16种中药水提物和醇提物对黄嘌呤氧化酶的抑制作用
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摘要
为了筛选治疗禽痛风病的中药,试验对16种中药进行了水提和醇提,并检测提取物对黄嘌呤氧化酶(XOD)活性的体外抑制作用,采用紫外分光光度法确定反应体系的检测波长、酶浓度、检测时间,并检测同一水提和醇提条件下的16种中药提取物对XOD的抑制率,测定每种中药的半数抑制浓度(IC_(50))。结果表明:检测波长为292 nm,酶浓度为34.0μg/mL,检测时间为第1,2,3,4,5,6,7,8,9,10分钟;同一种中药的水提物和醇提物对XOD的抑制作用存在一定差异;中药水提物中抑制效果较好的有黄柏、土茯苓、槐花、高良姜、山慈菇,其IC_(50)分别为12.60,15.98,19.66,29.65,31.68 mg/mL;中药醇提物中抑制效果较好的有土茯苓、瞿麦、槐花、牛膝、山慈菇,其IC_(50)分别为14.89,16.99,23.44,29.48,29.61 mg/mL。说明同一种中药水提物和醇提物对XOD抑制作用存在一定差异。
In order to screen the traditional Chinese medicines for treating poultry gout, water and alcohol extraction were carried out on 16 kinds of traditional Chinese medicines, and the in vitro inhibition effect of them on xanthine oxidase(XOD) activity was detected.UV spectrophotometry was used to determine the wavelength, enzyme concentration and time of the reaction system. The inhibition rates of 16 Chinese herbal extracts on XOD under the same water extraction and alcohol extraction condition were determined, and the 50% inhibition concentrations of each Chinese herbal medicine(IC_(50)) were detected.The results showed that the wavelength of detection was 292 nm, the enzyme concentration was 34.0 μg/mL, and the detection time were 1,2,3,4,5,6,7,8,9,10 minutes.There were differences between the water extraction and ethanol extraction of each Chinese medicine.The better inhibitory effective water extracts were Phellodendri Chinensis Cortex, Glabrous Greenbrier Rhizome,Sophorae Flos,Alpinia officinarum hance,Sagittaria.The IC_(50) were 12.60, 15.98, 19.66, 29.65, 31.68 mg/mL, respectively.The better inhibitory effective alcohol extracts were Glabrous Greenbrier Rhizome, Dianthus Superbus L, Sophorae Flos,Achyranthes bidentata, Sagittaria,the IC_(50) were 14.89,16.99,23.44,29.48,29.61 mg/mL, respectively. It shows that there are some differences between the same Chinese medicine water extract and alcohol extract on the inhibition of XOD.
In order to screen the traditional Chinese medicines for treating poultry gout, water and alcohol extraction were carried out on 16 kinds of traditional Chinese medicines, and the in vitro inhibition effect of them on xanthine oxidase(XOD) activity was detected.UV spectrophotometry was used to determine the wavelength, enzyme concentration and time of the reaction system. The inhibition rates of 16 Chinese herbal extracts on XOD under the same water extraction and alcohol extraction condition were determined, and the 50% inhibition concentrations of each Chinese herbal medicine(IC_(50)) were detected.The results showed that the wavelength of detection was 292 nm, the enzyme concentration was 34.0 μg/mL, and the detection time were 1,2,3,4,5,6,7,8,9,10 minutes.There were differences between the water extraction and ethanol extraction of each Chinese medicine.The better inhibitory effective water extracts were Phellodendri Chinensis Cortex, Glabrous Greenbrier Rhizome,Sophorae Flos,Alpinia officinarum hance,Sagittaria.The IC_(50) were 12.60, 15.98, 19.66, 29.65, 31.68 mg/mL, respectively.The better inhibitory effective alcohol extracts were Glabrous Greenbrier Rhizome, Dianthus Superbus L, Sophorae Flos,Achyranthes bidentata, Sagittaria,the IC_(50) were 14.89,16.99,23.44,29.48,29.61 mg/mL, respectively. It shows that there are some differences between the same Chinese medicine water extract and alcohol extract on the inhibition of XOD.
引文
[1] 黄克和,唐建霞.禽痛风研究现状[J].中国兽医杂志,2005, 41(5):30-32.
[2] LIN H, CHEN Y, HUANG Q, et al. Prokaryotic expression of the chicken xanthine oxidase (XOD) subunit and its localization in liver and kidney[J]. Int J Biol Macromol, 2016, 87:341-347.
[3] 陆海波,鲁传华.高尿酸血症治疗药物黄嘌呤氧化酶抑制剂的研究进展[J].安徽医药,2016,20(4):626-630.
