胡桃楸茎枝水提物的急性毒性及体外抗肿瘤活性研究
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摘要
目的研究胡桃楸水提物的急性毒性以及体外抗肿瘤活性和毒性,以期评价胡桃楸水提物的毒副作用及对肿瘤细胞的选择性毒性。方法采用热回流的方法制备胡桃楸水提物。体内急性毒性研究以小鼠为模型;体外抗肿瘤活性和毒性分别以胡桃楸水提物对人胃癌细胞(HGC-27)和高分化的人神经母细胞瘤细胞(SH-SY5Y)的增殖抑制作用作为评价指标,采用MTT比色法评价胡桃楸水提物对HGC-27和SH-SY5Y细胞形态和相对增殖率(RGR)的影响,进而评价胡桃楸水提物对肿瘤细胞的选择性抑制作用。结果胡桃楸水提物冻干粉的得率为12.5%。口服给药后,在最大给药剂量下(以生药计为227.27g·kg~(-1)),胡桃楸水提物对小鼠未表现出毒性。在体外抗肿瘤作用与毒性研究中,加入不同浓度胡桃楸水提物,当浓度≤0.1 mg·m L-1时,对两种细胞未显示细胞毒性。当浓度≥0.5 mg·m L~(-1)时,HGC-27细胞的RGR≤51.48%,有明显的细胞毒性;而对SH-SY5Y细胞无细胞毒性。结论胡桃楸水提物对小鼠无明显毒副作用。在低浓度下,胡桃楸水提物对HGC-27细胞和SH-SY5Y细胞均无明显毒性作用;在高浓度下,对HGC-27细胞显示一定的细胞毒性,而对SH-SY5Y细胞仍无明显毒性,说明胡桃楸水提物在高浓度下对胃癌细胞的增殖抑制作用具有一定的选择性。
Objective To determine the side effect of the water extract of Juglans mandshurica, and evaluate the acute toxicity of the extract. The selective anti-tumor activity of the extract for tumor cells, the in vitro anti-tumor activity and toxicity of the extract. Methods The extract was obtained by extracting the branches of Juglans mandshurica with water under reflux. The maximum dose of the extract was orally administered to the mice and the general state and death of the mice were continuously observed for 14 days. The acute toxicity of the extract.To research the selective anti-tumor activity of the extracts, the in vitro anti-tumor activity and toxicity of the extract were respectively evaluated by the inhibitory effects on the growth of human gastric cancer cells(HGC-27) and human neuroblastoma cells(SH-SY5 Y). The morphology of the two cells was observed and the relative growth rate(RGR) was determined by MTT method. Results The yield of the lyophilized water extract of the Juglans mandshurica was 12.5%. After the oral administration, the extract did not show toxicity on mice at the maximum dose of 227.27 g·kg~(-1)(crude drug). After adding the extract at concentrations ≤ 0.1 mg·m L~(-1), the RGR of SH-SY5 Y and HGC-27 cells was both > 100%, showing no cytotoxicity to the tested cells. When the concentration of the extract ≥ 0.5 mg·m L~(-1), the RGR of HGC-27 cells ≤ 51.48%, with obvious toxicity, while implied no toxicity to SH-SY5 Y cells. Conclusion The water extract of Juglans mandshurica has no toxicity on mice. At the low concentrations, the extract does not exhibit cytotoxicity to both SH-SY5 Y and HGC-27 cells.While at the high concentration, the extract shows no cytotoxicity to SH-SY5 Y cells but cytotoxicity to HGC-27 cells, suggesting selective inhibition on the growth of HGC-27 cells.
Objective To determine the side effect of the water extract of Juglans mandshurica, and evaluate the acute toxicity of the extract. The selective anti-tumor activity of the extract for tumor cells, the in vitro anti-tumor activity and toxicity of the extract. Methods The extract was obtained by extracting the branches of Juglans mandshurica with water under reflux. The maximum dose of the extract was orally administered to the mice and the general state and death of the mice were continuously observed for 14 days. The acute toxicity of the extract.To research the selective anti-tumor activity of the extracts, the in vitro anti-tumor activity and toxicity of the extract were respectively evaluated by the inhibitory effects on the growth of human gastric cancer cells(HGC-27) and human neuroblastoma cells(SH-SY5 Y). The morphology of the two cells was observed and the relative growth rate(RGR) was determined by MTT method. Results The yield of the lyophilized water extract of the Juglans mandshurica was 12.5%. After the oral administration, the extract did not show toxicity on mice at the maximum dose of 227.27 g·kg~(-1)(crude drug). After adding the extract at concentrations ≤ 0.1 mg·m L~(-1), the RGR of SH-SY5 Y and HGC-27 cells was both > 100%, showing no cytotoxicity to the tested cells. When the concentration of the extract ≥ 0.5 mg·m L~(-1), the RGR of HGC-27 cells ≤ 51.48%, with obvious toxicity, while implied no toxicity to SH-SY5 Y cells. Conclusion The water extract of Juglans mandshurica has no toxicity on mice. At the low concentrations, the extract does not exhibit cytotoxicity to both SH-SY5 Y and HGC-27 cells.While at the high concentration, the extract shows no cytotoxicity to SH-SY5 Y cells but cytotoxicity to HGC-27 cells, suggesting selective inhibition on the growth of HGC-27 cells.
