카드뮴 노출에 따른 북방전복, Haliotis discus hannai (Ino, 1953) 의 hemolymph 및 hepatopancreas의 효소활성의 변화

민은영,이정식,곽인실,김재원,강주찬 한국패류학회 2014 The Korean Journal of Malacology Vol.30 No.1

This study was conducted to investigate the effects of cadmium (Cd) exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone, Haliotis discus hannai. The abalone were exposed to 0, 5, 10, 20 and 40 μg/L Cd for 4 weeks. The phenoloxidase (PO) activity was decreased in hemolymph of abalone exposed to 40 Cd μg/L for 4 weeks compared to the control (P < 0.05). The hemolymph enzymes, alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were markedly elevated in 40 Cd μg/L after 4 weeks. The hemolymph calcium concentrations were significantly decreased in 20 and 40 Cd μg/L for 4 weeks. Hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were significantly increased by Cd. SOD was increased in both 20 and 40 Cd μg/L and CAT, in 40 Cd μg/L after 2 weeks (P < 0.05). These results suggested that the abalone SOD and CAT including PO may serve as a protective mechanism against oxidative stress by Cd. We conclude that a Cd concentration, 40 μg/L in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone hepatopancreas. From these results, these biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas. 본 연구는 카드뮴 노출에 따른 북방전복, Haliotis discus hannai의 hemolymph 및 hepatopancreas의 효소활성에 대한 변화를 관찰하였다. 북방전복은 카드뮴 0, 5, 10, 20 및 40 Cd μg/L의 농도에서 4 주간 노출시킨 결과, hemolymph의phenoloxidase (PO) 활성은 40 Cd μg/L 농도에서 4주 후에 유의하게 감소하였으나, alkaline phosphatase (ALP), aspartate aminotransferase (AST) 및 alanine aminotransferase (ALT) 활성은 40 Cd μg/L 농도에서 4 주 후에 유의한 증가를 나타냈다. 또한 hemolymph의 칼슘(Ca) 농도는 20 및 40 Cd μg/L의 농도에서 4주 후에 유의한감소를 나타냈다. 한편 hepatopancreas의 superoxide dismutase (SOD) 활성은 20 및 40 Cd μg/L 농도에서, catalase (CAT) 활성은 40 Cd μg/L 농도에서 2주 후에 유의한 증가를 나타냈다. 따라서 카드뮴에 노출된 북방전복hemolymph와 hepatopancreas의 효소활성은 적어도 카드뮴농도 40 Cd μg/L에서 변동이 있을 것으로 예상되며, 이들 효소활성은 연안의 중금속 오염 모니터링의 기초 자료로 활용할수 있을 것으로 생각된다.

곤충세포 배지 개발을 위한 체액산화지연 돌연변이 누에계통 선발

최지영 ( Ji Young Choi ),김종길 ( Jong Gill Kim ),최영철 ( Young Cheol Choi ),윤형주 ( Hyung Joo Yoon ),안미영 ( Mi Young Ahn ),김삼은 ( Sam Eun Kim ),황석조 ( Seok Jo Hwang ) 한국잠사학회 2007 한국잠사곤충학회지 Vol.49 No.2

누에 체액은 공기 중의 산소와 접하게 되면 도파퀴논(dopaquinone)이라는 세포 성장을 저해하는 물질을 생성하게 된다. 체액의 산화를 방지하기 위해서는 일반적으로 열처리에 의해서 효소를 불활성화 시키는 방법이 이용되고 있으나 대량 채혈 시에는 열처리 이전에 이미 산화가 진행되는 문제점이 있다. 이러한 문제점을 해결하기 위하여 누에 중 체액 산화가 더디게 진행되는 누에 계통을 선발하게 되었다. 누에체액을 채취하여 경과시간별로 산화 정도(변색)를 육안으로 확인하여 TBO, Y4, 백안Eb과잉지 3품종을 선발하고 spectrophotometer를 이용하여 산화반응 속도를 측정하였다. 백옥잠 체액과 각각의 산화지연 누에 체액의 산화속도를 400 nm에서 측정한 결과 백옥잠은 120분인데 비하여 체액산화 지연계통인 Y4는 330분, TBO는 360분 그리고 백안Eb과잉지는 450분에 산화가 완료되었다. 산화지연 누에계통의 체액의 곤충세포 증식에 미치는 영향을 조사한 결과 대규모 채혈한 체액산화지연 계통의 체액은 야성형 계통(백옥잠)에서 소규모 채취한 체액에는 미치지 못하였으나 대규모 채취한 체액보다는 우수하였다. Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as Y4, TBO wEb and showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at 20℃ in each 330 min (Y4), 360 min (TBO) and 450 min (wEb) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.

