摘要
肥胖是一种以身体脂肪含量过多为重要特征的,多病因的,能够并发多种疾病的慢性病,与艾滋病、吸毒、酗酒并列为世界性四大医学社会问题,并可导致脂质代谢异常,胰岛素抵抗和高血脂、高血压、睡眠呼吸暂停症、脂肪肝、胆结石、脑卒,骨关节病、癌症等疾病,随着肥胖人口的日益增多,肥胖已经成为我国及广大发达及发展中国家面临的一个严重的公共健康问题。目前,治疗肥胖的西药品种繁多,疗效也较明显,但因其副作用强,不良反应多,大多不适于长期使用或已禁止使用。
近几年来,中医学对肥胖病进行了广泛而深入的研究,也取得了较大的进展。又因为中医治疗肥胖症的副作用小、反弹率低,越来越受到国内外学者的重视。从中药提取物的国际市场现状来看,随着中药提取物在“替代疗法”方面的广泛应用,人们越来越意识到,许多传统的草药比临床使用的西药疗效更好,更具有安全性,国际市场需求不断增大。而我国植物提取物的年出口额在中药出口商品中的构成比呈明显的上升趋势,其国际市场空间也在不断扩大。
荷叶系睡莲科植物莲的干燥叶,具有清热解暑,升发清阳,凉血止血之功效,其分布范围广泛,产量巨大。其主产地于中南地区及江浙一带,尤其是湖北省具有悠久的种植历史和丰富的栽培资源。荷叶含有槲皮素、荷叶碱等多种化学成分,被广泛应用于具有减肥降脂功能及治疗单纯性肥胖症的中药单复方制剂中,具有良好的临床效果。
由于荷叶潜在的临床使用价值,加之鄂产荷叶来源广泛,质量良莠不齐,尚缺乏全面、统一的品质评价手段,给临床应用造成诸多不便。本文采用HPLC法测定槲皮素和荷叶碱的含量,同时采用亚硝酸钠-硝酸铝-氢氧化钠比色法和酸性染料比色法测定其总黄酮和总生物碱的含量,结果显示汉川、潜江、武汉汉阳荷叶的指标成分含量较高,九月为荷叶采收的最佳时期。通过多指标综合评价荷叶药材质量,弥补了《中国药典》中关于使用荷叶碱单一质控指标的局限性。
在明确鄂产荷叶各有效成分含量的基础上,结合大孔吸附树脂技术提取分离其黄酮部位,以槲皮素为指标,采用正交试验优化黄酮提取物的工艺条件,选择适宜的树脂类型,并考察供试品溶液pH值、吸附流速、吸附液浓度、径高比、泄露曲线、清洗液pH值、洗脱液浓度、洗脱液用量等因素对黄酮部位分离的影响,结果表明:选择HPD100型树脂对荷叶黄酮组分进行吸附与解吸性能较好,采用吸附液浓度为0.2g·mL~(-1)、流速为4BV·h~(-1)的、吸附液pH为1～3、径高比为1:9、吸附液体积为柱床体积的11倍参数组合应用效果良好,清洗液pH小于5、2BV 50%乙醇洗脱效果较好且经济省时。
按上述工艺流程分离制备荷叶黄酮提取物,通过建立大鼠营养性肥胖模型,考察荷叶黄酮提取物对于营养性肥胖大鼠及血脂指标的影响,结果表明,荷叶黄酮部位可明显抑制大鼠体重的增加,降低其Lee's指数及脂肪重量,其中高剂量组能显著降低TC、TG水平,升高HDL-C/LDL-C,与模型组比较具有显著性差异。因此荷叶黄酮具有较好的减肥效果,而对动物体质耐力无明显影响。
总之,通过鄂产荷叶中黄酮、生物碱化合物成分的分析以及大孔吸附树脂分离纯化黄酮工艺流程的优化,为评价荷叶减肥降脂相关活性提供了必要前提,从而为进一步阐明其分子机制提供了科学依据,并为今后建立化学指标和生物学指标相结合的质量评价体系奠定了基础,在确定荷叶品系品质的基础上对其进行规模化种植,开发新型具减肥降脂功效的保健食品,有利于加快鄂产荷叶的经济开发利用;同时增加中部地区莲农的经济收入,促进当地经济社会和谐发展。
Overweight and obesity are defined as abnormal or excessive fat accumulation that presents a risk to health and are major risk factors for a number of chronic diseases.It is one of four biggest social and medical problems following AIDS,drug taking and excessive drinking.It can interferes with lipid metabolism,induces insulin resistance,hyperlipemia,hypertension,sleep apnea,fatty liver,gallstone,osteoarthrosis,cancer,etc.Nowadays,obesity patients are more and more,it has already been a significant public health problem that our country and many developed and developing countries are facing.At present,western medicines are numerous and effective,but they are not suitable for long-term use or has been forbided because of obvious side effect and untoward reaction.
