半枝莲抗癌成分的研究
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摘要
半枝莲(Herba Scutellariae Barbatae)为唇形科(Labiatae)黄芩属植物半枝莲Scutellaria barbara D.Don的干燥全草。分布于华东、华南、西南及河北、河南、陕西南部、湖北、湖南等地。半枝莲味辛、苦、性寒。归肺、肝、肾经。是中医处方和成药中的常用中药。具有清热解毒、化瘀利尿的功效,用于疔疮肿毒、咽喉肿痛、毒蛇咬伤、跌扑肿痛及水肿、黄疸等。
癌症又称恶性肿瘤,是目前危害人类健康的主要疾病之一,近年来,从中药提取的抗肿瘤有效成分日益得到人们的重视。半枝莲是近几年国内外抗肿瘤药的研究热点之一。现代药理研究表明,半枝莲水提物和醇提物抗肿瘤的作用机制主要包括:直接抑制肿瘤生长与增殖作用、诱导肿瘤细胞凋亡作用、抗氧化抗自由基作用等。但已有文献中有关半枝莲及其化学成分的抑制血管生成作用方面的研究很少,有少量体外活性试验的报道,尚无人用活体动物开展半枝莲及其提取物抑制血管生成活性方面的研究。本文利用斑马鱼抑制血管生成模型对半枝莲化学成分进行了逐级筛选,迄今为止未见报道。
本文对半枝莲75%乙醇提取物进行了系统研究,首先采用溶剂萃取法对其行了初步分离,得到不同极性的三部分,分别是石油醚层提取物、乙酸乙酯层提取物、正丁醇层提取物。利用斑马鱼抑制血管生成模型对三种提取物进行了活性测定,确定了乙酸乙酯层提取物有抑制血管生成活性。然后采用硅胶柱色谱,凝胶柱色谱,重结晶等手段,对乙酸乙酯层提取物进行了进一步的分离,得到了13种化合物,并通过理化性质和光谱数据分析确定其化学结构,这些化合物分别为:β-谷甾醇、汉黄芩素、4’-羟基汉黄芩素、芹菜素、黄芩素、柚皮素、木犀草素、异鼠李素、槲皮素、β-胡萝卜苷、野黄芩苷、黄芩苷、木犀草苷。最后利用斑马鱼模型对这13种化合物的抑制血管生成活性分别进行了测定,确定异鼠李素和槲皮素具有抑制血管生成活性,并且随着样品浓度增大抑制血管生成活性随之增强。在100μg/mL时,抑制率分别达到76.79%和61.24%。
Herba scutellariae barbatae is the dried whole plant of Scutellaria barbata D. Don which belongs to Scutellaria Labiatae, distributed in the east, south, southwest China and Hebei, Henan, southern Shaanxi, Hubei, Hunan and other places. Herba scutellariae barbatae is spicy, bitter in flavor and cold in property, and it is attributive to the lung meridian, live meridian and kidney meridian. It’s a commonly used traditional Chinese medicinal material in prescription to clear away heat and toxic materials, resolve stasis and diuretic, and used for swollen boils, sore throat, snake bites, flutter swelling, edema, and jaundice.
Cancer, also known as malignant tumors, is one of the major diseases harming human health. The anti-tumor active ingredients extracted from traditional Chinese medicinal materials have been paid more and more attentions in recent years. Herba scutellaria barbatae has been one of the main focus in researches on anticancer drugs at home and abroad in recent years. Modern pharmacological studies have shown that the extracts from herba scutellaria barbatae by water and ethanol have significant anti-tumor effects in different mechanisms, including inhibiting tumor growth and proliferation directly, inducing tumor cell apoptosis, antioxidation and clearing free radicals. There are few published papers about antiangiogenic activity of herba scutellariae barbatae and its chemical compositions with a few in vitro tests and no papers about studies on antiangiogenesis of it and its extracts on basis of live animals. Screening active compounds from herba scutellariae barbatae by using zebrafish antiangiogenesis model in this paper hasn’t been reported currently.
In this paper, systematic studies on 75% ethanol extracts has been finished. Firstly, three parts with different polarities were got by solvent extraction, which are petroleum ether extract, ethyl acetate extract and butanol extract. After screening with zebrafish model, the ethyl acetate extract was determined to have antiangiogenic activity. Secondly, 13 compounds were got from ethyl acetate extract by silica gel column chromatography, gel column chromatography, recrystallization and etc. These compounds were determined to beβ-sitosterol, wogonin, 4'-hydroxy wogonin, apigenin, baicalein, naringenin, luteolin, isorhamnetin, quercetin,β-daucosterol, wild baicalin, baicalin and luteolin by means of NMR and physical and chemical properties analysis. Lastly, we used the zebrafish model to envaluate the antiangiogenic activity of these 13 compounds, and the results show that quercetin and isorhamnetin have significant antiangiogenic activities with inhibitory rate of 76.79% and 61.24% at 100μg/mL, respectively.
