乌腺金丝桃中金丝桃素超声提取工艺及其抗病毒性能
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摘要
以金丝桃素提取量为评价指标,在单因素实验基础上,采用响应面法对提取因素进行优化,建立了超声提取乌腺金丝桃中金丝桃素的工艺数学模型。确定最佳提取条件为:提取温度为41℃,丙酮体积分数为69.5%,提取时间为43.5 min,提取溶剂体积(m L)与药材质量(g)比值即液料比为15.3,在该条件下金丝桃素提取量可达4.70 mg/g。主效应关系为提取时间>丙酮浓度>提取温度>液料比。采用细胞病变效应(CPE)法检测乌腺金丝桃提取物体外抗传染性支气管炎病毒(IBV)作用。乌腺金丝桃提取物在鸡胚肾细胞(CEK)感染IBV前、后及同时加入,有一定抑制细胞病变的作用,说明乌腺金丝桃提取物具有一定的抗IBV作用。
Based on the single factor experiment, the response surface method was used to optimize the key extraction factors affecting ultrasonic extraction of hypericin from Hypericum attenuatum Choisy with the extraction amount of hypericin as evaluation index and a mathematical model was established. The optimum conditions for ultrasonic extraction of hypericin were as follows: extraction temperature 41 ℃,acetone volume fraction 69.5%, extraction time 43.5 min, liquid to material ratio 15.3. Under these conditions, the extraction amount of hypericin reached 4.70 mg/g. The main effect order was extraction time > acetone concentration > extraction temperature > liquid to material ratio. Cytopathic effect(CPE)method was used to assay the anti-infectious bronchitis virus(IBV) effects of the extract of Hypericum attenuatum Choisy. The extract was added before, after and at the same time as the infection of infectious bronchitis virus, the result showed it had certain inhibitory effect on cytopathic effect, indicated that the extract of Hypericum attenuatum Choisy had certain anti-IBV effect.
Based on the single factor experiment, the response surface method was used to optimize the key extraction factors affecting ultrasonic extraction of hypericin from Hypericum attenuatum Choisy with the extraction amount of hypericin as evaluation index and a mathematical model was established. The optimum conditions for ultrasonic extraction of hypericin were as follows: extraction temperature 41 ℃,acetone volume fraction 69.5%, extraction time 43.5 min, liquid to material ratio 15.3. Under these conditions, the extraction amount of hypericin reached 4.70 mg/g. The main effect order was extraction time > acetone concentration > extraction temperature > liquid to material ratio. Cytopathic effect(CPE)method was used to assay the anti-infectious bronchitis virus(IBV) effects of the extract of Hypericum attenuatum Choisy. The extract was added before, after and at the same time as the infection of infectious bronchitis virus, the result showed it had certain inhibitory effect on cytopathic effect, indicated that the extract of Hypericum attenuatum Choisy had certain anti-IBV effect.
引文
[1]Zhai X J,Chen F,Chen C,et al.LC-MS/MS based studies on the anti-depressant effect of hypericin in the chronic unpredictable mild stress rat model[J].J Ethnopharmacology,2015,169:363-369.
[2]Assadi A,Zarrindast M R,Jouyban A,et al.Comparing of the effects of hypericin and synthetic antidepressants on the expression of morphine-induced conditioned place preference[J].Ira J Pharm Res,2011,10(3):619-626.
[3]Qu X B,Su Z M,Hu D H,et al.Studies on molecular structure of hypericin and its interactions with HIV-1 protease by molecular modeling[J].Chem J Chinese U,2009,30(7):1402-1405.
[4]Jia Yongrui(贾永蕊),Hu Ran(胡然),Zhang Yonghe(张永鹤),et al.In vitro study on the effects of hypericin and lysine hydrochloride against herpes virus[J].Chinese Journal of Clinical Rehabilitation(中国临床康复),2004,8(5):996-997.
[5]Wang Shuyang(王曙阳),Liang Jianping(梁剑平),Chen Jihong(陈积红),et al.Studies on the inhibitory effects of hypericin on the adsorption ability of foot-and-mouth virus to host cells in vitro[J].Journal of Traditional Chinese Veterinary Medicine(中兽医医药杂志),2009,28(1):5-8.
[6]Pu Xiuying(蒲秀英),Liang Jianping(梁剑平),Xu Tao(许涛),et al.Study on activity in vitro of hypericin against highly pathogenic PRRSV[J].Chinese Veterinary Science(中国兽医科学),2008,38(9):810-815.