In recent years,therapy of obesity was studied extensively and profoundly in traditional Chinese medicine and has made important progress.Because of less side effects and low recurring rate,scholars all over the world have paied more and more attentions on obesity treatment with traditional Chinese medicine.Besids,because of vicarious treatment has been used widely in international market,more and more people notice that lots of traditional Chinese medicines are safer and more effective than western medicine.Its international market demand are widely increasing.In recent years,ratio of extract in export products of traditional Chinese medicine is ascending in our country.Its international market potential are extensive.
Folium Nelurnbinis is the dried leaf of Nelumbo nucifera Gaertn.It has the effect of clearing away summer-heat and heat evil,ascending the lucid yang and cooling blood to stop bleeding.It grows in middle and south of China and the region of Zhejiang and jiangsu.Especially in Hubei province,it has a long history of cultivation and has abundance resources.Folium Nelumbinis contains kinds of chemical ingredients and has been added to single and compound recipes treating obesity clinically.
Because of the potential clinical value of Foliurn Nelumbinis and the extensive resource,its quality is difficult to control.Being lack of unified estimated standard causes much inconvenience in clinical application.The thesis aims to establish methods with HPLC,Sodium Nitrite-aluminum nitrate-sodium hydroxide colorimetry and acid-dye colorimetry to determine the content of quercetin, nuciferine,total flavonoids and total alkaloids in Folium Nelumbinis respectively.The results show that the contents of index ingredients in areas of Hanchuan,Qianjiang and Hanyang were relatively higher than that from other habitats,and the month of September is the best time to collect.
On the basis of confirming the index ingredients,the total flavonoids were extracted with macroporous adsorptive resins from Folium Nelumbinis.The process was optimized by employing orthogonal test design with the content of quercetin as the index.The resin HPD 100 had much better adsorption capacity and desorption ratio by using 2 BV 50%aqueous ethanol to extract the flavonoids from Folium Nelumbinis than other resins.The results showed that HPD100 possessed the highest absorption capacity under the concentration of sample 0.2g·mL~(-1),the condition of flowing rate of 4BV·h~(-1).Macroporous adsorptive resin HPD100 may be value of isolating and purifying the flavonoids from FoIium Nelumbinis.
The flavonoids extract was prepared according to the technological process above,and the effect on the trophism obesity rat and blood-fat index was investigated.Tile result showed that the rats' weight,Lee' indexes,and the weight of rats' fat was obviously descended.It also can reduced TC and TG,yet improve the HDL-C/LDL-C in blood serum.Thus it can be seen that the flavonoids extract of Folium Nelumbinis have significant effect on the reduction of fat and blood lipids.