癌症又称恶性肿瘤,是目前危害人类健康的主要疾病之一,近年来,从中药提取的抗肿瘤有效成分日益得到人们的重视。半枝莲是近几年国内外抗肿瘤药的研究热点之一。现代药理研究表明,半枝莲水提物和醇提物抗肿瘤的作用机制主要包括:直接抑制肿瘤生长与增殖作用、诱导肿瘤细胞凋亡作用、抗氧化抗自由基作用等。但已有文献中有关半枝莲及其化学成分的抑制血管生成作用方面的研究很少,有少量体外活性试验的报道,尚无人用活体动物开展半枝莲及其提取物抑制血管生成活性方面的研究。本文利用斑马鱼抑制血管生成模型对半枝莲化学成分进行了逐级筛选,迄今为止未见报道。
本文对半枝莲75%乙醇提取物进行了系统研究,首先采用溶剂萃取法对其行了初步分离,得到不同极性的三部分,分别是石油醚层提取物、乙酸乙酯层提取物、正丁醇层提取物。利用斑马鱼抑制血管生成模型对三种提取物进行了活性测定,确定了乙酸乙酯层提取物有抑制血管生成活性。然后采用硅胶柱色谱,凝胶柱色谱,重结晶等手段,对乙酸乙酯层提取物进行了进一步的分离,得到了13种化合物,并通过理化性质和光谱数据分析确定其化学结构,这些化合物分别为:β-谷甾醇、汉黄芩素、4’-羟基汉黄芩素、芹菜素、黄芩素、柚皮素、木犀草素、异鼠李素、槲皮素、β-胡萝卜苷、野黄芩苷、黄芩苷、木犀草苷。最后利用斑马鱼模型对这13种化合物的抑制血管生成活性分别进行了测定,确定异鼠李素和槲皮素具有抑制血管生成活性,并且随着样品浓度增大抑制血管生成活性随之增强。在100μg/mL时,抑制率分别达到76.79%和61.24%。
Herba scutellariae barbatae is the dried whole plant of Scutellaria barbata D. Don which belongs to Scutellaria Labiatae, distributed in the east, south, southwest China and Hebei, Henan, southern Shaanxi, Hubei, Hunan and other places. Herba scutellariae barbatae is spicy, bitter in flavor and cold in property, and it is attributive to the lung meridian, live meridian and kidney meridian. It’s a commonly used traditional Chinese medicinal material in prescription to clear away heat and toxic materials, resolve stasis and diuretic, and used for swollen boils, sore throat, snake bites, flutter swelling, edema, and jaundice.
Cancer, also known as malignant tumors, is one of the major diseases harming human health. The anti-tumor active ingredients extracted from traditional Chinese medicinal materials have been paid more and more attentions in recent years. Herba scutellaria barbatae has been one of the main focus in researches on anticancer drugs at home and abroad in recent years. Modern pharmacological studies have shown that the extracts from herba scutellaria barbatae by water and ethanol have significant anti-tumor effects in different mechanisms, including inhibiting tumor growth and proliferation directly, inducing tumor cell apoptosis, antioxidation and clearing free radicals. There are few published papers about antiangiogenic activity of herba scutellariae barbatae and its chemical compositions with a few in vitro tests and no papers about studies on antiangiogenesis of it and its extracts on basis of live animals. Screening active compounds from herba scutellariae barbatae by using zebrafish antiangiogenesis model in this paper hasn’t been reported currently.
In this paper, systematic studies on 75% ethanol extracts has been finished. Firstly, three parts with different polarities were got by solvent extraction, which are petroleum ether extract, ethyl acetate extract and butanol extract. After screening with zebrafish model, the ethyl acetate extract was determined to have antiangiogenic activity. Secondly, 13 compounds were got from ethyl acetate extract by silica gel column chromatography, gel column chromatography, recrystallization and etc. These compounds were determined to beβ-sitosterol, wogonin, 4'-hydroxy wogonin, apigenin, baicalein, naringenin, luteolin, isorhamnetin, quercetin,β-daucosterol, wild baicalin, baicalin and luteolin by means of NMR and physical and chemical properties analysis. Lastly, we used the zebrafish model to envaluate the antiangiogenic activity of these 13 compounds, and the results show that quercetin and isorhamnetin have significant antiangiogenic activities with inhibitory rate of 76.79% and 61.24% at 100μg/mL, respectively.
引文
[1]韩玉英,丰平,文朝阳.抗癌中草药研究进展[J].北京中医,2003,22(2):45-48.
[2]国家药典委员会.中华人民共和国药典(一部) [M]北京:化学工业出版社, 2005, 77.
