摘要
贯叶连翘(Hypericum perforatum)是藤黄科(Guttiferae)金丝桃属(Hypericum)多年生草本植物,是一种传统草药,广泛应用于欧洲,西亚和北非。自古以来,贯叶连翘通常用来抗抑郁症。然而贯叶连翘含有的多种成分还具有抗病毒、抗菌、抗氧化等活性。特别是最近一些研究表明贯叶连翘可以降低动物血清中甘油三酯和总胆固醇含量。国内外将贯叶连翘作为动物饲料添加剂的研究并不多见,生理生化和免疫功能等方面的研究少有报道。
本研究将A、B、C、D、E五个添加水平(A<D<C<D<E)的贯叶连翘提取物添加于鸡饲料中。经过一段时间的饲喂,定期检测血液和鸡蛋中的一系列生理生化指标,考察贯叶连翘对鸡血液生理、血清生化、生产性能、鸡蛋品质、鸡蛋生化指标、蛋清溶菌酶等相关指标的影响。另外,在首次和第二次H5亚型禽流感疫苗免疫后的7天内,将A、C、D、E四个添加水平的贯叶连翘提取物添加于鸡饲料中,定期检测H5亚型禽流感疫苗免疫效果.最终,筛选出最佳的贯叶连翘添加比例和饲喂方法,以期提高鸡的生产性能和防治禽流感能力,为贯叶连翘饲料添加剂的生产应用提供理论依据。
主要研究结果如下:
1、D添加水平的两组鸡血清中白蛋白、谷丙转氨酶、碱性磷酸酶等肝功能指标和血肌酐与对照组相比无显著差异(P>0.05),血液中白细胞数和淋巴细胞百分率明显提高(P<0.05),从而提高了非特异性免疫能力。另外,鸡血清溶菌酶活力明显提高(P<0.05),从而增强了抵御病菌感染的能力。
2、饲喂贯叶连翘4周后,与对照组相比,E添加水平增加了(P<0.05)蛋重,降低了(P<0.05)料蛋比(耗料重/蛋重)和蛋壳厚度;D和E添加水平,降低(P<0.05)蛋黄总胆固醇和丙二醛(MDA)含量,提高(P<0.05)蛋清溶菌酶活力,一定程度上提高(P>0.05)超氧化物歧化酶(SOD)活力,但无显著性差异。D、E两个添加水平对产蛋率都无影响(P>0.05)。此外,B、C、D三个添加水平都显著降低(P<0.05)蛋黄甘油三酸脂含量。研究结果表明,在饲喂贯叶连翘4周时间内,除E添加水平能使蛋壳厚度降低之外,其它各添加水平均未引起蛋品质的降低。
3、D水平添加量的贯叶连翘能使鸡血清中H5亚型禽流感病毒抗体水平在首免和二免后的抗体高峰期提高9.82%、30.63%;使H5亚型禽流感病毒Re-4株抗体效价在首免和二免后的抗体高峰期提高11.27%、27.40%;使H5亚型禽流感病毒Re-5株抗体效价在首免和二免后一直高于对照组,特别是在首次、二次免疫后的抗体高峰期分别提高7.70%、16.44%。
综上所述,在D添加水平下,贯叶连翘对鸡的生理功能、生产性能和鸡蛋品质无不良影响,提高鸡的非特异性免疫作用和鸡蛋的氧化稳定性,同时降低蛋黄总胆固醇和甘油三酯含量;另外,D水平添加量的贯叶连翘能够有效增强H5亚型禽流感疫苗的免疫效果,维持首次免疫和二次免疫后的抗体水平长期高于对照组。因此,可以得出结论,贯叶连翘在鸡饲料中最佳的添加水平是D;另外,贯叶连翘能够在保证鸡生理安全的前提下,提高蛋鸡生产性能和鸡防治H5亚型禽流感能力,提高鸡蛋抗氧化性能,降低鸡蛋中脂类含量,从而提高养殖场的经济效益,产出的鸡蛋也更加有益于人体健康。
Hypericum perforatum is a perennial herb in the Guttiferae family native to Europe, western Asia, and northern Africa. It has traditionally been applied as a medicinal plant to treat antidepressant. Currently, it has been found that various ingredients contained in Hypericum perforatum have anti-virus, anti-bacterial, anti-oxidation activities, especially can lower the serum total cholesterol and triglyceride contents in animals.
