不同提取工艺对炒决明子急性毒性和物质基础的影响
|
摘要
目的研究不同提取工艺对炒决明子急性毒性和物质基础的影响,为炒决明子的临床安全用药提供依据。方法分别制备炒决明子水提物和醇提物,用高效液相色谱法测定大黄酚和橙黄决明素含量。预试验结果表明由于受给药体积及浓度的限制,难以找出炒决明子水提物组和醇提物对小鼠的半数致死量(LD50),进行最大耐受量(MTD)或最大给药量(MFD)的测定。正常小鼠按性别、体质量随机分成3组:对照组、炒决明子水提物组、炒决明子醇提物组,每组20只。水提物组按40.0 mL/kg给药1次,12 h后无死亡情况,进行第2次给药。醇提物组按40.0 mL/kg给药1次,12 h内出现死亡情况,不进行第2次给药,对照组小鼠ig相同体积的水。观察小鼠体质量、饮食、饮水,14 d内可能出现的呼吸频率改变、呼吸困难、动度、对外反应情况、排便异常等,检测血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、肌酐(Cr)、尿素氮(BUN)水平。结果制备的炒决明子水提物按生药量计为0.952 g/mL,大黄酚、橙黄决明素分别占生药的0.145 1%、0.018 5%;制备的炒决明子醇提物按生药量计为3.950 g/mL,大黄酚、橙黄决明素分别占生药的0.144 9%、0.024 8%。炒决明子水提物对正常小鼠的MFD为114.96 g/kg,相当于临床成年人日用量的588倍;炒决明子醇提物对正常小鼠MTD为178.40 g/kg,相当于临床成年人日用量的832.5倍。炒决明子水、醇提取物的急毒主要表现为:出现间歇扭体反应、便溏、呼吸频率减慢、呼吸困难、精神萎靡、俯卧昏睡等,其中便溏现象尤为明显,且醇提物>水提物。给药后,炒决明子水提物组和醇提物组小鼠的ALT、AST、BUN、Cr指标升高,与对照组比较,水提物组ALT、 BUN、 Cr水平呈显著性差异(P<0.01、0.001),AST未见显著性差异;醇提物组ALT、BUN、Cr水平呈显著性差异(P<0.001),AST未呈现显著性差异。结论大剂量炒决明子水、醇提物对正常小鼠的肝、肾功能均有影响,其中对肾脏的影响较为严重;临床应用中,尽量选用炒决明子水提工艺以保证安全有效。
Objective To study the relationship between acute toxicity and material basis of the roasted seeds of Cassia obtusifolia by different extraction techniques, and provide a basis for safe clinical use. Methods The water extract and alcohol extract of the roasted seeds of C. obtusifolia was prepared, the content of chrysophanol and aurantio-obtusin in different extracts was determinated by HPLC. Pre-test results showed that it was difficult to find out the LD50 of water extract and alcohol extract in mice due to the limitation of drug volume and concentration, the maximum tolerable dose(MTD) or the maximum dose(MFD) were determined.Normal mice were randomly divided into three groups according to sex and body weight: control group, water extract group and alcohol extract group, with 20 mice in each group. In the water extract group, 40.0 mL/kg was administered once, and no death occurred 12 h later, and the second dose was administered. The ethanol extract group was given 40.0 mL/kg once, and death occurred within 12 h, and no second dose was given. The mice in the control group were received the same volume of water by ig.The changes of body weight, diet, drinking water, respiratory frequency, dyspnea, motility, external reaction and abnormal defecation were observed within 14 d. The levels of serum aspartate aminotransferase(AST), alanine aminotransferase(ALT),creatinine(Cr), urea nitrogen(BUN) were detected. Results The water extract was 0.952 g/mL according to the amount of crude drugs, chrysophanol and aurantio-obtusin accounted for 0.1451% and 0.0185% in water extract, respectively; The alcohol extract was 3.950 g/mL according to the amount of crude drugs, chrysophanol accounted for 0.1449%, aurantio-obtusin accounted for0.0248%; The MFD of the roasted seeds of C. obtusifolia water extract was 114.96 g/kg for normal mice, which was 588 times of the daily dosage per kilogram of body weight of 70 kg adults. The MTD of roasted seeds of C. obtusifolia ethanol extract was 178.40 g/kgfor normal mice, which was 832.5 times the daily dosage per kilogram of body weight of 70 kg adults. The acute poison of water and alcohol extract of the roasted seeds of C. obtusifolia was mainly manifested as follows: intermittent twisting reaction,loose stool, slow breathing rate, dyspnea, depression, prone to lethargy, etc,the phenomenon of loose stool was particularly obvious,and ethanol extract > water extract. The ALT, BUN, Cr levels increased in aqueous extract group and alcohol extract group.Compared with control group, the aqueous extract group ALT, BUN, Cr levels show very significant difference(P < 0.01 and 0.001),AST did not show significant difference. The ALT, BUN and Cr levels of alcohol extract group were supremely significant different(P < 0.001), AST was not significantly different. Conclusion High dose of water and alcohol extracts of the roasted seeds of C.obtusifolia have effects on liver and kidney function of normal mice, among which the effects on kidney are more serious. In clinical application, the water extraction process of the roasted seeds of C. obtusifolia should be selected as far as possible to ensure its safety and effectiveness.
