鹿角盘提取物抗炎作用的研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
鹿角盘有很高的药用价值,民间常用其来治疗乳腺炎、儿童腮腺炎和疮疖等。临床应用也表明鹿角盘具有良好的抗炎作用。但是目前还没有关于鹿角盘抗炎机理的研究报道,因此本论文拟通过炎症动物模型和细胞水平的研究来探讨鹿角盘提取物的抗炎功效及机理。
     1.采用二甲苯致小鼠耳肿胀模型评价相同生药量的鹿角盘微粉悬液(2.57 g/kg/d)、鹿角盘醇提物(0.12 g/kg/d)和鹿角盘水提物(0.44 g/kg/d)的抗炎效果,以氢化可的松(6mg/kg/d)为阳性对照(用药量皆参考《中国药典》)。正常小鼠灌胃给药十天后,给小鼠右耳涂抹二甲苯建小鼠耳肿胀模型。结果表明:微粉悬液组的肿胀率为132.46±15.26%,与空白对照组的肿胀率170.19±24.68%相比显著降低(P<0.05);鹿角盘水提物组和氢化可的松组的肿胀率分别为65.43±11.95%和64.63±11.73%,与空白对照组相比极显著降低(P<0.01);醇提物组肿胀率为154.03±22.43%,与空白对照组相比无显著差异。
     2.采用角叉菜胶致小鼠足跖肿胀模型来研究鹿角盘水提物的抗炎功效。正常小鼠灌胃给药十天后,给小鼠右后足注射角叉菜胶建小鼠足跖肿胀模型。结果表明:水提物高剂量组(0.44 g/kg/d)和氢化可的松组(6 mg/kg/d)小鼠的足跖肿胀率分别为43.68±2.32%、40.50±2.45%,与空白对照组(等体积生理盐水)小鼠的足跖肿胀率52.91±3.66%相比,均有极显著降低(P<0.01);水提物低剂量组(0.18 g/kg/d)小鼠的足跖肿胀率为48.46±2.22%,与空白对照组相比显著降低(P<0.05)。表明鹿角盘水提物对角叉菜胶引起的小鼠足跖肿胀有很好的消炎作用。
     检测小鼠血清和炎足浸出液中的炎症因子,表明鹿角盘水提物可以降低血清中NO、MDA的含量;降低炎足浸出液中PGE2含量;并增强血清中LZM(溶菌酶)、SOD(超氧化物歧化酶)和GSH-PX(谷胱甘肽过氧化物酶)的活力。因此,鹿角盘水提物的抗炎作用可能是:通过抑制NO的过量表达来减少细胞损伤;增强LZM的活力来改善小鼠体内的黏多糖代谢;增强SOD和GSH-PX的活力来减少脂质过氧化,从而减轻炎症反应。另外,鹿角盘水提物能够显著降低足跖肿胀小鼠炎足浸出液中IL-6的含量,而不影响IFN-y和IL-2的含量。说明鹿角盘水提物可能是通过抑制巨噬细胞相关炎症介质的表达来发挥抗炎作用的。
     3.将LPS诱导的RAW264.7细胞在鹿角盘水提物浓度为0-40 mg/mL的细胞培养液中培养12h,检测细胞培养液中NO、PGE2、IL-6的含量结果表明,随着鹿角盘水提物浓度的增加,细胞培养液中NO、PGE2、IL-6的含量逐渐降低,呈剂量依赖关系。
     研究表明,鹿角盘水提物有良好的抗炎作用,其抗炎作用可能是通过抑制机体炎症介质过量表达,抑制巨噬细胞相关细胞因子的表达和增强机体内相关酶的活力来实现的。
Antler Base has a high medicinal value. Over the past years people used it to treat mastitis, mumps and boils. Clinical applications have confirmed that Antler Base has sound anti-inflammatory effect. But there is no report for its anti-inflammatory mechanism. So this paper proposed to study the anti-inflammatory effect and its mechanism of Antler Base Extract by animal models and cell experiments.