[3]蒋小刚.半枝莲的化学成分和药理作用[J].中国野生植物资源,2004,23(1):4-6.
[4] Lin Y-1, Chou C-J. Studies on the constituents of acrial parts of Sutslaria rivularis Wall[J]. Kou Li Chung-Kuo YaoYenChiu SoYen Chiu Pao Kao, 1984, (7), 141-143.
[5] Chou Cheng jen, Liu Kao Chu, Yang Tsang Hsiung. The isolation and characterization of scutevulin, and some other constituents from Scutellaria rivularis wall[D].Guoli Zhongguo Yiyao Yangjiuso Yanjiu Baogao, 1981 :91-108.
[6]向仁德,郑今,姚志成.半枝莲化学成分的研究[J].中草药,1982,13(8),9-12.
[7] Sonoda M, Nishiyama T, Matsukwa Y. Cytotoxic aetivities of flavonoids From two Seutellaria plants in Chinese medieine [J]. Ethnopharmeol, 2004, 91(1): 65-68
[8] Tomimori T. Chem Pharm Bull[D]. 1990, 38 (12), 3488.
[9]杨顺利,方岩雄.中药半枝莲抗肿瘤成分的分离研究[D].广州工业大学, 2002: 26-38.
[10]许益民,郭立伟,陈建伟.半枝莲多糖的分离纯化及其理化性质[J].天然产物研究与开发, 1992, 4(l): 1-5.
[11]孟研发,李志孝,张立等.半枝莲多糖SPS4组分的纯化性质及分析[J].生物化学杂志, 1993, 9(2): 224-228.
[12] Wang ZQ, Xu FM, Yan XZ, et al. Scutebarbatine A, a new neoelerodane type diterpeonid alkaloid from Seutellaria barbata[J]. Chin.Chem.Lett, 1996, 7(4): 333-334.
[13] Sheng JD, Jia YT, KeL, et al. Neo-elerodane type diterpenoides from Scutellaria barbata with Cytotoxic activities [J]. Phytoehemistry, 2006, 67: 1326-1330.
[14] Dai SJ, Chen M, Liu K, et al. Four New neo-Clerodane Diterpenoid Alkaloids from Scutellaria barbata with Cytotoxiec Aetivities [J]. Chem Pharm Bull, 2006, 54(6): 869-872.
[15] Ducki S, Hadfield JA, Lawrence NJ, et al.Isolation of E-1(4′ydroxyphenyl)-but-1-en-3-one from Scutellaria barbata[J]. Planta Med, 1996, 62(2): 185-186.
[16]席荣英,李聪辉,王翠红.半枝莲微量元素含量的测定[J].微量元素与健康研究, 2002, 19(2): 37.
[17]徐建国,任连生. 302种传统治癌中草药水提液对HL-60细胞的诱导分化及细胞毒作用[J].山西医药杂志, 1991, 20(2): 82.
[18] Lee TK, Lee DK, Kim Dl. Inhibitory effeets of Scutellaria barbata D. Don on human uterine leiomyomal smooth muscle cell Proliferation through cell cycle analysis [J]. International Immunopharmacology. 2004, 4(3): 447-454.
[19]李洁,孙静.中药半枝莲对肝癌H22荷瘤小鼠抑瘤作用的研究[J].时珍国医国药, 2009, 20(5): 1233-1234.
[20] Yin x, zhou J, Jie C. Anticaneer activity and mcehanism of Scutellaria barbata extract on humanlun caneer cell Iine A 549[J]. Life Sci, 2004, 75(18): 2233-2244.
[21]周志愉,王兆雷.不同乙醇浓度半枝莲提取物抗肿瘤作用的相关性研究[J].实用中西医结合临床, 2007, 7(6): 89-90.
[22]孟延发,李志孝,王秉峰等.半枝莲多糖的研究[J].兰州大学学报(自然科学版), 1992, 28(4): 112.
[23]曾莉萍,林明宝,舒虹.复方半枝莲胶囊体外抗肿瘤作用的实验研究[J].江西医学院学报, 2006, 46(3): 46-48.
[24]吴东风,林明宝,舒虹等.复方半枝莲胶囊抗肿瘤作用的实验研究[J].实用中西医结合临床, 2006, 6(6): 79-80.
[25]宋寄春,谭诗云,陈明锴.半枝莲对人大肠癌细胞增殖抑制及诱导凋亡的作用[J].咸宁学院学报(医学版) , 2007, 21(2) : 106-108.
[26]韦鹏涯,浦红琴,韦星等.半枝莲提取物诱导人肺癌SPC-A-1细胞凋亡及对凋亡相关基因表达的影响[J].中药材, 2007, 10(30): 1270-1273.