Generally speaking,through the analysis of flavones and alkaloids in Hubei province and optimized method of extraction of flavonoids with macroporous adsorptive resins,it supplied the essential precondition to evaluate the activity of anti-obesity and degrading the blood-fat with Folium Nelumbinis.Simultaneously,it is possible to elucidate its molecule mechanism with concerned experimental data.It established basis to set up the system of quality evaluation combining the chemistry and biology index.It is in favor of the promote of Folium Nelumbinis economy in Hubei province.It could develop the novel nutritious food and drinks,which will increase the income of peasants who plant Folium Nelumbinis.
引文
[1]张颖,张晓丹.肥胖的发病机制及其中药治疗概况[J].中医药信息.哈尔滨:2007,24(5):13-15.
[2]陈建双,赵蕾,袁亚非.肥胖症研究进展[J].承德医学院报.2007,24(2):191-193.
[3]文发魁,苏东峰.肥胖与血糖、血脂、血压的相关性研究[J].中国社区医师.京:2007,(14):97.
[4]WANG Lin-ping.The relationship of obesity and myocardial diseas[J].Molecular Cardiology of China.Beijing:2007,7(5):310-313.
[5]XIA Yu,HUANG Ying.Advance on studies of obesity and asthma[J].International Journal of Respiration.Shijiazhuang:2007,27(11):866-869.
[6]XIE Ai-xia,WU Sheng-li,LI Nong.Observation of the relationship of obesity with diabetes and hypertension[J].Chinese Journal of Diabetes.Beijing:2007,15(9);556-557.
[7]陈东方,陶广凤.肥胖疾病谱[J].医药世界.北京:2007,(8):43-45.
[8]杜娟,马俐儒,罗萍,等.肥胖患者肾脏病理改变及临床特点分析[J].中国实验诊断学.长春:2007,11(8):1109-1111.
[9]钟华荣,吕安康,沈卫峰.从分子机制探讨肥胖致动脉粥样硬化[J].国际心血管病杂志.上海:2007,34(5):327-330.
[10]Zhong ZH,Cheng CY,He YH,etc.Relationship of overweight and obesity to fatty liver:A case-control study[J].Journal of Clinical Rehabilitative Tissue Engineering Research.Shenyang:2007,11(39):7773-7776.
[11]XIE Yan-hong,HAN Cun-zhi.Recent progress in the relationship between obesity and Cancer[J].International Journal of Endocrinology and Metabolism.Tianjin:2007,27(1):59-61.
[12]饶向荣,张改华.肥胖与肾脏损害[J].中国中西医结合肾病杂志.太原:2006,7(10):616-618.
[13]张颖,张晓丹.肥胖的发病机制及其中药治疗概况[J].中医药信息.哈尔滨:2007,24(5):13-15.
[14]马培奇.抗肥胖药奥利司他和西布曲明概述[J].中国制药信息.北京:2000,16(10):28-29.
[15]张京芳,吕宪祥,刘志民.氛氟拉明的一般药理及毒理作用[J].中国药物依赖性杂志.北京:2002,41(4):316-318.
[16]曹晨昱.肥胖症的药物治疗及其进展[J].中国药师.武汉:2007,10(11):1077- 1079.
[17]ZHAI Yun-qi,WU Ying-liang.Pharmacological review of sibutramine and its optical isomers[J].Journal of Shenyang Pharmaceutical University.Shenyang:2003,20(3):222-225.
[18]朱玛华,应翔宇.西布曲明引起健忘症的信号分析[J].药物警戒.北京:2005,2(1):51-52.
[19]曾涛.常见减肥药物的不良反应[J].中国药师.武汉:2007,10(6):599-601.
[20]孙忠实,朱珠.药物不良反映报告5例[J].中国医药导刊.北京:2001,3(2):149-150.
[21]陈应兰,王国村.新型减肥药奥利司他的药理及临床应用[J].中国药房.重庆:2001,12(3):184-186.
[22]B.Beermann,H.Melander,J.sawe,etc.Inccorrect use and limited weight reduction of orlistat(Xenical)in clinical practice[J].European Journal of Clinical Pharmacology.Heidelberg:2001,57:309-311.