     1. Xylene-induced ear edema mice model was used to evaluate the anti-inflammatory effect of Antler Base powder suspension(2.57 g/kg/d), ethanol extract of Antler Base(0.12 g/kg/d) and aqueous extract of Antler Base(0.44 g/kg/d), hydrocortisone(6 mg/kg/d) as a positive control. The results showed that Antler Base powder suspension group had lower swelling rate(132.46±15.26%) than the control(170.19±24.68%)(P<0.05). Aqueous extract group and hydrocortisone group had lower swelling rate (65.43±11.95%&64.63±11.73%) than the control (P<0.01). But ethanol extract didn't have this effect.
     2. Carrageenan-induced paw edema mice model is one of the most classical models of anti-inflammatory drug research. This model was used to study the anti-inflammatory effect of aqueous extract of Antler Base. The results showed that high-doses aqueous extract of Antler Base group (0.44 g/kg/d) and hydrocortisone group (6 mg/kg/d) had lower swelling rate (43.68±2.32%&40.50±2.45%) than the control 52.91±3.66%(P<0.01). Low-doses aqueous extract of Antler Base group (0.18 g/kg/d) had lower swelling rate (48.46±2.22%) than the control (P<0.05).
     Aqueous extract of Antler Base could also reduce the content of serum NO and MDA, reduce the content of PGE2 in leachate of inflammation foot and enhanc the activity of LZM, SOD and GSH-PX in serum. Its anti-inflammatory effects is probably by inhibiting the over-expression of NO to reduce cell trauma, enhancing LZM activity to improve the metabolism of polysaccharides, enhancing SOD and GSH-PX activity to reduce lipid peroxidation to reduce tissue damage and promote inflammatory tissue injury and recuperate. Aqueous extract of Antler Base could significantly reduce the content of IL-6 in leachate of inflammation foot, but it didn't affect IFN-y and IL-2 levels significantly. The results showed that aqueous extract of Antler Base might be playing anti-inflammatory effects by inhibiting monocyte-macrophage activation.
     3. Cultured·the LPS-induced RAW264.7 cells at 0-40 mg/mL concentration of aqueous extract of Antler Base for 12h, then analysis the content of NO,PGE2 and IL-6 in cell culture medium. The results showed that as the concentration of aqueous extract of Antler Base increase, the content of NO, PGE2 and IL-6 in cell culture medium decreased in a dose-dependent manner.
     The results show that aqueous extract of Antler Base has significant anti-inflammation effect. This effect may occur by inhabiting the body's over-expression of inflammatory mediators, inhabiting of macrophage-related cytokines and enhancing the activity of the related enzymes of body.
引文
[1]郭延蜀,郑慧珍.中国梅花鹿地史分布.中和亚种的划分及演化历史[J].兽类学报,2000,20(3):168-197.
    [2]盛和林.中国鹿科动物[M],上海:华东师范大学出版社,1992,202-212.
    [3]吴华,胡杰,方盛国等.中国圈养梅花鹿的遗传多样性和遗传结构[J].动物学杂志,2006,41(4):41-47.
    [4]张艳霞,孙冬梅.鹿源系列中药材的特点及应用[J].中国民间疗法,2010,18(9):61-62.
    [5]田玉华,胡薇.梅花鹿角脱盘化学成分的研究[J].畜牧与兽医,2010,42:235-236.
    [6]张金宝,试用鹿角盘治疗胃癌一例[J].畜牧兽医科技信息,2006,04:21.
    [7]王丽虹,高志光.鹿花盘水溶性成分的药理活性与临床应用[J].经济动物学报,1999,3(3):18-22.
    [8]陈玉山,王淑贤,王本祥.鹿花盘注射液的药理实验[J].特产研究,1989,4:9-12.
    [9]赵文静.复方鹿花盘胶囊对实验性乳腺增生的治疗研究[D].黑龙江:黑龙江中医药大学中药学,2008.
    [10]王志兵,邱芳萍,李志民等.鹿角盘活性成分对小鼠吞噬功能及对大鼠乳腺增生激素水平的影响[J].食品科学,2007,11:225-226.
    [11]王志兵,邱芳萍,解耸林.鹿角盘蛋白多肽的制备与活性研究[J].中国食品学报,2008,8(3):28-32.
    [12]苏凤艳,李慧萍,王艳梅等.鹿花盘蛋白质的提取与生物活性测定[J].动物科学与动物医学,2001,18(2):18.
