两种多糖对日本沼虾酚氧化酶活力的影响
EFFECTS OF TWO KINDS OF POLYSACCHARIDE ON THE PHENOLOXIDASE ACTIVITY IN MACROBRACHIUM NIPPONENSE
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摘要: 在饲料中分别添加1‰的云芝多糖和虫草多糖,对日本沼虾进行投喂,通过连续测定日本沼虾血清中的酚氧化酶活力,研究两种多糖对日本沼虾免疫机能的作用.结果表明,在平均水温为23.8℃±3.8℃时,云芝组、虫草组分别较对照组提高了3.52%、8.62%;在平均水温为10.8±2.℃时,云芝组和虫草组分别较对照组提高了30.02%、5.12%,因而云芝多糖和虫草多糖能明显地增强日本沼虾血清的酚氧化酶活力.另经实验表明,在王雷所确立的酚氧化酶活力的检测条件下,其PO活力为8.3units/mL,而在Horowitz所确立的检测条件下,其PO活力为290units/mL,并且后者检测方法重复性好,能较好地反映日本沼虾的免疫状态.Abstract: Macrobrachium nipponense,2.5-3g in body weight, was bought from aquaculture market in Wuhan, and then cultured in laboratory tank (65cm×45cm×34cm). The shrimps were kept in the tank for 7 days and fed with diet containing 1‰ Polysticus and Cordyceps polysaccharide. The haemolymph was then taken from the heart of M.nipponense. which were cultured for 1,4,7,14,21,28 days to determine the phenoloxidase (PO) activity. The results were as follows: 1 Acording to Wang Lei's method, the PO activity was 8.3 units/mL, while the PO activity was 290 units/mL by the Horowitz's method. Besides the high sensitivity, the latter had a better repetition than the former. Therefore, the Horowitz's method was better for responding the immune state of M.nipponense. 2. PO activity could be tested in the haemolymph of M.nipponense. There was little effect on the PO activity by adding SDS. It was clear that in the haemolymph PO was not present by the form of zymogen. 3. At high temperature (for example 23.8±3.8℃), M.nipponense had a vigorously ingestion, the PO activity of Polystictus and Cordyceps groups increased about 34.52% and 84.62%. Polystictus and Cordyceps polysaccharide had remarkable enhancement on the PO activity. While the increase of PO activity was 30.02% and 5.12%respectively in low water temperature. The enhancement of polysaccharide on the PO activity was not significant. The increase of PO activity indicated that Polystictus and Cordyceps polysaccharide could improve nonspecific immunity of the shrimps, which would be useful as a potential immunostimulant.
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[1] Sderhll K, Cerenius L Johansson M W. The prophenoloxidase activating system and its role in invertebrate defense [J]. Ann. N. Y. Acad. Sci. 1994, 12: 155-161[2] Sderhll K, Cerenius L. Role of the prophenoloxidase-activating system in invertebrate immunity [J]. Curr. Opin. Immunol. 1998,10:23-28[3] Thornqvist PO, Johansson MW, Soderhall K. Opsonic activity of cell adhesion proteins and beta-1, 3-glucan binding proteins from two crustaceans [J]. Dev Comp Immunol. 1994, 18(1):3-12[4] Meng FL, Zhang Y Z, Kong J. The research review of prophenoloxidase activating systems in crustacean [J]. Oceanolgia et Limnologia Sinica, 1999,30(1):110-115.[孟凡伦,张玉臻,孔健,等.甲壳动物的酚氧化酶原激活系统的研究评价[J].海洋与湖沼, 1999,30(1):110-115][5] Wang L, Li G Y, Mao Y X. Measuring methods and variations of some haemolymph factors in penaeus chinensis after their oral ingestion of immuno drugs [J]. Oceanolgia et Limnologia Sinica. 1995,26(1):34-41.[王雷,李光友,毛远兴.口服免疫药物后中国对虾某些血淋巴因子的测定及方法研究[J].海洋与湖沼, 1995,26(1):34-41][6] Luo R X. Induction of immunity substance in Penaeus chinensis by Chinese herbal medcine. Oceanolgia et Limnologia Sinica [J]. 1997,28(6):573-578.[罗日祥.中药制剂对中国对虾免疫活性物的诱导作用[J].1997,28(6):573-578][7] Li T D, Yu J, Yu K K. Studies on the phenoloxidase in the serum of penaeus chinensis [J]. Transactions of Oceanology and Limnology. 1998,1:51-55.[李天道,于佳,俞开康. 中国对虾血清中酚氧化酶活力的研究[J]. 