[23]MARK FOX.MD,BERNADETTE STUTZ,DIETER MENNE PhD,etc.The Effects of Loperamide Continence Problems and Anorectal Function in Obese Subjects Taking Orlistat[J].Digestive Diseases and Sciences.Netherlands:2005,50(9):1576-1583.
[24]金光亮.肥胖症的中医治疗[J].日本医学介绍.沈阳:1994,15(2):95-96.
[25]张志军,陈贵廷,黄欣.中医药降脂减肥再哟用的国内外研究进展[J].中国中医药信息杂志.北京:1995,2(1):14-17.
[26]ZHAO Jin-jun,CHEN Yu-yao,TONG Li.Treatment by Chinese herbs on adiposity [J].Modem Rehabilitalion.Shenyang:2001,5(9):18-20.
[27]孙丰雷,郎江明.中医药防治肥胖症进展[J].国际医药卫生导报.广州:2002,(10):2-4.
[28]郭澄,魏道智,陈海华.中医药减肥疗法的研究概况[J].中药材.广州:2002,25(7):534-537.
[29]孙升云,王风珍.中医药防治单纯性肥胖的临床研究进展[J].时珍国医国药.北京:2006,17(3):331-332.
[30]邢宁,何生华.单纯性肥胖症的中医治疗[J].时珍国医国药.北京:2006,17(10):2046-2047.
[31]徐为公,徐鸱.中药提取物的国际化策略[J].中药研究与信息.北京:2005,7(10):34-36.
[32]柯卫东,李峰,刘玉平等.我国莲资源及育种研究综述[J].长江蔬菜.武汉:2003, (4):5-9.
[33]党毅,肖颖,主编.中药保健食品研制与开发[M].人民卫生出版社.北京:2002,19.
[34]Ono Y,Hattori E,Fukaya Y,etc.Anti-obesity effect of Nelumbo nucifera leaves extract in mice and rats[J].J Ethnopharmacol,2006,106(2):238-244.
[35]田娜,刘仲华,黄建安.莲的化学成分及其药理作用研究进展[J].中国药学杂志.北京:2005,40(16):1212-1214.
[36]刘密新,吴筑平,杨成对等.荷叶精油与生物碱的分析研究[J].清华大学学报(自然科学版).北京:1997,37(6):35-37.
[37]刘淑萍,樊淑彦,候海妮等.荷叶化学成分及药理作用研究进展[J].河北医科大学学报.石家庄:2004,25(4):254-256.
[38]Elegami AA,Bates C,Gray AI,et al.Two very unusual macrocyclic flavonoids from the water lily Nymphaea lotus.[J].Phytochemistry.2003,63(6):727-731.
[39]杜力军,孙虹,李敏等.荷叶大豆及其合剂调脂活性部位的研究[J].中草药.天津:2000,31(7):526-528.
[40]陶波,帅景贤,吴凤莲.荷叶水煎剂对高脂血症大鼠血脂及血液流变学的影响[J].中医药学报.重庆:2000,(6):55-56.
[41]涂长春,李晓宇,杨军平等.荷叶生物总碱对肥胖高脂血症大鼠减肥作用的实验研究[J].江西中医学院学报.南昌:2001,13(3):120-121.
[42]YANG Xin-yue,ZHOU Yin-ping,DAI Li-guang,etc.Antilipemic Effect of Extracts of Ampelopis Grossedentata and Lotus Leaf(EAGLL)on Rats with Hypercholesterole-mia[J].CHINESE JOURNAL OF COMPARATIVE MEDICINE.Beijing:2005,15(5):281-284.
[43]ZHAO Jun,GAO Lan,QI Zhao-peng.Extraction of Folium Nelumbinis Total Alkaloid and Its Salt and Comparison in Lowering Blood Lipid between them[J].Tianjin Journal of Traditional Chinese Medicine.Tianjin:2005,22(2):161-162.
[44]柏田良树等.抗HIV天然物的研究(5):荷叶的抗HIV成分[J].国外医学中医中药分册.北京:1997,19(6):45.