TBTO의 노출에 따른 참굴, Crassostrea gigas의 hemolymph내 무기성분 및 효소활성의 변화

조규석,민은영,지정훈,김재원,안철민,강주찬 한국어병학회 2001 한국어병학회지 Vol.14 No.2

본 연구는 20일 동안 TBTO 노출에 의한 참굴의 생존율 및 hemolymph 내의 무기성분과 효소활성의 변동을 조사하였다. TBTO 노출에 의한 참굴의 생존율은 실험 20일 동안 10㎍/L이하의 농도에서 90%이상을 나타냈다. 그러나 참굴의 생존율은 TBT농도가 증가함에 따라 감소하기 시작하여 실험 10일째 TBT농도 80㎍/L이상, 실험 20일째는 50㎍/L이상의 농도에서 유의한 감소가 관찰되었고, 100㎍/L에서는 실험 16일째 전 개체가 사망하였다. 참굴의 hemolymph의 calcium 농도는 10일째 대조구를 포함한 TBTO 5∼10㎍/L 농도에서 유사한 값을 나타냈으나 50㎍/L 이상의 TBTO 농도에서는 급격히 증가하여 대조구에 비해 유의한 증가가 관찰되었다. 또한 20일째 calcium 농도는 10일째 보다 전반적으로 증가하였으며, 대조구와 비교하여 20일㎍/L 이상의 농도에서 유의한 증가를 나타냈다. GPT활성은 실험 전 기간을 통해 모든 농도구에서 뚜렷한 변화를 보이지 않았다. 그러나 GOT는 실험 10일째 이상의 20㎍/L이상의 농도에서 유의한 증가를 나타냈다. The purpose of this study is to find out the effects of various bis(tri-n-butyltin)oxide (TBTO) on changes of inorganic matter and enzyme activity in the hemolymph of Pacific Oyster, Crassostrea gigas. Oyster were exposed to 5,10,20,50,80and 100㎍/L of TBTO for 20 days. Survival rate of the oyster wass significantly affected by ≥80㎍/L TBTO concentration at 10 days, while the diminution of survival was found at 20 days with a lower concentration of ≥50㎍/L TBTO. Calcium concentration in the hemolymph increased significantly after 20 days at the TBTO concentration 20 ㎍/L. However no change of magnesium and inorganic phosphate in the hemolymph was showed. A significant increment of GOT activities in the hemolymph was observed after 20 days at more than 20㎍/L TBTO concentraion, without typical changes of GPT activities. These results indicate that oysters can be affected by TBTO in terms of calcium concentration and GOT activity in the hemolymph when they were exposed to the TBTO concentration 20㎍/L.

발효 누에 숙잠 혈림프의 항산화 및 항티로시나제 활성

문지영 ( Ji Young Mun ),조유영 ( You Young Jo ),권해용 ( Hae Yong Kweon ),이광길 ( Kwang Gill Lee ),여주홍 ( Joo Hong Yeo ),이희삼 ( Heui Sam Lee ) 한국잠사학회 2014 한국잠사곤충학회지 Vol.52 No.2

In this study, functionality of products of silkworm hemolymph fermented by Bacillus species was studied such as cell viability, antioxidant effect, and inhibitory effect on tyrosinase activities. A matured silkworm hemolymph was degraded by fermentation with Bacillus subtilis 10854 and Bacillus amyloliquefaciens M27. Especially, proteins of matured silkworm hemolymph were degraded to 3,000 Da by fermentation with B. amyloliquefaciens M27. Cell viability for MTT assay was higher than PBS in hemolymph and fermented hemolymph. A DPPH free radical scavenging activity of fermented hemolymph was as higher as Vitamin C and dependent on sample concentrations. Thus, these results suggest that degraded hemolymph fermented by B. amyloliquefaciens M27 may have antioxidant properties as a material for cosmetics.