Inspite of those above, the study on appling Hypericum perforatum as feed additives for animals has not been reported both at home and abroad. The study was few on the effects of Hypericum perforatum on physiological and immune actions in layers. In this study, several levels (A<D<C<D<E) of Hypericum perforatum extract were added to the layers'diets. Afterthat, the effects of it were studied on physiological and biochemical indexes in blood, production performances and the level of antibody to H5 subtype avian influenza in layers. Finally, the best level of Hypericum perforatum added into diets was filtered out, in order to apply it as layers'additive and enhance the production performances and ability to prevent avian influenza. The results are as following:
1. The contents of albumin, alanine aminotransferase, and alkaline phosphatase in serum of D level groups had no significant difference (P>0.05) compared with the control group. White blood cell count and lymphocyte percentage significantly increased (P<0.05) in the two level groups, thus Hypericum perforatum can enhance the non-specific immunity. In addition, the lysozyme activity in serum was significantly increased (P<0.05), increasing the ability to resist infection.
2. After feding with Hypericum perforatum for 4 weeks, compared with the control group, E level had increased (P<0.05) egg weight, reduced (P<0.05) feed conversion ratio (feed consumption weight/egg weight) and egg shell thickness. D and E levels had decreased (P<0.05)total cholesterol and malondialdehyde (MDA) content egg yolk, increased (P<0.05) egg white lysozyme, improve superoxide dismutase (SOD) activity to some extent, but with no significant difference (P>0.05). D and E levels have no effect on egg production rate (P>0.05). In addition, B, C and D levels significantly lowered (P<0.05) the content of triglycerides egg yolk. The results also showed that the egg qualities were not influenced during 4 weeks feeding except that the E level can reduce the thickness of eggshell.
3. D additional level of Hypericum perforatum can increased the level of antibody to H5 subtype avian influenza in serum by 9.82% and 30.63% respectively in the two antibody peaks after first and second immunizations. It can increased Re-4 polyclonal H5 subtype avian influenza antibody titer in serum by 11.27% and 27.40% respectively in the two antibody peaks after first and second immunizations. Re-5 polyclonal H5 subtype avian influenza antibody titer in D level group were always higher than control after the twice immunizations, particularly the antibody titer was increased by 7.70% and 16.44% respectively in antibody peaks.
To sum up, D level of Hypericum perforatum added into diets had no adverse effects on the physiological function of layers and egg qualitys. It can enhance the non-specific immunity, production performance of layers and egg oxidation resistance,while it can reduced total cholesterol and triglyceride contents in egg yolks. In addition, D level of Hypericum perforatum added into diets can effectively enhance the action of H5 subtype avian influenza vaccine with higher antibody levels after the first and secondary immunization. Therefore, it can be concluded that the best level of Hypericum perforatum added into diets is D. Moreover, Hypericum perforatum can improve the production performance of layers, the prevention against H5 subtype avian influenza and the oxidation resistance of eggs, while reduce lipid content of eggs, without affecting the physical safety of layers. In a word, the Hypericum perforatum may enhance the profits of the farm, and be beneficial to human health.
引文
[1]中国科学院中国植物志编辑委员会.中国植物志(50卷第二分册)[M].北京:科学出版社,1990.
[2]Blumenthal M, Busse W R, Goldberg A, et al. The complete German Commission E monographs:therapeutic guide to herbal medicines[M]. Boston:American Botanical Council,1998.
[3]中科院植物研究所.中国高等植物图鉴[M].第二册.北京:科学出版社,1985:875.
[4]刘兵.贯叶连翘研究进展-原植物采收、制剂和化学成分[J].国外医药(植物学分册),1998,13(3):99-103.
[5]中科院植物研究所.中国高等植物图鉴[M].北京:科学出版社,1972:879.
[6]Tatsis E C, Boeren S, Exarchou V, et al. Identification of the major constituents of Hypericum perforatum by LC/SPE/NMR and/or LC/MS[J]. Phytochemistry,2007,68(3): 383-393.
[7]Lu H F, Shen Z G, Li J Y, et al. The patterns of secretory structure and their relation to hypericin content in Hypericum[J]. Acta Bot. Sin.2001,43:1085-1088.