Objective To study the relationship between acute toxicity and material basis of the roasted seeds of Cassia obtusifolia by different extraction techniques, and provide a basis for safe clinical use. Methods The water extract and alcohol extract of the roasted seeds of C. obtusifolia was prepared, the content of chrysophanol and aurantio-obtusin in different extracts was determinated by HPLC. Pre-test results showed that it was difficult to find out the LD50 of water extract and alcohol extract in mice due to the limitation of drug volume and concentration, the maximum tolerable dose(MTD) or the maximum dose(MFD) were determined.Normal mice were randomly divided into three groups according to sex and body weight: control group, water extract group and alcohol extract group, with 20 mice in each group. In the water extract group, 40.0 mL/kg was administered once, and no death occurred 12 h later, and the second dose was administered. The ethanol extract group was given 40.0 mL/kg once, and death occurred within 12 h, and no second dose was given. The mice in the control group were received the same volume of water by ig.The changes of body weight, diet, drinking water, respiratory frequency, dyspnea, motility, external reaction and abnormal defecation were observed within 14 d. The levels of serum aspartate aminotransferase(AST), alanine aminotransferase(ALT),creatinine(Cr), urea nitrogen(BUN) were detected. Results The water extract was 0.952 g/mL according to the amount of crude drugs, chrysophanol and aurantio-obtusin accounted for 0.1451% and 0.0185% in water extract, respectively; The alcohol extract was 3.950 g/mL according to the amount of crude drugs, chrysophanol accounted for 0.1449%, aurantio-obtusin accounted for0.0248%; The MFD of the roasted seeds of C. obtusifolia water extract was 114.96 g/kg for normal mice, which was 588 times of the daily dosage per kilogram of body weight of 70 kg adults. The MTD of roasted seeds of C. obtusifolia ethanol extract was 178.40 g/kgfor normal mice, which was 832.5 times the daily dosage per kilogram of body weight of 70 kg adults. The acute poison of water and alcohol extract of the roasted seeds of C. obtusifolia was mainly manifested as follows: intermittent twisting reaction,loose stool, slow breathing rate, dyspnea, depression, prone to lethargy, etc,the phenomenon of loose stool was particularly obvious,and ethanol extract > water extract. The ALT, BUN, Cr levels increased in aqueous extract group and alcohol extract group.Compared with control group, the aqueous extract group ALT, BUN, Cr levels show very significant difference(P < 0.01 and 0.001),AST did not show significant difference. The ALT, BUN and Cr levels of alcohol extract group were supremely significant different(P < 0.001), AST was not significantly different. Conclusion High dose of water and alcohol extracts of the roasted seeds of C.obtusifolia have effects on liver and kidney function of normal mice, among which the effects on kidney are more serious. In clinical application, the water extraction process of the roasted seeds of C. obtusifolia should be selected as far as possible to ensure its safety and effectiveness.