수온 상승에 따른 북방전복, Haliotis discus hannai과 둥근전복, H. discus discus 체액의 생리학적 변화

민은영,이정식,김재원,전미애,강주찬,Min, Eun-Young,Lee, Jung Sick,Kim, Jae Won,Jeon, Mi Ae,Kang, Ju-Chan 한국패류학회 2015 The Korean Journal of Malacology Vol.31 No.1

본 연구에서는 수온 증가에 따른 북방전복 (Haliotis discus hannai) 과 둥근전복 (H. discus discus) hemolymph의 생리 및 면역학적 변화를 관찰하기 위하여 위 두 종을 20, 22, 24, 26 및 $28^{\circ}C$ 수온에 각각 4 일간 노출시켰다. 노출 결과, hemolymph의 total-protein (TP), glucose, 칼슘 (Ca) 은 둥근전복이 북방전복보다 높은 값을 보였으나, 마그네슘 (Mg), alkaline phosphatase (ALP) 및 lysozyme은 두 종에서 유사한 값을 보였다. 수온 증가에 따른 hemolymph의 TP, glucose 및 마그네슘은 두 전복 모두에서 유의한 변화가 관찰되지 않았다. 그러나 칼슘과 면역관련 인자인 ALP 및 lysozyme은 둥근전복에서 유의한 차이가 없었으나, 북방전복에서는 $26^{\circ}C$ 또는 $28^{\circ}C$의 고수온에 노출되었을 때, 유의하게 높은 활성을 보였다. 한편 phenoloxidase (PO)는 북방전복에서 높은 값을 보였으며, 수온이 높을수록 증가하는 경향을 보였다. 결론적으로 북방전복은 둥근전복과 달리 여러 지표에서 유의한 변화가 관찰된 것으로 보아 북방전복이 둥근전복보다 고수온에 보다 민감한 것으로 사료된다. This study was conducted to investigate the effects of elevated water temperature (WT) on biochemical and immunological factors in the hemolymph of the abalones, Haliotis discus hannai and H. discus discus. The abalone were exposed to various WT; 20, 22, 24, 26 and $28^{\circ}C$ for 4 days. In the control and $20^{\circ}C$, total-protein (TP), glucose and calcium (Ca) in hemolymph of H. discus discus were higher than the values in H. discus hannai. The values of magnesium (Mg), alkaline phosphatase (ALP) and lysozyme in H. discus hannai were similar to the H. discus discus in the control. There were no significant alterations in TP, glucose and Mg levels of hemolymph in H. discus hannai and H. discus discus by WT increases. The values of Ca, ALP and lysozyme were increased in H. discus hannai exposed to the high temperature (26 and $28^{\circ}C$) compared to control, while the values in H. discus discus were not significant difference between the WT groups. The phenoloxidase (PO) activity was increased in hemolymph of H. discus hannai exposed to high temperature (${\geq}24^{\circ}C$) compared to the control (P < 0.05). These physiological and immunological parameters were significantly changed in H. discus hannai. However, these parameters in H. discus discus were barely altered at the high WT (P < 0.05). These results suggested that H. discus hannai is considered to be more sensitive than H. discus discus at the high WT.

집파리유충 hemolymph 중신형의 anti-fungal peptides의 분리정제

Jian-Wei Wu,Li-juan Gu,Xiao-Qing Su,성창근 한국생명과학회 2006 생명과학회지 Vol.16 No.3

To isolate and purify anti-fungal active substances from immunized housefly (Musca domestica), low dose of Candida albicans was injected into the larvae of the housefly to induce the appearance of potent anti-fungal active substances in the hemolymph. This purification work was performed by the routine isolation and purification processes of protein, namely, solid phase extraction (SPE), SDS-PAGE electrophoresis, HPLC purification. Three 4~16 kDa peptides which exhibited antifungal activity against Candida albican and other fungi were isolated from induced hemolymph. Consequently, further anti-fungal activity study showed that these three peptides were different either in molecular weight or in anti-fungal activity. All isolated substances were proved to be active and resistant to high-temperature. It was deduced that these peptides isolated from induced housefly were novel members of the insect defensin family and they were inducible. 유도된 집파리유통 hemolymph중에서 Candida albicans의 3가지 anti-fungal peptides를 분리하였다. 3개 anti-fungal peptides는 분자량이 4-16 kDa 사이의 분명한 구별이 있을 뿐만 아니라, 각 peptide는 anti-fungal peptides 작용이 있었다. 이들 peptide의 공통 특징은 모두 열을 받은 뒤 활성이 변하지 않는 비교적 강한 내열성을 보여주었다.