[8]胡正海,吕洪飞,李景原.金丝桃属植物的分泌结构与金丝桃素关系的研究[J].西北大学学报(自然科学版),2002(5):459-464.
[9]郁青,沈兆邦,王成章,等.贯叶金丝桃(贯叶连翘)研究进展[J].林产化工通讯,2000,34(1):22-27.
[10]梁小燕.金丝桃属植物的研究进展[J].广西植物,1998,18(3):256-262.
[11]吕洪飞,胡正海.贯叶连翘分泌结构的发育及其内含物积累的研究[J].西北植物学报,2001,21(2):287-292.
[12]郑清明.金丝桃属药用植物的生药学研究[J].中西医结合学报,2004,2(3).
[13]张文龙,王秋芳,丛日华,等.中草药添加剂对肉鸡胆固醇代谢的影响[J].中国兽医科技,2004,34(6):61-64.
[14]Lawvere S, Mahoney M C. St. John's wort[J]. Am Fam Physician,2005(72):2249-2254.
[15]Miller A L. St. John's wort (Hypericum perforatum):clinical effects on depression and other conditions[J]. Altern Med Rev,1998(3):18-26.
[16]张筱,卢定强,权静,等.贯叶连翘抗抑郁研究新进展[J].生物加工过程,2004,2(1):11-15.
[17]Kirakosyan A, Sirvent T M, Gibson D M, et al. The production of hypericins and hyperforin by in vitro cultures of St. John's wort (Hypericum perforatum)[J]. Appl Biochem,2004(39):71-81.
[18]朴香兰,王雷,朴香淑,等.连翘提取物对肉仔鸡热应激下生长性能和抗氧化活性的影响[J].中国畜牧杂志,2008,44(3):23-26.
[19]杨得坡,甘良春,胡海燕.国外对贯叶连翘抗抑郁疗效的临床验证[J].中西医结报,2004,2(3):231-238.
[20]Lavie G F, Valentine, Levin B, et al. Studies of the mechanisms of action of the antiretroviral agents hypericin and pseudohypericin[J]. Proc Natl Acad Sci USA,1989,86: 5963-5967.
[21]Schempp C M, Windeck T, Hezel S, et al. Topical treatment of atopic dermatitis with St. John's wort cream-a randomized, placebo controlled, double blind half-side comparison[J]. Phytomedicine,2003,10 Suppl 4:31-37.
[22]李宏,邹国林.贯叶连翘总提取物对谷氨酸棒状杆菌和粪产碱杆菌的抗菌作用[J].武汉植物学研究,2002,20(3):213-218.
[23]Benedi J, Arroyo R, Romero C, et al. Antioxidant properties and protective effects of a standardized extract of Hypericum perforatum on hydrogen peroxide-induced oxidative damage in PC12 cells[J]. Life Sci,2004(75):1263-1276.
[24]Niko Radulovic, Vesna Stankov-Jovanovic, Gordana Stojanovic, et al. Screening of in vitro antimicrobial and antioxidant activity of nine Hypericum species from the Balkans[J]. Food Chemistry,2007,103(1):15-21.
[25]Bruno A Silva, Malva Joao O, Alberto C, et al. St. John's Wort(Hypericum perforatum) extracts and isolated phenolic compounds are effective antioxidants in several in vitro models of oxidative stress[J]. Food Chemistry,2008,110(3):611-619.
[26]Michael Spiteller, Tevfik Ozen, Andrija Smelcerovic, et al. Phenolic constituents and the in vitro antioxidant activity of the flowers of Hypericum venustum[J]. Fitoterapia,2008, 3(79):191-193.
[27]Juana Benedi, Rocio Arroyo, Carmen Romero, et al. Antioxidant properties and protective effects of a standardized extract of Hypericum perforatum on hydrogen peroxide-induced oxidative damage in PC12 cells[J]. Life Sciences,2004,75(10):1263-1276.
[28]Doaa A. El-Sherbiny, Khalifa Amani E, Attia Amina S, et al. Hypericum perforatum extract demonstrates antioxidant properties against elevated rat brain oxidative status induced by amnestic dose of scopolamine[J]. Pharmacology Biochemistry and Behavior, 2003,3-4(76):525-533.
[29]Fidan Hakimoglu, Goksel Kizil, Zeki Kanay, et al. The effect of ethanol extract of Hypericum lysimachioides on lipid profile in hypercholesterolemic rabbits and its in vitro antioxidant activity[J]. Atherosclerosis,2007,1(192):113-122.