[27]韦鹏涯,浦红琴,韦星等.半枝莲提取物诱导人肝癌SMMC-7721细胞凋亡及其对凋亡相关蛋白表达的影响[J].时珍国医国药, 2007, 18(12): 3020-3022.
[28]谢路琨,邓涛,张秋萍.半枝莲提取物诱导白血病K562细胞凋亡[J].武汉大学学报. 2004, (2): 115-118.
[29]林敬明,刘煜,罗荣城.半枝莲抑制人肝癌QGY-7701细胞增殖研究[J].南方医科大学学报, 2006, 26(5): 591-593.
[30] Yun-Yi Cha, Eun-Ok Lee. Mehtylene chloride fraction of Scutellaria barbata inducesapopsis in human U937 leukemia cells via the miocthnodrial singalnig Phatwya[J]. Clinica Chimica Acta, 2004, 348: 41-48.
[31]段小梅.基因家族对细胞凋亡的调控[J].中国医学文摘肿瘤学, 2001, 15(1): 80.
[32]李洁.半枝莲对人肺巨细胞癌细胞株PG细胞bcl-2基因表达的影响[J].中药药理与临床, 2004, 23(1): 46-47.
[33]高冬,高永琳,白平.半枝莲对宫颈癌细胞钙信号系统的影响[J].中药材, 2003, 26(10): 730-732.
[34]张妮娜,卜平,朱海杭等.半枝莲抑制肿瘤血管生成的作用及其机制研究[J].癌症, 2005, 24(12): 1459-1463.
[35] Wang S, Zheng Z, Weng Y, et a1. Angiogenesis and anti-angiogenesis activity of Chinese medicinal herb al extracts [J]. Life Sci, 2004, 74(20): 2467-2478.
[36]徐敏,卜平,李瑶瑶.半枝莲黄酮类化合物对体外肿瘤血管生长的影响[J].世界华人消化杂志, 2007, 15(20): 2215-2219.
[37] Scambia G, Ferrandina G, Marone M,et a1.nm23 in ovarian cancer:correlation with clinical outcome and other clinicopatho-logie and biochemical prognostic parameters [J]. ClinOncol, 2002, 14( 2): 334.
[38]李洁.半枝莲对人肺巨细胞癌细胞株PG细胞nm23基因表达的影响[J].光明中医, 2008, 23(9): 1262-1263.
[39] Chadeneau C, Hay K, Hirte HW, et al. Telomerase activity associated with acquisition of malignancy in human colorectal cancer[J]. Cancer Res, 1995, 55(12): 2533-2536.
[40]李洁,石俊英.半枝莲对人肺巨细胞癌细胞株PG细胞端粒酶活性的影响[J].中药药理与临床, 2006, 22( 3) : 113-114.
[41]王刚,董玫,刘秀书.半枝莲醇提物抗肿瘤活性的研究[J].现代中西医结合杂志. 2004, (09): 1141-1142.
[42]王洪琦,崔娜娟,胡玲等.清热解毒和补益中药对小鼠腹水肝癌H22细胞的作用及免疫学机制比较[J].广州中医药大学学报, 2006, 23(2): 156-l59.
[43]龙盛京,罗佩卓,覃汗昌. 17种清热中药抗活性氧作用的研究[J].中草药, l999, 30(1): 40-43.
[44]杨容,黄志,习洋等.中药半枝莲中黄酮类化合物的抗氧化活性[J].河北大学学报(自然科学版), 2007, 27(5): 489-491.
[45]贾顺姬,盂安明.利用模式动物探讨出生缺陷的发生机理[J].生命科学, 2006, 18(2): 97-l00.
[46]谢菲,陈晗,李建明等.斑马鱼斑马鱼在肿瘤研究中的应用[J].中国热带医学, 2009, 9(6): 1161-1164.
[47]Folkman J. Tumor angiogenesis: therapentic implications[J]. N Engl J Med. 1971, 285(21): 1182-1186.
[48] laughner E, Taghavi P, Chiles K, et al. HER2 (neu) signaling increases the rate of hypoxia-inducible factors 1a (HIF-1a) synthesis: novel mechanism for HIF-1-Mediated vascular endothelial growth factor expression[J]. Mol Cell Biol. 2001, 21(12): 3995-4004.
[49]CHILDS S, CHEN JN, GARRITY DM, et al. Patterning of Angiogenesis in the Zebrafish Embryo[J]. Development. 2002, 129, 973-982.
[50]夏小艳,刘可春等.大青叶中靛玉红的抗血管生成活性研究[J].中国药学杂志, 2010,45(3):187-189.
[51] Yan-Li Zhao, Qiu-Xia He, Yang Li, Si-Feng Wang, Ke-Chun Liu, Yong-Ping Yang and Xiao-Li Li .Chemical Constituents of Excoecaria acerifolia and Their Bioactivities[J]. Molecules.2010, 15: 2178-2186.