引文
[1]中国药典[S].一部. 2015.
[2]黄娜娜,孙蓉.决明子不同提取方式对正常小鼠的急性毒性比较研究[J].中国药物警戒, 2016, 13(11):641-644, 650.
[3]徐叔云,卞如濂.药理实验方法学[M].北京:人民卫生出版社, 2002:227-229.
[4]张加雄,万丽,胡轶娟,等.决明子降血脂有效部位的研究[J].时珍国医国药, 2006, 17(6):904-905.
[5]刘伟民,江滨,曾元儿,等.决明子总蒽醌的水提和醇提工艺比较研究[J].时珍国医国药, 2009, 20(9):2270-2271.
[6]项昭保,金光辉,王光利.决明子有效成分提取工艺和药理作用研究进展[J].食品工业科技, 2013, 34(2):387-391.
[7]尚冀宁.决明子化学成分及提取方法研究[J].科技创新导报, 2013,(20):134-135.
[8]张治雄,梁永锋.决明子化学成分的分离与鉴定[J].中国药房, 2012, 23(19):1782-1783.
[9]黄娜娜,窦立雯,孙蓉.基于功效及物质基础的决明子药理与毒理作用研究进展[J].中国药物警戒, 2016,13(5):282-285.
[10]李桂柳,肖永庆,李丽,等.炒决明子化学成分的研究[J].中国中药杂志, 2009, 34(1):54-56.
[11]高钦,许惠琴,陈建伟,等.不同炮制的决明子保肝及润肠通便作用研究[J].中药新药与临床药理, 2007,(3):194-196.
[12]李桂柳,肖永庆,张村,等.决明子炒制前后2类成分的含量比较研究[J].中国中药杂志, 2009, 34(11):1364-1367.
[13]赵岩.决明子炮制前后化学成分变化研究[J].时珍国医国药, 2010, 21(10):2516-2518.
[2]黄娜娜,孙蓉.决明子不同提取方式对正常小鼠的急性毒性比较研究[J].中国药物警戒, 2016, 13(11):641-644, 650.
[3]徐叔云,卞如濂.药理实验方法学[M].北京:人民卫生出版社, 2002:227-229.
[4]张加雄,万丽,胡轶娟,等.决明子降血脂有效部位的研究[J].时珍国医国药, 2006, 17(6):904-905.
[5]刘伟民,江滨,曾元儿,等.决明子总蒽醌的水提和醇提工艺比较研究[J].时珍国医国药, 2009, 20(9):2270-2271.
[6]项昭保,金光辉,王光利.决明子有效成分提取工艺和药理作用研究进展[J].食品工业科技, 2013, 34(2):387-391.
[7]尚冀宁.决明子化学成分及提取方法研究[J].科技创新导报, 2013,(20):134-135.
[8]张治雄,梁永锋.决明子化学成分的分离与鉴定[J].中国药房, 2012, 23(19):1782-1783.
[9]黄娜娜,窦立雯,孙蓉.基于功效及物质基础的决明子药理与毒理作用研究进展[J].中国药物警戒, 2016,13(5):282-285.
[10]李桂柳,肖永庆,李丽,等.炒决明子化学成分的研究[J].中国中药杂志, 2009, 34(1):54-56.
[11]高钦,许惠琴,陈建伟,等.不同炮制的决明子保肝及润肠通便作用研究[J].中药新药与临床药理, 2007,(3):194-196.
[12]李桂柳,肖永庆,张村,等.决明子炒制前后2类成分的含量比较研究[J].中国中药杂志, 2009, 34(11):1364-1367.
[13]赵岩.决明子炮制前后化学成分变化研究[J].时珍国医国药, 2010, 21(10):2516-2518.