[7]Guo Zhiting(郭志廷),Chen Shien(陈士恩),Liang Jianping(梁剑平),et al.Study on hypericin soluble powder against canine distemper virus in vitro[J].Chinese Veterinary Science(中国兽医科学),2010,40(2):201-204.
[8]Pu X Y,Liang J P,Shang R F,et al.Therapeutic efficacy of Hypericumperforatum L.extract for mice infected with an influenza A virus[J].Can J Physiol Pharm,2012,90(2):123-130.
[9]Shang Ruofeng(尚若锋),Liu Yu(刘宇),Chen Haoran(陈昊然),et al.Therapeutic efficacy of hypericin powder for chickens infected with infectious bursal disease virus[J].Chinese Journal of Veterinary Medicine(中国兽医杂志),2012,48(1):12-15.
[10]Li Y,Zhang J L,Wang B H,et al.Co-delivery of siRNA and hypericin into cancer cells by hyaluronic acid modified PLGA-PEInanoparticles[J].Drug Dev Ind Pharm,2016,42(5):737-746.
[11]Mirmalek S A,Azizi M A,Jangholi E,et al.Cytotoxic and apoptogenic effect of hypericin,the bioactive component of Hypericumperforatum on the MCF-7 human breast cancer cell line[J].Cancer Cell Int,2016,16(3):1-9.
[12]Kilic S A,Selmanoglu G,Yilmaz S,et al.Comparison of phototoxic effects of hypericin-mediated photodynamic therapy in HT-29 and Caco-2 colon cancer cells[J].Turk J Biol,2016,40(6):1202-1218.
[13]Kong Tao(孔涛),Fan Jieping(范杰平),Hu Xiaofang(胡小芳),et al.Optimization of ultrasound-assisted extraction of ursolic acid from Plantagoasiatica L.by response surface methodology[J].Food Science(食品科学),2011,32(6):80-84.
[14]Xiao Weihua(肖卫华),Han Lujia(韩鲁佳),Yang Zengling(杨增玲),et al.Optimization of alcohol extraction techniques of flavonoids from Radix A stragali using response surface methodology[J].Journal of China Agricultural University(中国农业大学学报),2007,12(5):52-56.
[15]Hu Chengxu(胡成旭),Hou Xintong(侯欣彤),Feng Yongning(冯永宁),et al.Study on optimization of extraction conditions of polysaccharide from yunzhi by response surface methodology[J].Science and Technology of Food Industry(食品工业科技),2007,7(7):124-126,130.
[16]Gu Zhongmin(顾忠民),Ma Zhongsen(马忠森),An Jihong(安继红),et al.Determination of NK cytotoxicity by improved MTTassay[J].Journal of Jilin University:Medicine Edition(吉林大学学报:医学版),2003,29(1):119-120.
[17]Guo Kai(郭凯),Liang Jianping(梁剑平),Hua Lanying(华兰英),et al.Optimization of ultrasonic extraction of hypericin in Hypericumperforatum by response surface methodology[J].Chinese Journal of Experimental Traditional Medical Formulae(中国实验方剂学杂志),2011,17(11):7-12.
[18]Lou Qi(娄琦).Study on extraction process of hypericin in Hypericumperforatum L[D].Shenyang:Shenyang Pharmaceutical University(沈阳药科大学),2007,P19.
[19]Anjana A K,Francis P,Gomathy S,et al.In vitro antiviral screening of Psidium Gujava L.against HSV-1 and HSV-2 by CPE inhibition assay[J].Asia J Pharm Clin Res,2014,7(1):102-104.
[20]Zhang Xi(张喜),Sun Guohui(孙国辉),Xu Duoduo(徐多多),et al.Study on extraction and separation of hypericin from Hypericum[J].Jilin Journal of Traditional Chinese Medicine(吉林中医药),2010,30(12):1088-1089.
[2]Assadi A,Zarrindast M R,Jouyban A,et al.Comparing of the effects of hypericin and synthetic antidepressants on the expression of morphine-induced conditioned place preference[J].Ira J Pharm Res,2011,10(3):619-626.
[3]Qu X B,Su Z M,Hu D H,et al.Studies on molecular structure of hypericin and its interactions with HIV-1 protease by molecular modeling[J].Chem J Chinese U,2009,30(7):1402-1405.
[4]Jia Yongrui(贾永蕊),Hu Ran(胡然),Zhang Yonghe(张永鹤),et al.In vitro study on the effects of hypericin and lysine hydrochloride against herpes virus[J].Chinese Journal of Clinical Rehabilitation(中国临床康复),2004,8(5):996-997.