    [13]史小青,刘金哲,姚艳飞等.梅花鹿鹿花盘对小鼠抗疲劳作用的研究[A].吉林农业大学学报,2011,1000-5684.
    [14]邱芳萍,马波,王志兵等.鹿角盘蛋白的分离纯化与活性研究[J].长春工业大学报,2007,28(3):144-147
    [15]黄金凤,王维,王莘.鹿茸及鹿花盘蛋白提取物的抑菌比较[J].吉林农业,2010,247(9):43,45.
    [16]郜玉钢,于文影,李然等.鹿角盘多糖抗病毒的研究[J].安徽农业科学,2010,38(22):11857-11858.
    [17]陆燕.鹿角冒对大鼠骨质疏松症治疗作用的研究[D].大连:大连理工大学生命学院,2010.
    [18]张爱红,梓醇对脂多糖诱导的小鼠炎症反应的调节作用[D].大连:大连理工大学生命学院,2008.
    [19]MANN DL. Inflammatory mediators in heart failure:homogeneity through heterogrnrity [J]. Lancet,1999,353(9167):1812-1813.
    [20]赵德明.兽医病理学第二版[M].北京.中国农业大学出版社,2005:133-157.
    [21]何龙.细胞因子自分泌现象及其生理和病理意义[J].上海免疫学杂志,1994,14(3):184-187.
    [22]NORMAN J. The role of cytokines in the pathogenesis of acute pancreatitis[J]. Am J Surg,1998,175:76-83.
    [23]涂亚庭,林能兴.细胞因子在皮肤病治疗中的现状和展望[J].中华医学杂志,2005,29(3):145-146.
    [24]MAKHILA R, KINGSNORTH AN. Cytokine storm in acute pancreatitis[J]. Hepatobiliary Pancreat Surg,2002,9:401-410.
    [25]汤亲青,赵怀辉,李钟声.细胞因子在胰腺炎发生发展中的作用[J].肝胆外科杂志,2009,17(5):397-399.
    [26]XU SY. The pharmacological progress of anti-inflammatory agents and drugs acting on immunity[J]. In:Lei HP, eds. Proceedings of the international sysposium on traditional medicine and modern pharmacology, Beijing,1986:163.
    [27]徐叔云,丁长海.抗炎免疫药物的临床研究进展[J].中国临床药理学杂志,1997,13(4):238.
    [28]HAWKEY CJ. COX-2 chronology[M]. Gut,2005,54:1509-14.
    [29]JONES R. Nonsteroidal anti-inflammatory drug prescribing:past, present and future[J]. Am J Med,2001,110:4S-7S.
    [30]JACK DB.One hundred years of aspirin[M]Lancet,1997,350:437-9.
    [31]ADAMS SS. The discovery of brufen[J]. Chem Br,1987,23:1193-5.
    [32]JASANI MK, DOWNIE WW, SAMUELS BM, et al. Ibu-profen in rheumatoid arthritis. Clinical study of analgesic and anti-inflammatory activity [J]. Ann Rheum Dis,1968, 27:457-62.
    [33]王新,张秀英.9种中药抗炎作用的实验观察[J].中国兽医杂志,2007,43(1):25-26.
    [34]阳洁,刘华刚.中药抗炎作用分子机制研究进展[J].广西科学,2005,12(3):208-213.
    [35]杨牧祥,方朝义,谷振勇等.咳喘宁胶囊对慢性支气管炎大鼠血清,肺组织及支气管肺泡灌洗液SOD, CAT活性及MDA含量的影响[J].中国中医基础医学杂志,2002,8(1):14-18.
    [36]罗仕华,符诗聪.中药抗炎作用机制的研究进展[J].药学服务与研究,2003,1:47-49.
    [37]张永健,苏素文,谢彦华等.雷公藤多甙的抗炎作用与NO的关系[J].中国药学杂志,2002,35(1):20-23.
    [38]牛颖,方邦江,周爽等.锦红汤对急性胆源性感染中全身性炎症反应免疫调节作用的影响[J].湖北中医杂志,2004,26(3):12-14.
