Our study suggests that GA pharmacopuncture at $GB_{20}$ showed anti-apoptotic and neuroprotective effects on cholinergic neuronsin focal cerebral ischemia caused by stroke in SD rats.

• Korean Journal of Acupuncture
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• 제23권3호
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• pp.81-98
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• 2006

Objectives : Acupuncture using a tonification or sedation techniques method is used as a controlling the medication for an early stroke in the Korean medicine. LR3 has indicatons of headache, vertigo, facial paralysis, apoplexy, epiepsy as the source acupoint of a liver meridian. So this study is aims to investigate the anti-apoptotic and neuroprotective effects of acupuncture on the focal ischemia induced by intraluminal filament insertion in rats. Methods : The focal ischemia was induced by intraluminal filament insertion into middle cerebral artery. The animals were divided into seven groups (n=8 in each group) : Normal, intactness group; Conrol group, no therapy group after being ischemia induced; MA-l, acupuncture perpendicularly without Tonification or Sedation techniques at LR3 after being ischemia induced; MA-2, acupuncture obliquely towards the knee at LR3 after being ischemia induced; MA-3, acupuncture obliquely towards the toe at LR3 after being ischemia induced; MA-4, acupuncture obliquely towards the knee and rotate 9 times in a clockwise direction at LR3 after being ischemia induced; MA-5, acupuncture obliquely towards the toe and rotate 6 times in a counterclockwise direction at LR3 after being ischemia induced. The anti-apoptotic and neuroprotective effects of Acupuncture techniques of tonification or sedation at LR3 are observed by mGluR5, Bax, Cresyl violet, ChAT-stain and NGF. Results : The intensity of mGluR5 and the density of ChAT was increased in MA-1 group. The intensity of Bax was decreased in MA-3, MA-4 group. The density of neurons stained by Cresyl violet and ChAT was increased in MA-2, MA-3, MA-4, MA-5 group. The density of neurons stained by NGF was only increased in MA-4 group. Conclusions : Our study suggests that acupuncture perpendicularly without Tonification or Sedation techniques and obliquely towards the knee and rotate 9 times in a clockwise direction(Tonifying technique) at LR3 after being ischemia induced at LR3 shows anti-apoptotic and neuroprotective effects on cholinergic neuron in focal cerebral ischemia of the stroke in rats.

• Animal cells and systems
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• 제11권2호
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• pp.175-182
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• 2007

The lysozyme II gene of cabbage butterfly Artogeia rapae was cloned from fat body of the larvae injected with E. coli and its nucleotide sequence was determined by the RACE-PCR. It has an open reading frame of 414 bp nucleotides corresponding to 138 amino acids including a signal sequence of 18 amino acids. The estimated molecular weight and the isoelectric point of the lysozyme II without the signal peptide were 13,649.38 Da and 9.11, respectively. The A. rapae lysozyme II (ARL II) showed the highest identity (81%) in the amino acid sequence to Manduca sexta lysozyme among other lepidopteran species. The two catalytic residues ($Glu^{32}$ and $Asp^{50}$) and the eight Cys residue motifs, which are highly conserved among other c-type lysozymes in invertebrates and vertebrates, are also completely conserved. A phylogenetic analysis based on amino acid sequences indicated that the ARL II was more closely related to M. sexta, Hyphantria cunea, Heliothis virescens, and Trichoplusia ni lysozymes. The ARL II gene was expressed in Spodoptera frugiperda 21 insect cells and the recombinant ARL II (rARL II) was purified from cell-conditioned media by cation exchange column chromatography and reverse phase FPLC. The purified rARL II was able to form a clear zone in lysoplate assay against Micrococcus luteus. The lytic activity was estimated to be 511.41 U/mg, 1.53 times higher than that of the chicken lysozyme. The optimum temperature for the lytic activity of the rARL II was $50^{\circ}C$, the temperature dependency of the absolute lytic activity of rARL II was higher than that of the chicken lysozyme at low temperatures under $65^{\circ}C$.

• 한국수산과학회지
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• 제26권5호
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• pp.416-423
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• 1993

수산자원의 고도이용과 기능특성 해명을 위한 연구의 일환으로 수산식품 중에 존재하는 생리활성물질을 검색하기 위하여 전통수산발효식품의 하나인 정어리 어간장의 숙성에 따른 ACE 저해작용 및 gel 여과에 의한 활성획분의 분리 및 그 특성을 살펴본 결과는 다음과 같다. 1. 정어리 어간장의 숙성에 따른 $5\%$ TCA 가용성 peptide-nitrogen의 생성은 전반적으로 숙성기간의 경과와 더불어 증가하다가 감소하는 경향을 나타내었으나 잔사를 이용한 정어리 어간장의 경우는 숙성과 더불어 계속 감소하는 경향을 나타내었다. ACE 저해작용 또한 peptide-nitrogen의 생성과 유사한 경향을 나타내었으며, 코오지 첨가구가 무첨가 경우보다 다소 높은 것으로 나타났으나 잔사를 이용한 경우는 오히려 코오지 무첨가 경우가 약간 높은 것으로 나타났다. 2. 정어리 어간장의 ACE 저해효과는 첨가량에 따라 급격히 증가하는 것으로 나타났으며 특히 육으로 만든 정어리 어간장의 ACE 저해효과는 코오지 첨가구의 경우가 무첨가구에 비하여 높은 것으로 나타났다. 3. ACE 저해작용에 대한 가열의 영향은 다소차이는 있으나 거의 변화가 없는 것으로 나타났다. 4. $IC_{50}$은 코오지 무첨가 잔사, 육 및 전어체로 만든 정어리 어간장의 경우는 각각 $125{\mu}g,\;200{\mu}g$ 이상 $125{\mu}g$으로 나타났으며, 코오지 첨가구의 경우는 각각 $140{\mu}g,\;100{\mu}g$ 및 $135{\mu}g$으로 나타났다. 5. Gel 여과에 의한 정어리 어간장의 ACE 저해작용을 가지는 획분중 획분 C가 가장 우수한 것으로 나타났으며, 이 때의 분자량은 1,400전후인 것으로 나타났다. 6. 획분 C의 아미노산 조성은 Glu, Ala, Lys 및 Leu의 함량이 높은 것으로 나타났다.