[30]Zou Y, Lu Y, Wei D. Hypocholesterolemic effects of a flavonoid-rich extract of Hypericum perforatum L. in rats fed a cholesterol-rich diet[J]. J Agric Food Chem,2005, 53:2462-2466.
[31]Chung D J, Kim H Y, Park K H, et al. Park Black cohosh and St. John's wort (GYNO-Plus) for climacteric symptoms[J]. Yonsei Med J,2007,48:289-294.
[32]Carlson, L. A. Serum lipids and atherosclerotic disease[M]//Carlson L A, Pernow B. Metabolic Risk Factors in Ischemia CardioVascular Disease. New York:Raven Press, 1982:1-6.
[33]Pekkanen A N, Punsar S, Karvonen M J. Short and long term association of serum cholesterol with mortality[J]. Am J Epidemiol,1992,135:1252-1258.
[34]American Heart Association. Dietary guidelines for healthy American adults[J]. Circulation,1986,74:1465a-1468a.
[35]赵咏梅,李发荣,杨建雄,等.连翘苷降血脂及抗氧化作用的实验研究[J].天然产物研究与开发,2005,17(2):157-159.
[36]刘兴吉,鞠砚,孟艳,等.光活化的金丝桃素诱导胶质瘤细胞凋亡的实验研究[J].中国实验诊断学,2001,5(1):29-30.
[37]Evstifeeva T A, Sibiriak S V. Immunotropic properties of biologically active products obtained from John's wort (H. perforatum) [J]. Eksper Klin Form Log,1996,59 (1): 51-54.
[38]朱健如,沈更新,王晓团.贯叶连翘总提取物延缓衰老作用的研究[J].华中科技大学 学报(医学版),2002,31(6):659-611.
[39]Lenard J, Rabsona A, Vanderoef R. Photodynamic inactivation of infectivity of human immunodeficiency virus and other enveloped viruses using hypericin and rose bengal: inhibition of fusion and syncytia formation[J]. Proc Natl Acad Sci USA,1993,90 (1):158-162.
[40]Chen B, Roskans T, Xu Y, et al. Photodynamic therapy with hypericin induces vascular damage and apoptosis in the RIF-1 mouse tumor model[J]. Int J Cancer,2002, 98:284-290.
[41]Evstifeeva V A, Sibiryak S V. The immunotropic properties of biologically active products obtained from Klamath weed (Hypericum perforatum L.) [J]. Eksp Klin Farmakol,1996,59:51-54.
[42]娄海容,靳士英.在防治禽流感中引人注目的中草药——金丝桃属和金丝桃素[J].现代医院,2006(16):3-5.
[43]Prince A M, Pascual D, Meruelo D, et al. Strategies for evaluation of enveloped virus inactivation in red cell concentrates using Hypericin[J]. Photochem Photobiol,2000,71 (2):188-195.
[44]尚若峰,梁建平.防治禽流感及其他RNA病毒的新兽药-金丝桃素[J].兽药与安全,2005,10:42.
[45]郭腾,吴连勇.金丝桃素的最新研究进展[J].兽药与饲料添加剂,2006(11):7-9.
[46]时玉声,崔中林.兽医临诊检验手册[M]..上海:上海科学技术出版社,1989,159-167.
[47]瞿自明.兽医中草药大全[M].北京:中国农业科技出版社,1989,147-149.
[48]Thomas, C, R. S. Macgill, G. C. Miler, et al. Photoactivation of hypericin generates singlet oxygen in mitochondria and inhibits succinoxidase[J]. Photochem Photobiol,1992, 55 (1):47-53.
[49]Vanberweghem J. L., M. Depierrcux, et al. Rapidly progressive intersitial fibrosis in young women:association with slimming regimen including Chinese herbs[J]. Lancet, 1993,341:387.
[50]Zhao R Q, Decuypere E, Grossmann R. Secretory patterns GH and IGF- Iconcentrations in meat-type and laying-type chickens during early post hatch development[J]. Poul Avian Bio Rev,1996,6:270-271.
[51]Wenk C. Rencent advances in animal feed additives such as metabolic modifiers, antimicrobial agents, probiotics, enzymes and highly available minerals[J]. J Anim Sci, 2000,13(1):86-95.