[52] Xiao-Li Li, Yang Li, ?Si-Feng Wang, Yan-Li Zhao,K?e-Chun Liu, Xi-Min Wang, and Yong-Ping Yang. Ingol and Ingenol Diterpenes from the Aerial Parts of Euphorbia royleana and Their Antiangiogenic Activities [J] Nat. Prod. 2009, 7:1001-1005.
[53] Wu Xin-rong, ZhongHan-bing. Study on effective mechanism of mycophenolic acid on antiangiogenesis in flk-GFP zebrafish Chin JCl in Pharmaco[J]. 2008, 24 (4): 311-314.
[54] Chan J, Bayliss P E, Wood J M, et a.l Dissection of angiogenic signaling in zebrafish using a chemical genetic approach[J]. CancerCell. 2002, 1 (3): 257-267.
[55]柳明洙,张学武,高圭永等.血管内皮细胞生长因子和血管发生素-1及其受体在斑马鱼早期胚胎发育过程中的表达[J].时珍国医国药, 2006, 17(6): 912-913.
[56]杨少丽,阎松,郑兰红等.斑马鱼VEGF基因的siRNA表达载体构建、有效序列筛选及其对基因表达的干预性研究[J].生物物理学报, 2007, 23(5): 364-369.
[57] Deepa Alex,Emilia Concei~o Leong, Zai-Jun Zhang等.白藜芦醇衍生物TMS(trans-3,5,4-trimethoxystilbene)在斑马鱼中通过下调VEGFR2和改变细胞周期发挥抗血管新生和血管破坏效应[J].中国药理通讯, 2009, 26(2): 9.
[58]张中朋,杨中林,唐登峰等.地蚕化学成分的分离与鉴定[J].中成药, 2004, 26(12): 1052-1053.
[59]许凤鸣.半枝莲化学的研究[J].中国现代应用药学, 1997, 14(6): 8-9.
[60] M.Grande, F.Piera, A.Cuenca et. al. Flavonoids from Inula viscosa[J]. Plant Medical, 1985, 414-417.
[61]徐燕,张正竹,邹忠梅.香附的化学成分研究[J].中国药学杂志, 2010, 5 (1): 818-821.
[62]丁林芬,郭亚东,吴兴德等.珍珠菜黄酮类化学成分研究[J].中成药, 2010, 32(5): 827-831.
[63]邵帅,严铭铭,毕胜男等.小飞蓬化学成分的研究[J].中国药学杂志, 2009, 44(14): 1053-1055.
[64]吴家明,陆茵,郜明等.血管生成实验模型研究进展[J].中国药理学通报,2008,24(1): 11-14.
[65] Kidd K R, Weinstein B M. Fishing for novel angiogenic therapies. British Journal of Pharmacology[J]. 2003, 140 (4): 585-594.
[66] Lawson N D, Weinstein B M. In vivo imaging of embryonic vascular development using transgenic zebrafish[J]. Dev Biol, 2002, 248 (2): 307-318.
[67] Matz M V, Fradkov A F, Labas Y A, et al. Fluorescent proteins from nonbioluminescent Anthozoa species[J]. Nat Biotech, 2000, 17(10): 969-973.
[68] Hou J M, Liu J Y, Yang L, et al . Combination of low-do se gemcitabine and recom binant quail vascular endothelial growth factor receptor 2 as a vaccine induces synergistic antitumor activities[J].Oncology, 2005, 69(1): 81-87.
[69]陈维,章茂顺,王家良等.槲皮素及异鼠李素对人血管平滑肌细胞胶原合成的影响[J].中国动脉硬化杂志, 2005, 13(3): 320-324.
[70] Bao M, Lou Y. Isorhamnetin prevent endothe lial cell injuries from oxidized LDL via activation of p38 MAPK[J]. EurJ Pharmacol, 2006, 547(1/3): 222-230.
[71] Igura k ohta T kuroda Y et al. Resveratrol and quercetin inhibit angiogenesis in Vitro. Cancer Lett[J], 2001, 171(1): 11-16.
[72]王晓庆,梁中琴,顾振纶等槲皮素抑制血管生成作用的实验研究[J].中国药理学通报, 2004, 20 (10): 1161-1164.
[73] Fan PS, Gu ZL,Sheng R etal. Inhibitory effect of quercetin on prolif 2 ration of human microvascular endothelial cells in vitro [J]. Acta Pharmacologica Sinica, 2003, 24 (12): 1231-1234.
[74].钟晓刚.槲皮素对乳腺癌血管生成、增殖和凋亡的影响[D].苏州:苏州大学硕士论文, 2004.