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[5] SHEU S Y,LIN Y C,CHIANG H C. Inhibition of xanthine oxidase by synthetic cytokinin analogues[J]. Anticancer Res, 1997, 17(2A):1043-1049.
[6] GUPTA S,RODRIGUES L M,ESTEVES A P, et al. Synthesis of N-aryl-5-amino-4-cyanopyrazole derivatives as potent xanthine oxidase inhibitors[J]. Eur J Med Chem, 2008, 43(4):771-780.
[7] COS P,YING L,CALOMME M, et al. Structure-activity relationship and classification of flavonoids as inhibitors of xanthine oxidase and superoxide scavengers[J]. J Nat Prod, 1998, 61(1):71-76.
[8] 申启荣.中药黄嘌呤氧化酶抑制剂的筛选及抑制动力学研究[D].南昌:南昌大学,2015.
[9] 雷鹏,郭小权,曹华斌,等.禽痛风的研究进展[J].中国家禽,2011,33(2):48-51.
[10] 王晓梅, 赵岩, 刘丹,等. 三种中药提取物对黄嘌呤氧化酶的抑制作用[J]. 氨基酸和生物资源, 2013, 35(1):57-60.
[11] 黎莉,戴立珍,杨靖,等.中药提取物中黄嘌呤氧化酶抑制剂的筛选[J].武汉工程大学学报, 2010, 32(3):44-46.
[12] 许洪波,周瑞,谢培,等.27种中草药的黄嘌呤氧化酶抑制活性筛选[J].时珍国医国药,2017(3):547-548.
[13] 王小龙.畜禽营养代谢病和中毒病[M]. 北京:中国农业出版社,2003.
[14] 朱春胜, 张冰, 林志健,等.中医药治疗高尿酸血症的研究进展[J].中华中医药杂志, 2015(12):4374-4376.
[15] 谢宗万.全国中草药汇编[M].北京:人民卫生出版社,1996.
[16] 朱立然.萆薢总皂苷对高尿酸血症大鼠尿酸转运蛋白表达的影响[D].合肥:安徽中医药大学,2013.
[2] LIN H, CHEN Y, HUANG Q, et al. Prokaryotic expression of the chicken xanthine oxidase (XOD) subunit and its localization in liver and kidney[J]. Int J Biol Macromol, 2016, 87:341-347.
[3] 陆海波,鲁传华.高尿酸血症治疗药物黄嘌呤氧化酶抑制剂的研究进展[J].安徽医药,2016,20(4):626-630.
[4] HOORN D E C V, NIJVELDT R J, LEEUWEN PAMV, et al. Accurate prediction of xanthine oxidase inhibition based on the structure of flavonoids[J]. Eur J Pharmacol, 2002, 451(2):111-118.
[5] SHEU S Y,LIN Y C,CHIANG H C. Inhibition of xanthine oxidase by synthetic cytokinin analogues[J]. Anticancer Res, 1997, 17(2A):1043-1049.
[6] GUPTA S,RODRIGUES L M,ESTEVES A P, et al. Synthesis of N-aryl-5-amino-4-cyanopyrazole derivatives as potent xanthine oxidase inhibitors[J]. Eur J Med Chem, 2008, 43(4):771-780.
[7] COS P,YING L,CALOMME M, et al. Structure-activity relationship and classification of flavonoids as inhibitors of xanthine oxidase and superoxide scavengers[J]. J Nat Prod, 1998, 61(1):71-76.
[8] 申启荣.中药黄嘌呤氧化酶抑制剂的筛选及抑制动力学研究[D].南昌:南昌大学,2015.
[9] 雷鹏,郭小权,曹华斌,等.禽痛风的研究进展[J].中国家禽,2011,33(2):48-51.
[10] 王晓梅, 赵岩, 刘丹,等. 三种中药提取物对黄嘌呤氧化酶的抑制作用[J]. 氨基酸和生物资源, 2013, 35(1):57-60.
[11] 黎莉,戴立珍,杨靖,等.中药提取物中黄嘌呤氧化酶抑制剂的筛选[J].武汉工程大学学报, 2010, 32(3):44-46.
[12] 许洪波,周瑞,谢培,等.27种中草药的黄嘌呤氧化酶抑制活性筛选[J].时珍国医国药,2017(3):547-548.
[13] 王小龙.畜禽营养代谢病和中毒病[M]. 北京:中国农业出版社,2003.
[14] 朱春胜, 张冰, 林志健,等.中医药治疗高尿酸血症的研究进展[J].中华中医药杂志, 2015(12):4374-4376.
[15] 谢宗万.全国中草药汇编[M].北京:人民卫生出版社,1996.
[16] 朱立然.萆薢总皂苷对高尿酸血症大鼠尿酸转运蛋白表达的影响[D].合肥:安徽中医药大学,2013.