[52]丁美方,王有月,魏秀莲.浅谈中草药饲料添加剂[J].饲料研究,2007(3):14-16.
[53]姚峰,张存智,张见深,等.中草药饲料添加剂对肉用种鸡产蛋初期的影响[J].畜禽养殖,2007(6):36-37.
[54]Soliva C R, Widmer S and Kreuzer M. Ruminal fermentation of mixed diets supplemented with St. John's Wort (Hypericum perforatum) flowers and pine (Pinus mugo) oil or mixtures containing these preparations[J]. Journal of Animal and Feed Sciences, 2008,17:352-362.
[55]National Research Council. Nutrient Requirements of Poultry[M].9th rev. ed. Washington, DC:Natl. Acad. Press,1994.
[56]Papakostas G I, Crawford C M, Scalia M J, et al. Timing of clinical improvement and symptom resolution in the treatment of major depression disorder. A replication of findings with the use of a double-blind, placebo-controlled trial of Hypericum perforatum versus fluoxetine[J]. Neuropsychobiology 2007,56(2-3):132-137.
[57]卢宋藩.家禽及实验动物生理生化参数[M]北京:农业出版社,1983:95,160,213-214.
[58]冷超,韩凌霞,于海波,等.不同周龄BWEL-SPF种鸡生理生化指标的测定.中国比较医学杂志.2007,17(12):697-701.
[59]Shipochliev T, Dimitrov A, Aleksandrova E. Anti-inflammatory action of a group of plant extracts[J]. Vet Med Nauki,1981,18(6):87-94.
[60]傅芃,张川,张卫东.咕吨酮类化合物的药理活性研究进展[J].药学实践杂志,2005,23(1):5-7.
[61]Davisl A K, Maney D L, Maerz J C. The use of leukocyte profiles to measure stress in vertebrates:a review for ecologists[J]. Functional Ecology, (2004).22, (5):760-772.
[62]Wayland M, Gilchrist H G, Marchant T, et al. Immune function, stress response, and body condition in arctic-breeding common eiders in relation to cadmium, mercury, and selenium concentrations [J]. Environ Res,2002,90(1):47-60.
[63]崔耀明.兽医基础[M].郑州:河南科学技术出版社,2007.
[64]Dost T, Ozkayran H, Gokalp F, et al. The Effect of Hypericum perforatum (St. John's Wort) on Experimental Colitis in Rat[J]. Dig Dis Sci,2009,54(6):1214-1221.
[65]Shibayama Y, Kawachi A, et al. Effect of pre-treatment with St John's Wort on nephrotoxicity of cisplatin in rats[J]. Life Sci,2007,81(2):103-108.
[66]康格非.临床生物化学和生物化学试验(第2版)[M].北京:人民卫生出版社,1999:169.
[67]杨汉春.动物免疫学[M].北京:中国农业大学出版社[M],2003:32-33
[68]Verotta L, Lovaglio E, Sterner O, et al. Modulation of chemoselectivity by protein additives. Remarkable effects in the oxidation of hyperforin[J]. J Org Chem,2004:69(23): 7869-7874.
[69]黄承乐.十一种生化项目在肝脏疾病中变化的研究[J].右江民族医学院学报,199921(5):763-765.
[70]Avenirova E L. Effect of novoimanine on the cellular permeability indices of staphylococci[J]. Antibiotiki,1977,22(7):630-634.
[71]贾向志,李元.溶菌酶的研究进展[J].生物技术通讯,2002,3(5):374-377.
[72]Derbentse N A, Rabinovych A S, Aizenman B, et al. Antimicrobial substances from Hypericum perforatum[J].Mikrobiol Zh,1959,21(5):52-57.
[73]Zakharova N S, Remova T N, Shepeleva I B, et al. Action of plant extracts on the natural immunity indices of animals[J]. Zh Mikrobiol Epidemiol Immunobiol,1986, (4):71-5.
[74]Schempp C M, Pelz K, Wittmer A, et al. Antibacterial activity of hyperforin from St John's wort, against multiresistant Staphylococcus aureus and Gram-positive bacteria[J]. Lancet,1999,353:2129.
[75]Reichling J, Weseler A, Saller R. A current review of the antimicrobial activity of Hypericum perforatum L[J]. Pharmacopsychiatry,2001,34 Suppl 1:S116-118.