[2]国家药典委员会.中华人民共和国药典(一部) [M]北京:化学工业出版社, 2005, 77.
[3]蒋小刚.半枝莲的化学成分和药理作用[J].中国野生植物资源,2004,23(1):4-6.
[4] Lin Y-1, Chou C-J. Studies on the constituents of acrial parts of Sutslaria rivularis Wall[J]. Kou Li Chung-Kuo YaoYenChiu SoYen Chiu Pao Kao, 1984, (7), 141-143.
[5] Chou Cheng jen, Liu Kao Chu, Yang Tsang Hsiung. The isolation and characterization of scutevulin, and some other constituents from Scutellaria rivularis wall[D].Guoli Zhongguo Yiyao Yangjiuso Yanjiu Baogao, 1981 :91-108.
[6]向仁德,郑今,姚志成.半枝莲化学成分的研究[J].中草药,1982,13(8),9-12.
[7] Sonoda M, Nishiyama T, Matsukwa Y. Cytotoxic aetivities of flavonoids From two Seutellaria plants in Chinese medieine [J]. Ethnopharmeol, 2004, 91(1): 65-68
[8] Tomimori T. Chem Pharm Bull[D]. 1990, 38 (12), 3488.
[9]杨顺利,方岩雄.中药半枝莲抗肿瘤成分的分离研究[D].广州工业大学, 2002: 26-38.
[10]许益民,郭立伟,陈建伟.半枝莲多糖的分离纯化及其理化性质[J].天然产物研究与开发, 1992, 4(l): 1-5.
[11]孟研发,李志孝,张立等.半枝莲多糖SPS4组分的纯化性质及分析[J].生物化学杂志, 1993, 9(2): 224-228.
[12] Wang ZQ, Xu FM, Yan XZ, et al. Scutebarbatine A, a new neoelerodane type diterpeonid alkaloid from Seutellaria barbata[J]. Chin.Chem.Lett, 1996, 7(4): 333-334.
[13] Sheng JD, Jia YT, KeL, et al. Neo-elerodane type diterpenoides from Scutellaria barbata with Cytotoxic activities [J]. Phytoehemistry, 2006, 67: 1326-1330.
[14] Dai SJ, Chen M, Liu K, et al. Four New neo-Clerodane Diterpenoid Alkaloids from Scutellaria barbata with Cytotoxiec Aetivities [J]. Chem Pharm Bull, 2006, 54(6): 869-872.
[15] Ducki S, Hadfield JA, Lawrence NJ, et al.Isolation of E-1(4′ydroxyphenyl)-but-1-en-3-one from Scutellaria barbata[J]. Planta Med, 1996, 62(2): 185-186.
[16]席荣英,李聪辉,王翠红.半枝莲微量元素含量的测定[J].微量元素与健康研究, 2002, 19(2): 37.
[17]徐建国,任连生. 302种传统治癌中草药水提液对HL-60细胞的诱导分化及细胞毒作用[J].山西医药杂志, 1991, 20(2): 82.
[18] Lee TK, Lee DK, Kim Dl. Inhibitory effeets of Scutellaria barbata D. Don on human uterine leiomyomal smooth muscle cell Proliferation through cell cycle analysis [J]. International Immunopharmacology. 2004, 4(3): 447-454.
[19]李洁,孙静.中药半枝莲对肝癌H22荷瘤小鼠抑瘤作用的研究[J].时珍国医国药, 2009, 20(5): 1233-1234.
[20] Yin x, zhou J, Jie C. Anticaneer activity and mcehanism of Scutellaria barbata extract on humanlun caneer cell Iine A 549[J]. Life Sci, 2004, 75(18): 2233-2244.
[21]周志愉,王兆雷.不同乙醇浓度半枝莲提取物抗肿瘤作用的相关性研究[J].实用中西医结合临床, 2007, 7(6): 89-90.
[22]孟延发,李志孝,王秉峰等.半枝莲多糖的研究[J].兰州大学学报(自然科学版), 1992, 28(4): 112.
[23]曾莉萍,林明宝,舒虹.复方半枝莲胶囊体外抗肿瘤作用的实验研究[J].江西医学院学报, 2006, 46(3): 46-48.
[24]吴东风,林明宝,舒虹等.复方半枝莲胶囊抗肿瘤作用的实验研究[J].实用中西医结合临床, 2006, 6(6): 79-80.
[25]宋寄春,谭诗云,陈明锴.半枝莲对人大肠癌细胞增殖抑制及诱导凋亡的作用[J].咸宁学院学报(医学版) , 2007, 21(2) : 106-108.
[26]韦鹏涯,浦红琴,韦星等.半枝莲提取物诱导人肺癌SPC-A-1细胞凋亡及对凋亡相关基因表达的影响[J].中药材, 2007, 10(30): 1270-1273.