[45]HUANG Agen,SHI Hong-fei,WEI Hong,etc.A Comparative Study on Blood Lipid Regulatory Function Between Flavonoids and Alkaloids from Lotus Leaf [J].Culinary Science Journal of Yangzhou University.Yangzhou:2006,23(3):23-25.
[46]CHEN Gang,SU Wei.STUDY ON THE ANTIHYPERLIPIDEMIA OF THE EXTRACT OF LOTUS LEAF BY SUPERCRITICAL CARBON DIOXIDE[J].Food Research and Development.Tianjin:2006.27(11):62-64.
[47]龚康敏,魏克民,厉兰娜等.荷叶饮预防大鼠高脂血症及相关指标实验观察[J].浙江中西医结合杂志.杭州:2001,11(3):148-149.
[48]唐春红,幸宏伟,石轶松等.黄酮提取液对大鼠高脂血症及血液流变学参数的影响[J].第三军医大学学报.重庆:2004,26(7):616--17.
[49]GUO Huii-min,CHEN Mei-zhen,YU Jie,etc.Experimental Study on the Antiobesity Action of Fat-reducing Tea Compounded from Edible Medical Plants[J].Journal of Shantou University(Natural Science Edition).Shantou:18(2):26-29,34.
[50]陈海光,余以刚,曾庆孝.荷叶保健茶的研制及其功效研究[J].食品工业科技.广州:2002,23(1):53-54.
[51]陈奇,主编.中药药效研究思路与方法[M].人民卫生出版社.北京:2004,629.
[52]纪丽莲.荷叶中抑菌成分的提取及其抑菌活性的研究[J].食品科学.北京:1999,(8):64-66.
[53]Tang Yufang,Zhang Miaolin,Liu Zhongyi,etc.Study of Abstraction and Antimicrobial Activities..of Lotus Leaf″S Alkaloid[J].Guangzhou Food Science and Technology.Guangzhou:2005,20(2):51-52.
[54]CHEN Jian-fen,HE Wei-hua,QiAN Fu-gang.Qualitative and quantitative analysis of bacteriostatic constituents of lotus leaf extract for mouth cavity[J].China Surfactant Detergent & Cosmetics.Taiyuan:2003,33(1):49-52.
[55]唐裕芳,张妙玲.荷叶超临界CO_2萃取物抑茵效果稳定性研究[J].食品科技.广州:2004,(12):53-55.
[56]蒋益虹.荷叶抑菌有效成分的提取优化工艺研究[J].中国中药杂志.北京:2005,30(13):1030-1031.
[57]Zhang Yunbin,Li Caixia.Studies on Antoxidative and Antimicrobial Activities of Ethanolic Extract from Lotus Leaf[J].FOOD AND FERMENTATION INDUSTRIES.Beijing:2005,31(10):21-24.
[58]蔡为荣.微波提取荷叶黄酮及其清除羟基自由基的研究[J].食品科学.北京:2004,25(9):113-115.
[59]Wu MJ,Wang L,Weng CY,et al.Antioxidant activity of methanol extract of the lotus leaf(Nelumbo nucifera Gertn.)[J].Am J Chin Med.2003,31(5):687-698.
[60]Rai S,Wahile A,Mukherjee K,et al.Antioxidant activity of Nelumbo nucifera (sacred lotus)seeds[J].J Ethnopharmacol.2006,104(3):322-327.
[61]Sohn DH,Kim YC,Oh SH,et al.Hepatoprotective and free radical scavenging effects of Nelumbo nucifera[J].Phytomedicine.2003,10(2-3):165-169.
[62]肖华山,黄代青,傅文庆等.荷叶对体外氧自由基的清除作用及其对果蝇寿命的 影响[J].中国老年学杂志.长春:1996,16(6):373-375.