[76]Bai, W. B., Y. M. Wei, and Y. S. Guo. Study on the extraction and purification of hypericin from hypericum perforatum L. with ethanol[J]. Journal of traditional Chinese veterinary medicine.2005,24(3):17-18.
[77]Li, J., Q. Mo, J. Zhang, and F. Li.1999. Study on extract of hypericin from hypericum perforatum L. with ovthogonal experimental design [J]. Chemical research and application 11(1):99-101.
[78]李志琼.α-亚麻酸对产蛋鸡脂质代谢及蛋黄胆固醇沉积的影响及其机理[D].四川农业大学,2007.
[79]郝志明.鸡蛋、鸡胚的SOD和MDA水平[J].西北联合大学学报,2004,7(5):11-13.
[80]潘学燕,高玉鹏.n-3系列多不饱和脂肪酸(PUFA)富集保健功能蛋开发研究[J].西北农业大学学报,2008,17(1):20-26.
[81]Kascakova S, Nadova Z, Mateasik A, et al. High level of low-density lipoprotein receptors enhance hypericin uptake by U-87 MG cells in the presence of LDL[J]. Photochem Photobiol,2008,84(1):120-127.
[82]李敏芳,李慧,王学美.金丝桃苷药理作用研究进展[J].中国中医药信息杂志,200815(4):102-104.
[83]李治学,魏丽娜,章世元.鸡蛋壳质量与超微结构关系的研究[J].中国畜牧杂志,2008,1:35-39.
[84]Krusekopf S, Roots I. St. John's wort and its constituent hyperforin concordantly regulate expression of genes encoding enzymes involved in basic cellular pathways[J]. Pharmacogenet. Genomics,2005,15:817-829.
[85]Breyer A, Elstner M, Gillessen T, et al. Glutamate-induced cell death in neuronal HT22 cells is attenuated by extracts from St. John's wort(Hypericum perforatum L.)[J]. Phytomedicine,2007,14:250-255.
[86]Gilani A H, Khan A U, Subhan F, et al. Antispasmodic and bronchodilator activities of St. John's wort are putatively mediated through dual inhibition of calcium influx and phosphodiesterase[J]. Fundam Clin Pharmacol,2005,19(6):695-705.
[87]Silva B, Oliveira P J, Dias A, et al.. Quercetin, kaempferol and biapigenin from Hypericum perforatum are neuroprotective against excitotoxic insults[J]. Neurotox Res, 2008,13:265-279.
[88]Marsh W L, Davies J A. The involvement of sodium and calcium ions in the release of amino acid neurotransmitters from mouse cortical slices elicited by hyperforin[J]. Life Sci, 2002,71:2645-2655.
[89]王人悦.绿茶粉及茶多酚对蛋鸡血液生化指标、生产性能及鸡蛋品质的影响[D].安徽农业大学,2007.
[90]Bebout D E, Hempleman S C. Calcium deficient diet, acetazolamide and gas exchange characteristics of avian eggshells[J]. Respir Physiol,1994,95:11-20.
[91]Hickey J J, Anderson D W. Chlorinated hydrocarbons and eggshell changes in raptorial and fish-eating birds[J]. Science,1968,162:271-273.
[92]Brian, Haveson. Herbal appetite suppressant and weight loss composition[P]. HPF, L.L.C., assignee. US Pat. No.5,798,101.1998.
[93]王学静,臧素敏,张丽,等.银杏叶提取物对肉仔鸡生长性能、血脂和抗氧化功能的影响[J].畜牧与兽医,2006,38(11):40-42.
[94]徐皓.贯叶连翘的化学成分及药理作用研究[J].安徽农业科学,2007,35(14):4219-4221.
[95]Conforti F, Statti G A, Tundis R, et al. Antioxidant activity of methanolic extract of Hypericum triquetrifolium Turra aerial part[J]. Fitoterapia,2002(73):479-483.
[96]Gianetti J, Pedrinelli R, Petrucci R, et al. Inverse association between carotid intima-media thickness and the antioxidant lycopene in atherosclerosis[J]. Am Heart J, 2002,143(3):467-474.
[97]E1-Midaoui A, Champlain J de. Prevention of hypertension, insulin resistance, and oxidative stress by alpha-lipoic acid[J]. Hypertension,2002(39):303-307.