    [39]王秀丽,李冬晖,高春芳.商陆皂甙甲对小鼠腹腔巨噬细胞分泌NO自由基的影响[J].实用医药杂志,2004,21(7):628.
    [40]刘金霞,邓淑华,杨贺松等.黄芩茎叶总黄酮的抗炎作用机制的研究[J].中国药理学通报,2002,18(6):713-714.
    [41]周军,方素萍,齐云等.葛根汤对佐剂性关节炎大鼠关节液炎症介质的影响[J].中国实验方剂学杂志,2001,7(3):29-31.
    [42]杨明炜,陆付耳,李呜真.金叶败毒颗粒抗炎与免疫药理作用的研究[J].中国实验方剂学杂志,2000,6(3):26-28.
    [43]翁玲玲,楼荣良.国外正在研制的甾体药物[J].中国药学杂志,1994,3(29):174-176.
    [44]黄炎.长期应用糖皮质激素的毒副作用[J].中国实用儿科杂志,2000,15(1):15-17.
    [45]刘芬,刘洁,陈霞等.氧化苦参碱的抗炎作用及其机制[J].吉林大学学报,2005,31(5):728-730.
    [46]黄利权,伍义行.火绒草的抗炎活性研究[J].中兽医学杂志,2004,117(2):10-12.
    [47]王文静,王军,饶高雄.接骨草的两种提取物对小鼠的抗炎镇痛作用[J].华西药学杂志,2011,3:247-249.
    [48]梅全喜,戴卫波,曾聪彦等.布渣叶水提物抗炎作用的实验研究[J].国际中医中药杂志,2010,32(1):16-17.
    [49]马凤余,鄢顺琴,凤良元.胃舒灵Ⅱ抗炎作用与机制研究[J].安徽中医学院学报,2003,3:41-44.
    [50]吴勇杰,常珍梅.甘草次酸钠的抗炎作用机理[J].中国药理学通报,1991,1:46-49.
    [51]王文魁,沈映君,齐云.辛夷精油的抗炎作用[J].陕西中医学报,2000,23(2):40-42.
    [52]匡荣,包文芳,赵明宏等.文冠木正丁醇提取物的抗炎作用[J].沈阳药科大学学报,2001,18(1):53-56.
    [53]张白嘉,刘亚欧,刘榴等.土茯苓及落新妇苷抗炎,镇痛,利尿作用研究[J].中药药理与临床,2004,20(1):11-12.
    [54]陈长勋,刘占文,孙峥嵘等.龙胆苦苷抗炎药理作用研究[J].中草药,003,34(9):814-816.
    [55]曾南,沈映君,刘旭光等.荆芥挥发油抗炎作用研究[J].中药药理与临床,1998,14(6):24-26.
    [56]祁红.大黄素的抗炎作用[J].中草药,1999,7:522-524.
    [57]BECKMAN JS, BECKMAN TW, CHEN TW, et al. Apparent hydroxyl radical production by peroxynitrite:implication of endothelial injury from nitric oxide and super-oxide [J]. Proc Natl Acad Sci USA,1990,87:1620-1624.
    [58]RADI R, BECKMAN JS, BUSH KM, et al. Peroxyni-trite-induced membrane lipid peroxidation:the cytotoxic potential of superoxide and nitric oxide [J]. Arch Biochem Biophys,1991,288:481-487.
    [59]RUBBO H, RADI R, TRUJILLO M, TELLERI R, et al. Freeman BA Nitric oxide regulation of superoxide and peroxynitrite-dependent lipid peroxidation [J]. J BiolChem 1994, 269:26066-26075.
    [60]DI ROSA M, WILLOUGHBY. DA Screens for anti-inflammatory drugs [J]. J Pharm Pharmacol 1971,23:297-300.
    [61]YOSHIKAWA T, KONDO M, YAGI K. Lipid peroxide and disease [M]. Igaku-shoin, Tokyo, 1981:210.
    [62]SYMONS AM, DOWLING EJ, PARKE DV. Lipid peroxidation free radicals and experimental inflammation [J]. Basic Life Sci,1988,49:987-990.
    [63]CLAXSON A, MORRIS C, BLACK D, et al. The anti-inflammatory effects of D-myoinositol-1,2,6-triphosphate (PP56) on animal models of inflammation [J]. Agents Actions,1990,29:68-70.