[5]Wang Shuyang(王曙阳),Liang Jianping(梁剑平),Chen Jihong(陈积红),et al.Studies on the inhibitory effects of hypericin on the adsorption ability of foot-and-mouth virus to host cells in vitro[J].Journal of Traditional Chinese Veterinary Medicine(中兽医医药杂志),2009,28(1):5-8.
[6]Pu Xiuying(蒲秀英),Liang Jianping(梁剑平),Xu Tao(许涛),et al.Study on activity in vitro of hypericin against highly pathogenic PRRSV[J].Chinese Veterinary Science(中国兽医科学),2008,38(9):810-815.
[7]Guo Zhiting(郭志廷),Chen Shien(陈士恩),Liang Jianping(梁剑平),et al.Study on hypericin soluble powder against canine distemper virus in vitro[J].Chinese Veterinary Science(中国兽医科学),2010,40(2):201-204.
[8]Pu X Y,Liang J P,Shang R F,et al.Therapeutic efficacy of Hypericumperforatum L.extract for mice infected with an influenza A virus[J].Can J Physiol Pharm,2012,90(2):123-130.
[9]Shang Ruofeng(尚若锋),Liu Yu(刘宇),Chen Haoran(陈昊然),et al.Therapeutic efficacy of hypericin powder for chickens infected with infectious bursal disease virus[J].Chinese Journal of Veterinary Medicine(中国兽医杂志),2012,48(1):12-15.
[10]Li Y,Zhang J L,Wang B H,et al.Co-delivery of siRNA and hypericin into cancer cells by hyaluronic acid modified PLGA-PEInanoparticles[J].Drug Dev Ind Pharm,2016,42(5):737-746.
[11]Mirmalek S A,Azizi M A,Jangholi E,et al.Cytotoxic and apoptogenic effect of hypericin,the bioactive component of Hypericumperforatum on the MCF-7 human breast cancer cell line[J].Cancer Cell Int,2016,16(3):1-9.
[12]Kilic S A,Selmanoglu G,Yilmaz S,et al.Comparison of phototoxic effects of hypericin-mediated photodynamic therapy in HT-29 and Caco-2 colon cancer cells[J].Turk J Biol,2016,40(6):1202-1218.
[13]Kong Tao(孔涛),Fan Jieping(范杰平),Hu Xiaofang(胡小芳),et al.Optimization of ultrasound-assisted extraction of ursolic acid from Plantagoasiatica L.by response surface methodology[J].Food Science(食品科学),2011,32(6):80-84.
[14]Xiao Weihua(肖卫华),Han Lujia(韩鲁佳),Yang Zengling(杨增玲),et al.Optimization of alcohol extraction techniques of flavonoids from Radix A stragali using response surface methodology[J].Journal of China Agricultural University(中国农业大学学报),2007,12(5):52-56.
[15]Hu Chengxu(胡成旭),Hou Xintong(侯欣彤),Feng Yongning(冯永宁),et al.Study on optimization of extraction conditions of polysaccharide from yunzhi by response surface methodology[J].Science and Technology of Food Industry(食品工业科技),2007,7(7):124-126,130.
[16]Gu Zhongmin(顾忠民),Ma Zhongsen(马忠森),An Jihong(安继红),et al.Determination of NK cytotoxicity by improved MTTassay[J].Journal of Jilin University:Medicine Edition(吉林大学学报:医学版),2003,29(1):119-120.
[17]Guo Kai(郭凯),Liang Jianping(梁剑平),Hua Lanying(华兰英),et al.Optimization of ultrasonic extraction of hypericin in Hypericumperforatum by response surface methodology[J].Chinese Journal of Experimental Traditional Medical Formulae(中国实验方剂学杂志),2011,17(11):7-12.
[18]Lou Qi(娄琦).Study on extraction process of hypericin in Hypericumperforatum L[D].Shenyang:Shenyang Pharmaceutical University(沈阳药科大学),2007,P19.
[19]Anjana A K,Francis P,Gomathy S,et al.In vitro antiviral screening of Psidium Gujava L.against HSV-1 and HSV-2 by CPE inhibition assay[J].Asia J Pharm Clin Res,2014,7(1):102-104.
[20]Zhang Xi(张喜),Sun Guohui(孙国辉),Xu Duoduo(徐多多),et al.Study on extraction and separation of hypericin from Hypericum[J].Jilin Journal of Traditional Chinese Medicine(吉林中医药),2010,30(12):1088-1089.
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