[98]Yakobson M G., Antonov A R, Golovatyuk A V, et al. Iron content and parameters of blood antioxidant activity in rats with hereditary arterial hypertension during experimental myocardial infarction[J]. Bull Exp Biol Med,2001,132 (5):1041-1044.
[99]Fang J L, Vaca C E, Valsta L M, et al. Determination of DNA adducts of malonaldehyde in humans:effects of dietary fatty acid composition[J]. Carcinogenesis,1996,17(5): 1035-1040.
[100]Sanchez-Reus M I, Gomez del Rio M A, Iglesias I, et al. Standardized Hypericum perforatum reduces oxidative stress and increases gene expression of antioxidant enzymes on rotenone-exposed rats[J]. Neuropharmacology,2007(52):606-616.
[101]Sloley B D, Urichuk L J, Ling L, et al. Chemical and pharmacological evaluation of Hypericum perforatum extracts[J].Acta Pharmacol Sin,2000(21):1145-1152.
[102]Singh A, Naidu P S, Gupta S, et al. Effect of natural and synthetic antioxidants in a mouse model of chronic fatigue syndrome[J]. J Med Food,2002(5):211-220.
[103]Doaa A El-Sherbiny, Khalifa Amani E, Attia Amina S, et al. Hypericum perforatum extract demonstrates antioxidant properties against elevated rat brain oxidative status induced by amnestic dose of scopolamine [J]. Pharmacology Biochemistry and Behavior, 2003,3-4(76):525-533.
[104]Berliner J A, Subbanagounder G, Leitinger N, et al. Evidence for a role of phospholipid oxidation products in atherogenesis[J]. Trends Cardiovasc. Med.2001(11):142-147.
[105]Thompson H J, Heimendinger J., A. Haegele, S. M. Sedlacek, C. Gillette, C. O'Neill, P. Wolfe, and C. Conry. Effect of increased vegetable and fruit consumption on markers of oxidative cellular damage[J]. Carcinogenesis,1999(20):2261-2266.
[106]高斌,高洪.氧自由基与细胞损伤[J].动物医学进展,2002,23(5):34-36.
[107]Sirvent T M, Krasnoff S B, and Gibson D M. Induction of hypericins and hyperforins in Hypericum perforatum in response to damage by herbivores[J]. J Chem Ecol,2003(29): 2667-2681.
[108]Gaind K N, Ganjoo T N. Antibacterial principle of Hypericum perforatum L.[J]. Indian. J.Pharm.,1959(21):172-175.
[109]Neuwald F, Hagenstrom U. Studies on antibacterial effect of Hypericum perforatum L.[J]. Arch. Pharm. Ber. Dtsch. Pharm. Ges.1954(287):439-441.
[110]Gurevich A I, Dobrynin V N, Kolosov M N, et al. Antibiotic hyperforin from Hypericum perforatum L[J]. Antibiotiki,1971(16):510-513.
[111]Bystrov N S, Chernov B K, Dobrynin V N, et al. The structure of hyperforin[J]. Tetrahedron. Lett.1975(32):2791-2794.
[112]Avato P, Raffo F, Guglielmi G, et al. Extracts from St. John's wort and their antimicrobial activity[J]. Phytother. Res.2004(18):230-232.
[113]Franklin G., Conceicao L F, Kombrink E, et al. Xanthone biosynthesis in Hypericum perforatum cells provides antioxidant and antimicrobial protection upon biotic stress[J]. Phytochemistry,2009(70):60-68.
[114]Vajs V, Vugdelija S, Trifunovic S, et al. Further degradation product of hyperforin from Hypericum perforatum (St John's Wort) [J]. Fitoterapia,2003(74):439-444.
[115]Havsteen B H. The biochemistry and medical significance of the flavonoids[J]. Pharmacol. Ther.,2002(96):67-202.
[116]Cunninghamal F E, Proctoral V A, Goetschal S J. Egg-white lysozyme as a food preservative:an overview[J]. Worlds Poult. Sci. J.1991 (47):141-163.
[117]国家标准化管理委员会.GB/T18936-2003.高致病性禽流感诊断技术[S].北京:中国标准出版社,2003.
[118]蒋建一,母安雄,王毅峰,等.不同禽流感疫苗免疫蛋鸡后抗体效价变化和消长规律研究[J].浙江农业科学,2004(4):221-222.