Upregulation of Kruppel-like Factor 4 Gene expression by Allomyrina dichotoma Hemolymph in the INS-1 Pancreatic β-cells

Kisang Kwon,Hyun-Woo Suh,Hong Geun Kim,O-Yu Kwon 대한의생명과학회 2020 Biomedical Science Letters Vol.26 No.1

The hemolymph of Korean rhinoceros Allomyrina dichotoma consists of blood and lymph in which various kinds of proteins function physiologically. We have previously demonstrated that A. dichotoma hemolymph has the potential to treatment and prevent diabetes through activating transcription factor 3-gene (ATF3) regulation. In this study, we investigate the expression of Kruppel-like factor 4 (KLF4) in A. dichotoma hemolymph-treated INS-1 pancreatic β-cells. The new findings show that A. dichotoma hemolymph, which upregulates KLF4 gene expression in a dose-dependent and timedependent manner. In addition, hemolymph combine with mild endoplasmic reticulum (ER) stress, which also differentially regulates KLF4 gene expression. These results may provide insights to KLF4 gene-related disease therapies through KLF4 gene regulation.

Lysozyme as Pathogen-Recognition Protein in the Hemolymph of Galleria mellonella

Lee, In-Hee 한국곤충학회 2003 Entomological Research Vol.33 No.3

Recognition of invading micro-organisms into hemolymph is a pivotal event for triggering diverse immune mechanisms in insects. It has been known that this recognition was mediated by the binding of hemolymph proteins to pattern-molecules on the cell surface of microbes. Recently, I found that the lysozyme in the G. mellonella hemolymph has binding affinity to cell-walls of Gram (-), (+) bacteria and fungus (Candida albicans). After the hemolymph was incubated with heat-killed microbes and treated with acidic buffer containing high concentration of NaCl, several plasma proteins detached from microbes were detected by reverse phase HPLC and SDS-PAGE analyses. Of binding proteins, it was assumed that the major one might be a lysozyme, which was previously characterized in the G. mellonella hemolymph. Furthermore immunoblot analysis performed with antiserum to G. mellonella lysozyme revealed that it was a lysozyme.

방사선 손상 Balb/C 마우스 모델에서 누에 체액(Silkworm Hemolymph)의 간조직 보호 효과

남유리,강정애,노종국,최미희,Hayu Tyas Utami,장범수,박상현 한국방사선산업학회 2014 방사선산업학회지 Vol.8 No.1

This study was designed to evaluate the protective effect of silkworm hemolymph againstγ-irradiation induced damage in the liver of mice. Female Balb/C mice (6 weeks old) were exposedto γ-irradiation (6 Gy) and administered orally to silkworm hemolymph (5 ml kg-1 BW) for 7 dayspost-irradiation. The body weight, spleen index, plasma aspartate transaminase (AST), plasmaalanine transaminase (ALT), and liver malondialdehyde (MDA) levels were determined. Comparedwith irradiated control mice, the activity of plasma AST and the level of MDA were significantlydecreased in mice treated silkworm hemolymph. These results show that silkworm hemolymph isfound to have a protective effect against γ-irradiation induced damage in mice.

韓國産 貝類의 Hemolymph에 대한 Disc-電氣泳動橡의 比較硏究(Ⅰ)

최흥민,우세홍,한양일,나동진 최신의학사 1974 最新醫學 Vol.17 No.3

韓國産 海産貝類 8種의 hemolymph에 대하여 disc-電氣泳動한 結果 개조개 9개, 꾜막7개, 바지락8개, 동죽6개, 홍합4개, 콩조개6개 남방재첩 9개. 대합7개의 泳動帶를 가진 泳動像을 얻을 수 있었고 各各의 特徵에 따른 差異點을 發見할 수 있었다. 이것이 더 나아가 外形을 分別할 수 없을 때의 種을 鑑別하여 낼 수 있는 하나의 factor가 될 수 있다. The identify of a Pelecipoda is usually readily determined from the physical appearance of the Pelecipoda. The present objective methods of identifying Pelecipoda is based on the species-specific protein separation patterns obtained on electrophoresis of the hemolymph of the Pelecipoda. This is also can be separated by electrophoresis into species characteristic patterns that could be used for species identification. The separation patterns obtained on polyacrylamide gel for the hemolymph of fresh Saxidomus purpuratus, Tagillarca granosa, Tapes philippinarum, Mactra venerifomis, Mytilus edulis, Corbicula felnouilliana, Coricula fluminea, Meretrix lusoria are presented.