[27]韦鹏涯,浦红琴,韦星等.半枝莲提取物诱导人肝癌SMMC-7721细胞凋亡及其对凋亡相关蛋白表达的影响[J].时珍国医国药, 2007, 18(12): 3020-3022.
[28]谢路琨,邓涛,张秋萍.半枝莲提取物诱导白血病K562细胞凋亡[J].武汉大学学报. 2004, (2): 115-118.
[29]林敬明,刘煜,罗荣城.半枝莲抑制人肝癌QGY-7701细胞增殖研究[J].南方医科大学学报, 2006, 26(5): 591-593.
[30] Yun-Yi Cha, Eun-Ok Lee. Mehtylene chloride fraction of Scutellaria barbata inducesapopsis in human U937 leukemia cells via the miocthnodrial singalnig Phatwya[J]. Clinica Chimica Acta, 2004, 348: 41-48.
[31]段小梅.基因家族对细胞凋亡的调控[J].中国医学文摘肿瘤学, 2001, 15(1): 80.
[32]李洁.半枝莲对人肺巨细胞癌细胞株PG细胞bcl-2基因表达的影响[J].中药药理与临床, 2004, 23(1): 46-47.
[33]高冬,高永琳,白平.半枝莲对宫颈癌细胞钙信号系统的影响[J].中药材, 2003, 26(10): 730-732.
[34]张妮娜,卜平,朱海杭等.半枝莲抑制肿瘤血管生成的作用及其机制研究[J].癌症, 2005, 24(12): 1459-1463.
[35] Wang S, Zheng Z, Weng Y, et a1. Angiogenesis and anti-angiogenesis activity of Chinese medicinal herb al extracts [J]. Life Sci, 2004, 74(20): 2467-2478.
[36]徐敏,卜平,李瑶瑶.半枝莲黄酮类化合物对体外肿瘤血管生长的影响[J].世界华人消化杂志, 2007, 15(20): 2215-2219.
[37] Scambia G, Ferrandina G, Marone M,et a1.nm23 in ovarian cancer:correlation with clinical outcome and other clinicopatho-logie and biochemical prognostic parameters [J]. ClinOncol, 2002, 14( 2): 334.
[38]李洁.半枝莲对人肺巨细胞癌细胞株PG细胞nm23基因表达的影响[J].光明中医, 2008, 23(9): 1262-1263.
[39] Chadeneau C, Hay K, Hirte HW, et al. Telomerase activity associated with acquisition of malignancy in human colorectal cancer[J]. Cancer Res, 1995, 55(12): 2533-2536.
[40]李洁,石俊英.半枝莲对人肺巨细胞癌细胞株PG细胞端粒酶活性的影响[J].中药药理与临床, 2006, 22( 3) : 113-114.
[41]王刚,董玫,刘秀书.半枝莲醇提物抗肿瘤活性的研究[J].现代中西医结合杂志. 2004, (09): 1141-1142.
[42]王洪琦,崔娜娟,胡玲等.清热解毒和补益中药对小鼠腹水肝癌H22细胞的作用及免疫学机制比较[J].广州中医药大学学报, 2006, 23(2): 156-l59.
[43]龙盛京,罗佩卓,覃汗昌. 17种清热中药抗活性氧作用的研究[J].中草药, l999, 30(1): 40-43.
[44]杨容,黄志,习洋等.中药半枝莲中黄酮类化合物的抗氧化活性[J].河北大学学报(自然科学版), 2007, 27(5): 489-491.
[45]贾顺姬,盂安明.利用模式动物探讨出生缺陷的发生机理[J].生命科学, 2006, 18(2): 97-l00.
[46]谢菲,陈晗,李建明等.斑马鱼斑马鱼在肿瘤研究中的应用[J].中国热带医学, 2009, 9(6): 1161-1164.
[47]Folkman J. Tumor angiogenesis: therapentic implications[J]. N Engl J Med. 1971, 285(21): 1182-1186.
[48] laughner E, Taghavi P, Chiles K, et al. HER2 (neu) signaling increases the rate of hypoxia-inducible factors 1a (HIF-1a) synthesis: novel mechanism for HIF-1-Mediated vascular endothelial growth factor expression[J]. Mol Cell Biol. 2001, 21(12): 3995-4004.
[49]CHILDS S, CHEN JN, GARRITY DM, et al. Patterning of Angiogenesis in the Zebrafish Embryo[J]. Development. 2002, 129, 973-982.
[50]夏小艳,刘可春等.大青叶中靛玉红的抗血管生成活性研究[J].中国药学杂志, 2010,45(3):187-189.