海洋湖沼通报, 1998,1:51-55][8] Aspán A, Huang T S. Cerenius L, Sderhll, K. cDNA cloning of prophenoloxidase from the freshwater crayfish Pacifastacus leniusculus and its activation [J]. Proc. Natl. Acad. Sci. USA 1995, 92, 939-943[9] Horowitz NH, Shen S. Neurospora tyrosinae[J]. J, Boil Chem, 1952, 197:573-520[10] Saul S J. The majority of prophenoloxidase in the hemolymph of Manduca sexta is present in the plasma and not in the hemocytes [J]. Dev Comp Immune, 1985,11:479-485[11] Wang L, Li G Y,Mao Y X. Studies on the activities and characteristics of the antibacterial, bacteriolysis and phenoloxidase in the haemolymph of Penaeus chinensis [J]. Oceanolgia et Limnologia Sinica, 1995,26(2):179-185.[王雷,李光友,毛远兴.中国对虾血淋巴中的抗菌、溶菌活力与酚氧化酶活力的测定及其特性研究.海洋与湖沼,1995,26(2):179-185][12] Lee SY, Wang R, Soderhall K. A lipopolysaccharide- and beta-1,3-glucan-binding protein from hemocytes of the freshwater crayfish Pacifastacus leniusculus. Purification, characterization, and cDNA cloning [J]. J Biol Chem. 2000, 275(2):1337-1343［13］Soderhall K, Hall L.Lipopolysaccharide induced activation of the prophenoloxidase activating system in crayfish haemocyte[J]. Biochem Biophys Acta, 1984, 797:99-104 Sderhll K, Cerenius L Johansson M W. The prophenoloxidase activating system and its role in invertebrate defense [J]. Ann. N. Y. Acad. Sci. 1994, 12: 155-161[2] Sderhll K, Cerenius L. Role of the prophenoloxidase-activating system in invertebrate immunity [J]. Curr. Opin. Immunol. 1998,10:23-28[3] Thornqvist PO, Johansson MW, Soderhall K. Opsonic activity of cell adhesion proteins and beta-1, 3-glucan binding proteins from two crustaceans [J]. Dev Comp Immunol. 1994, 18(1):3-12[4] Meng FL, Zhang Y Z, Kong J. The research review of prophenoloxidase activating systems in crustacean [J]. Oceanolgia et Limnologia Sinica, 1999,30(1):110-115.[孟凡伦,张玉臻,孔健,等.甲壳动物的酚氧化酶原激活系统的研究评价[J].海洋与湖沼, 1999,30(1):110-115][5] Wang L, Li G Y, Mao Y X. Measuring methods and variations of some haemolymph factors in penaeus chinensis after their oral ingestion of immuno drugs [J]. Oceanolgia et Limnologia Sinica. 1995,26(1):34-41.[王雷,李光友,毛远兴.口服免疫药物后中国对虾某些血淋巴因子的测定及方法研究[J].海洋与湖沼, 1995,26(1):34-41][6] Luo R X. Induction of immunity substance in Penaeus chinensis by Chinese herbal medcine. Oceanolgia et Limnologia Sinica [J]. 1997,28(6):573-578.[罗日祥.中药制剂对中国对虾免疫活性物的诱导作用[J].1997,28(6):573-578][7] Li T D, Yu J, Yu K K. Studies on the phenoloxidase in the serum of penaeus chinensis [J]. Transactions of Oceanology and Limnology. 1998,1:51-55.[李天道,于佳,俞开康. 中国对虾血清中酚氧化酶活力的研究[J]. 海洋湖沼通报, 1998,1:51-55][8] Aspán A, Huang T S. Cerenius L, Sderhll, K. cDNA cloning of prophenoloxidase from the freshwater crayfish Pacifastacus leniusculus and its activation [J]. Proc. Natl. Acad. Sci. USA 1995, 92, 939-943[9] Horowitz NH, Shen S. Neurospora tyrosinae[J]. J, Boil Chem, 1952, 197:573-520[10] Saul S J. The majority of prophenoloxidase in the hemolymph of Manduca sexta is present in the plasma and not in the hemocytes [J]. Dev Comp Immune, 1985,11:479-485[11] Wang L, Li G Y,Mao Y X. Studies on the activities and characteristics of the antibacterial, bacteriolysis and phenoloxidase in the haemolymph of Penaeus chinensis [J]. Oceanolgia et Limnologia Sinica, 1995,26(2):179-185.[王雷,李光友,毛远兴.中国对虾血淋巴中的抗菌、溶菌活力与酚氧化酶活力的测定及其特性研究.海洋与湖沼,1995,26(2):179-185][12] Lee SY, Wang R, Soderhall K. A lipopolysaccharide- and beta-1,3-glucan-binding protein from hemocytes of the freshwater crayfish Pacifastacus leniusculus. Purification, characterization, and cDNA cloning [J]. J Biol Chem. 2000, 275(2):1337-1343［13］Soderhall K, Hall L.Lipopolysaccharide induced activation of the prophenoloxidase activating system in crayfish haemocyte[J]. Biochem Biophys Acta, 1984, 797:99-104
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