引文
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[22]刘薇,季宇彬.青龙衣毒性作用及体内抗肿瘤作用的实验研究[J].哈尔滨商业大学学报(自然科学版),2004,(1):4-10.
[23]王光,吴宜艳,雷涛.核桃楸皮粗多糖的急性毒性实验研究[J].实验动物科学,2013,30(1):26-27.
[24]Barathi S,Vardhini RDS,Chitra P,et al. Cytotoxic effect of juglone on human peripheral blood lymphocytes[J].Asian J Pharm Clin Res,2013,6(4):178-186.
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[26]Aynehchi Y,Dehpour AR,Mahmoodian M. Juglone:The echtiotoxic principle of Pterocarya fraxinifolia[J]. Phytochemistry,1973,12(12):3001-3002.
[27]田玉玲,李淑红,刘冰,等.胡桃楸水提物的遗传毒性及抗诱变作用研究[J].癌变·畸变·突变,2011,23(3):216-218.
[2]周媛媛,王栋,牛峰.抗肿瘤中药青龙衣化学成分的研究[J].中草药,2010,41(1):11-14.
[3]沈广志,邹桂华,梁婷,等.核桃楸的化学成分研究进展[J].中国实验方剂学杂志,2015,21(17):219-224.
[4]LeeKS,LiG,KimSH,etal.Cytotoxicdiarylheptanoids from the roots of Juglans mandshurica[J]. J Nat Prod,2002,65(11):1707-1708.
[5]李静,徐康平,邹辉,等.胡桃楸青果皮化学成分研究[J].中南药学,2013,11(1):1-3.
[6]辽宁省中药标准编委会.辽宁省中药标准:第一册[S].沈阳:辽宁科学技术出版社,2009:144-146.
[7]贾敏如,李星炜.中国民族药志药[M].北京:中国医药科技出版社,2005:354.
[8]刘国如,徐康平,申健,等.胡桃楸根化学成分研究[J].中南药学,2009,7(9):644-646.
[9]文姝,刘纪文,包永明,等.胡桃楸提取液对肿瘤细胞增殖的抑制作用[J].中国微生态学杂志,2011,23(8):703-705.
[10]林瑞新,房学东,曹宏,等.核桃楸皮提取物对胃癌细胞增殖的影响[J].中国生物制品学杂志,2010,23(3):294-296.
[11]潘丽艳,郭喜平,李淑红,等.胡桃楸树皮提取物对SMMC-7721、MCF-7和A549肿瘤细胞的抑制作用及其机制[J].吉林大学学报(医学版),2009,35(1):124-127.
[12]张妍.胡桃楸树皮有效成分的提取分离与其体外抗肿瘤活性的研究[D].长春:吉林大学,2008.
[13]王添敏,俞文婕,朱林峰,等.胡桃楸不同部位水提取物的体内抗肿瘤作用[J].西北药学杂志,2017,32(5):624-628.
[14]王添敏,俞文婕,邸学,等.加入胡桃醌与敲除胡桃醌的胡桃楸水提物的抗肿瘤活性[J].西北药学杂志,2017,32(4):480-485.
[15]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,2002:1757-1784.
[16]郭忠聪,文静.雷公藤甲素增加乳腺癌细胞对阿霉素敏感性的实验研究[J].中南药学,2018,16(4):469-472.
[17]高丽丽,庞国勋. 5-氟尿嘧啶类衍生物的设计、合成及其抗肿瘤活性研究[J].中南药学,2018,16(4):484-487.
[18]Malhotra JD,Kaufman RJ. The endoplasmic reticulum and the unfolded protein response[J]. Semin Cell Dev Biol,2007,18(6):716-731.
[19]魏飞亭,董嘉皓,唐怡,等.石菖蒲醇提物对SHSY5Y细胞增殖及抗氧化损伤作用[J].江西中医药,2018,49(8):55-57.
[20]谭龙春,吴昌学,赵亮,等.洛伐他汀对抗AβOs引起的SH-SY5Y细胞毒性作用[J].贵州医科大学学报,2017,42(6):650-654,660.
[21]刘河汝,安秀峰,代晓曼,等.植物甾醇对人神经母细胞瘤SH-SY5Y细胞增殖和凋亡的影响[J].中国食品卫生杂志,2013,25(6):497-500.
[22]刘薇,季宇彬.青龙衣毒性作用及体内抗肿瘤作用的实验研究[J].哈尔滨商业大学学报(自然科学版),2004,(1):4-10.
[23]王光,吴宜艳,雷涛.核桃楸皮粗多糖的急性毒性实验研究[J].实验动物科学,2013,30(1):26-27.
[24]Barathi S,Vardhini RDS,Chitra P,et al. Cytotoxic effect of juglone on human peripheral blood lymphocytes[J].Asian J Pharm Clin Res,2013,6(4):178-186.
[25]Paulsen MT,Ljungman M. The natural toxin juglone causes degradation of p53 and induces rapid H2AX phosphorylation and cell death in human fibroblasts[J]. Toxicol Appl Pharmacol,2005,209(1):1-9.
[26]Aynehchi Y,Dehpour AR,Mahmoodian M. Juglone:The echtiotoxic principle of Pterocarya fraxinifolia[J]. Phytochemistry,1973,12(12):3001-3002.
[27]田玉玲,李淑红,刘冰,等.胡桃楸水提物的遗传毒性及抗诱变作用研究[J].癌变·畸变·突变,2011,23(3):216-218.