[63]Chen Haiguang,Zeng Qingxiao.Study on the Extracting Technology of Functional Components from Lotus Leaf and the Effect on Scavenging Oxygen Free Radicals[J].Food and Fermentation Industrie.Beijing:2001,27(10):34-38.
[64]余以刚,陈海光,曾庆孝.荷叶水提物清除自由基的ESR研究[J].中草药.天津:2001,32(8):693-695.
[65]纪丽莲,范怡梅.荷叶提取物抗氧化活性的研究[J].淮阴工学院学报.淮安:2001,10(1):12-14.
[66]柏田良树等.抗HIV天然物的研究(5):荷叶的抗HIV成分[J].国外医学中医中药分册.北京:1997,19(6):45.
[67]Kashiwada Y,Aoshima A,Ikeshiro Y,et al.Anti-HIV benzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera,and structure-activity correlations with related alkaloids[J].Bioorg Med Chem.2005 Jan 17;13(2):443-448.
[68]苗明三.荷橘茶药效学研究[J].中医研究.郑州:1997,10(3):22-24.
[69]陈德兴,袁秀荣,毛平等.荷叶在降脂减肥食疗制剂中的应用[J].药膳食疗.长沙:2002,(10):5-9.
[70]GUAN Zhang-shun,WU Jun,YU Ze-lan,etc.Effects of lotus leaf capsule on human's blood lipids disorde[J].CHINESE JOURNAL OF CARDIOVASCULAR REHABILITATION MEDICINE.Fuzhou:2003,12(4):294-297.
[71]李慧芬.荷丹片治疗高脂血症80例疗效观察[J].中国社区医师.北京:2006,8(2):34.
[72]张建伟,顾勇主.自拟祛脂汤治疗脂肪肝90例[J].陕西中医.西安:2002,23(1):10-11.
[73]连粤湘,李梅芳,岑玉文等.荷叶汤联合肝病治疗仪治疗脂肪肝36例疗效观察[J].新中医.广州:2005,37(5):36-37.
[74]雷载权,张延模主编.中华临床中药学[M].人民卫生出版社.北京:1998,413-433.
[75]史晓岚.地榆荷叶在产后泄泻中的应用[J].中华腹部疾病杂志.泰安:2003,3(3):209.
[76]赵正保,韩锋,曲婷丽等.高效液相色谱法测定消渴降糖胶囊中槲皮素的含量[J].中国药物与临床.太原:2004,4(12):919-921.
[77]刘东平,周亚球,王先荣.罗布麻叶总黄酮的测定方法探讨[J].时珍国医国药.北京:2007,18(3):614-616.
[78]卫星星.苦荞籽壳中黄酮类化合物的定性定量研究[J].数理医药学杂志.武汉:2007,20(4):537-538.
[79]刘文英主编.药物分析[M].人民卫生出版社.北京:2003,194.
[80]Grohmann K,Manthey JA,Cameron RG,et al.Purification of citrus peel juice and molasses.J Agric Food Chem,1999,47(12):4859-67.
[81]盛萍,帕丽达·阿不力孜,刘波,等.大孔树脂吸附法富集野菊花总黄酮的工艺研究[J].中草药.北京:2006,37(8):1170-1173.
[82]傅冬和,刘爱玲,邓克尼,等.DM-130树脂对甘草酸的吸附性能及提纯应用研究[J].天然产物研究与开发.成都:2002,14(1):60-64.
[83]杨玺,主编.专家解答肥胖病[M].上海科学技术文献出版社.上海:2004,3
[84]李春生,主编.现代肥胖病学[M].科学技术文献出版社.北京:2004,4
[85]徐叔云,卞如濂,陈修主编.药理实验方法学(第三版)[M].人民卫生出版社.北京:2001,1189.
[86]Xie Wei,Yang yong-liang,Liang Li.Effent of gingko leaf extract on delaying sports fatigue[J].CHINESE JOURNAL OF CLINICAL REHABILITATION.Beijing:2005,9(36):162-165.