    [64]GRIFFITHS HR, DOWLING EJ, SAHINOGLU T, et al. The selective protection afforded by ebselen against lipid peroxidation in a ROS-dependent model of inflammation [J]. Agents Actions,1992,36:107-111.
    [65]李焕德,彭文兴.抑郁症药物治疗新进展[J].国外医学精神病学分册,1997,24(3):129-133.
    [66]毛榕榕,田孟,徐林.抑郁的大脑:抑郁症的神经生物学研究和抗抑郁药物新药的研发[J].自然杂志,2009,31(3):145-152.
    [67]罗和春.漫谈抑郁状态[J].心理与健康,1994,(5):4-5.
    [68]HANKIN BL, ABRAMSON LY. Development of gender differences in depression:an elaborated cognitive vulnerability-transactional stress theory [J].Psychol Bull, 2001,127(6):773.
    [69]易正辉,王贻儒,王祖承.抑郁症神经生化和神经电生理学研究进展[J].中国新药与临床杂志,2005,24(9):67-678.
    [70]BERRIDGE CW, WATERHOUSE BD. The loeus coeruleus-noradrenergic systerm:modulation of behavioral state and statedependent cognitive processes [J]. Brain Res Rev.2003, 42:33-84.
    [71]蔡伯艳,谢海芳,愈雯雯.抑郁症与神经递质关系的研究进展[J].辽宁中医药大学学报,2007,9(5):51-52.
    [72]THOMSEN AF, KVIST TK, ANDERSON PK, et al. Inereased risk of affective disorder following hospitalization with hyperthyroidism aregister-Based study [J]. Eur J Endocrinol,2005,152(4):535-543.
    [73]GIAN Ml, VINCENZO M, FILIPPO D. Inereased sensitivity to antidepressants of D3 dopamine receptor-deficient mice in the forced swim test(FST)[J]. European Neuropsychopharmacology,2008,18(4):271-277.
    [74]方允中,李文杰.自由基与酶0基础理论及其在生物学和医学中的应用[M].北京,科学出版社,1994.
    [75]WEJTASZEK P. Oxidative burst:an early plant response to pathogen infection [J]. Biochem J,1997, (322):681-692.
    [76]UYSAL M, SECKIN S, KOCAK-TOKER N et al. Increased hepatic lipid peroxidation in aged mice [J]. Mech Ageing Dev,1989,48:85.
    [77]王文杰.超氧自由基和超氧化物歧化酶[J].生理科学进展,1985,16(3):196.
    [78]MANN T, KEILIN D. Haemocuprein and hepatocuprein, copper-protein compounds of blood and lier in mammals[J]. Proc R Soc B,1938,126:303-315.
    [79]MC CORD J M, JIWIN FRIDOVICH. Superoxide dismutase, An enzymatic function for erythrocuprein (hemocuprein) [J].J Biol Chem,1959,244:6049-6055.
    [80]马森.谷胱甘肽过氧化物酶和谷胱甘肽转硫酶研究进展[J].动物医学进展,2008,29(10):53-56.
    [81]楼善贤.溶菌酶的研究进展[J].中国肿瘤临床,1994,21(9):709-711.
    [82]王佃亮.重组人溶菌酶研究进展[J].中国生物工程杂志,2003,23(9):59-63.
    [83]董吉祥,张学光,刘志达等.慢性肾炎患者血清IL-2,IL-6及其受体水平变化研究[J].苏州大学学报,2002,4:360-362.
    [84]BEUTLER B. Innate immunity:an overview [J]. Mol Immunol,2004,40:845-859.
    [85]赵桂芝,聂淑琴,杨庆等.清脂胶囊对内毒素诱导的炎症介质水平的影响及时效关系[J].中国实验方剂学杂志,2006,11(12):39-42.
    [86]王旭光,陈根殷,陈妙萍.木犀草素对LPS诱导的RAW264.7细胞COX02及mPGES-1表达的影响[J].中药材,2007,30(10):1263-1266.
    [87]黄峰,王立为.中药抗炎免疫药理研究进展[N].新乡医学院学报,2002,19(4):340-343.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700