[51] Yan-Li Zhao, Qiu-Xia He, Yang Li, Si-Feng Wang, Ke-Chun Liu, Yong-Ping Yang and Xiao-Li Li .Chemical Constituents of Excoecaria acerifolia and Their Bioactivities[J]. Molecules.2010, 15: 2178-2186.
[52] Xiao-Li Li, Yang Li, ?Si-Feng Wang, Yan-Li Zhao,K?e-Chun Liu, Xi-Min Wang, and Yong-Ping Yang. Ingol and Ingenol Diterpenes from the Aerial Parts of Euphorbia royleana and Their Antiangiogenic Activities [J] Nat. Prod. 2009, 7:1001-1005.
[53] Wu Xin-rong, ZhongHan-bing. Study on effective mechanism of mycophenolic acid on antiangiogenesis in flk-GFP zebrafish Chin JCl in Pharmaco[J]. 2008, 24 (4): 311-314.
[54] Chan J, Bayliss P E, Wood J M, et a.l Dissection of angiogenic signaling in zebrafish using a chemical genetic approach[J]. CancerCell. 2002, 1 (3): 257-267.
[55]柳明洙,张学武,高圭永等.血管内皮细胞生长因子和血管发生素-1及其受体在斑马鱼早期胚胎发育过程中的表达[J].时珍国医国药, 2006, 17(6): 912-913.
[56]杨少丽,阎松,郑兰红等.斑马鱼VEGF基因的siRNA表达载体构建、有效序列筛选及其对基因表达的干预性研究[J].生物物理学报, 2007, 23(5): 364-369.
[57] Deepa Alex,Emilia Concei~o Leong, Zai-Jun Zhang等.白藜芦醇衍生物TMS(trans-3,5,4-trimethoxystilbene)在斑马鱼中通过下调VEGFR2和改变细胞周期发挥抗血管新生和血管破坏效应[J].中国药理通讯, 2009, 26(2): 9.
[58]张中朋,杨中林,唐登峰等.地蚕化学成分的分离与鉴定[J].中成药, 2004, 26(12): 1052-1053.
[59]许凤鸣.半枝莲化学的研究[J].中国现代应用药学, 1997, 14(6): 8-9.
[60] M.Grande, F.Piera, A.Cuenca et. al. Flavonoids from Inula viscosa[J]. Plant Medical, 1985, 414-417.
[61]徐燕,张正竹,邹忠梅.香附的化学成分研究[J].中国药学杂志, 2010, 5 (1): 818-821.
[62]丁林芬,郭亚东,吴兴德等.珍珠菜黄酮类化学成分研究[J].中成药, 2010, 32(5): 827-831.
[63]邵帅,严铭铭,毕胜男等.小飞蓬化学成分的研究[J].中国药学杂志, 2009, 44(14): 1053-1055.
[64]吴家明,陆茵,郜明等.血管生成实验模型研究进展[J].中国药理学通报,2008,24(1): 11-14.
[65] Kidd K R, Weinstein B M. Fishing for novel angiogenic therapies. British Journal of Pharmacology[J]. 2003, 140 (4): 585-594.
[66] Lawson N D, Weinstein B M. In vivo imaging of embryonic vascular development using transgenic zebrafish[J]. Dev Biol, 2002, 248 (2): 307-318.
[67] Matz M V, Fradkov A F, Labas Y A, et al. Fluorescent proteins from nonbioluminescent Anthozoa species[J]. Nat Biotech, 2000, 17(10): 969-973.
[68] Hou J M, Liu J Y, Yang L, et al . Combination of low-do se gemcitabine and recom binant quail vascular endothelial growth factor receptor 2 as a vaccine induces synergistic antitumor activities[J].Oncology, 2005, 69(1): 81-87.
[69]陈维,章茂顺,王家良等.槲皮素及异鼠李素对人血管平滑肌细胞胶原合成的影响[J].中国动脉硬化杂志, 2005, 13(3): 320-324.
[70] Bao M, Lou Y. Isorhamnetin prevent endothe lial cell injuries from oxidized LDL via activation of p38 MAPK[J]. EurJ Pharmacol, 2006, 547(1/3): 222-230.
[71] Igura k ohta T kuroda Y et al. Resveratrol and quercetin inhibit angiogenesis in Vitro. Cancer Lett[J], 2001, 171(1): 11-16.
[72]王晓庆,梁中琴,顾振纶等槲皮素抑制血管生成作用的实验研究[J].中国药理学通报, 2004, 20 (10): 1161-1164.
[73] Fan PS, Gu ZL,Sheng R etal. Inhibitory effect of quercetin on prolif 2 ration of human microvascular endothelial cells in vitro [J]. Acta Pharmacologica Sinica, 2003, 24 (12): 1231-1234.
[74].钟晓刚.槲皮素对乳腺癌血管生成、增殖和凋亡的影响[D].苏州:苏州大学硕士论文, 2004.