蛴螬多糖组分HDPS-2Ⅱ的理化性质及对肝癌的抑制作用
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摘要
目的探讨蛴螬均一性多糖组分HDPS-2Ⅱ对肝癌的抑制作用。方法采用碱水提取,反复冻融除蛋白等方法获得蛴螬总多糖;用离子交换色谱法和尺寸排阻色谱法分离纯化总多糖获得均一性多糖组分HDPS-2Ⅱ。用高效尺寸排阻色谱法(HPSEC)和考马斯亮蓝法等测定HDPS-2Ⅱ的理化性质。采用MTT、划痕实验和H22肝癌移植瘤小鼠模型测定蛴螬多糖体内外的抗肿瘤活性。结果蛴螬多糖组分HDPS-2Ⅱ的相对分子质量为8.2×103 Da,蛋白质、糖醛酸、硫酸基和氨基含量分别为2.08%,12.08%,0.24%和4.89%。HDPS-2Ⅱ体外可抑制肿瘤细胞的迁移,体内可显著抑制小鼠体内肿瘤的生长,HDPS-2Ⅱ中、高剂量组抑瘤率分别为52.0%和56.9%;与环磷酰胺(CTX)组比较,蛴螬多糖组分HDPS-2Ⅱ可显著增高脾脏指数、白细胞和淋巴细胞数量(P<0.05)。结论蛴螬多糖组分HDPS-2Ⅱ具有抑制细胞体外迁移、小鼠移植瘤的生长和免疫调节作用,具有进一步研发的价值。
Objective To determine the inhibitory effect on hepatocellular carcinoma of polysaccharide HDPS-2Ⅱfrom Holotrichia diomphalia Bates.Methods Total polysaccharides fromHolotrichia diomphalia Bates were obtained by alkali water extraction,repeated freezing and thawing method,etc.Polysaccharide HDPS-2Ⅱ was obtained by ion exchange chromatography and size exclusion chromatography.The physicochemical properties of HDPS-2Ⅱ were determined by high-performance size-exclusion chromatography(HPSEC)and coomassie brilliant blue method,etc.MTT,wound scratch assay and H22 mice models of liver cancer were used to determine the antitumor activity of polysaccharide HDPS-2Ⅱin vivo andin vitro.Results The molecular weight of HDPS-2Ⅱ was 8.2×103 Da,and the contents of protein,uronic acid,sulfate and amino group were 2.08%,12.08%,0.24%and 4.89%,respectively.HDPS-2Ⅱinhibited cell migrationin vitro and inhibited the growth of tumor in mice and the anti-tumor rates of the high and medium-dose HDPS-2Ⅱ administered group were 52.0% and 56.9%,respectively.Compared with cyclophosphamide group,the polysaccharide HDPS-2Ⅱ was significantly increased the index of spleen,WBC and lymphocyte(P<0.05).Conclusion The results demonstrated that HDPS-2Ⅱ has the potential of inhibiting cell migration in vitro,growth of mouse transplanted tumor and immune regulation,and it has further research and development value.
Objective To determine the inhibitory effect on hepatocellular carcinoma of polysaccharide HDPS-2Ⅱfrom Holotrichia diomphalia Bates.Methods Total polysaccharides fromHolotrichia diomphalia Bates were obtained by alkali water extraction,repeated freezing and thawing method,etc.Polysaccharide HDPS-2Ⅱ was obtained by ion exchange chromatography and size exclusion chromatography.The physicochemical properties of HDPS-2Ⅱ were determined by high-performance size-exclusion chromatography(HPSEC)and coomassie brilliant blue method,etc.MTT,wound scratch assay and H22 mice models of liver cancer were used to determine the antitumor activity of polysaccharide HDPS-2Ⅱin vivo andin vitro.Results The molecular weight of HDPS-2Ⅱ was 8.2×103 Da,and the contents of protein,uronic acid,sulfate and amino group were 2.08%,12.08%,0.24%and 4.89%,respectively.HDPS-2Ⅱinhibited cell migrationin vitro and inhibited the growth of tumor in mice and the anti-tumor rates of the high and medium-dose HDPS-2Ⅱ administered group were 52.0% and 56.9%,respectively.Compared with cyclophosphamide group,the polysaccharide HDPS-2Ⅱ was significantly increased the index of spleen,WBC and lymphocyte(P<0.05).Conclusion The results demonstrated that HDPS-2Ⅱ has the potential of inhibiting cell migration in vitro,growth of mouse transplanted tumor and immune regulation,and it has further research and development value.
引文
[1]张海涛.多糖类化合物抗氧化作用及其机制的研究进展[J].天津药学,2017,29(3):60-63.
[2] Zhong D,Wang H,Liu M,et al.Ganoderma lucidum polysaccharide peptide prevents renal ischemia reperfusion injury via counteracting oxidative stress[J].Sci Rep,2015,(5):16910.
[3]曾俐娟,杨晨露.红花多糖对人卵巢上皮癌细胞增殖、转移的影响及机制探讨[J].江西医药,2017,52(9):901-904.
[4] Yang C M,Zhou Y J,Wang R J,et al.Anti-angiogenic effects and mechanisms of polysaccharides fromAntrodia cinnamomea with different molecular weights[J].J Ethnopharmacol,2009,123:407-412.
[5] Jeurink P V,van Esch B C,Rijnierse A,et a1.Mechanisms underlying immune efects of dietary oligosaeeharides[J].Am J Clin Nutr,2013,98(2):572S-577S.
[6]刘亦洋,饶小勇,王鹏龙,等.天然免疫调节活性多糖类成分研究进展[J].江西中医药,2018,49(422):75-80.
[7] Takashi Ohta,Kie Kusano,Atsushi Ido,et al.Silkrose:a novel acidic polysaccharide from the silkmoth that can stimulate the innate immune response[J].Carbohydr Polym,2016,136:995-1001.
[8] Cao H.Polysaccharides from Chinese tea:recent advance on bioactivity and function[J].Int J Biol Macromol,2013,62:76-79.
[9]霍俊成,李艺杰,李萍,等.药用昆虫多糖的研究进展[J].现代生物医学进展,2017,17(29):5788-5792.
[10]陈智,郑学燕,朱荣刚,等.蛴螬化学成分及药理作用研究进展[J].食品与药品,2014,16(1):62-64.
[11]江苏新医学院.中药大词典[M].上海:上海人民出版社,2002:2379.
[12]刘媛媛,曹蔚,张雅,等.蛴螬生物碱提取工艺及体内外抗肿瘤活性研究[J].西北药学杂志,2014,29(2):160-163.
[13]李基俊,孙抒,杨万山,等.蛴螬提取物对小鼠S180肉瘤的抑制作用[J].中华医学写作杂志,2003,10(15):1356-1358.
[14]张璐,张斌,易延逵.毛蜂窝菌多糖提取工艺优化[J].辽宁中医药大学学报,2017,19(11):62-64.
[15]董蕊,马卓群,卫功庆,等.不同产地东北林蛙卵胶膜中蛋白质和水解氨基酸含量差异比较[J].食品工业科技,2017,38(20):312-316,335.
[16]马虎强,刘洋,张红,等.骆驼刺茎枝粗多糖的理化性质及抗氧化活性研究[J].中国药师,2018,21(1):48-52.
[17]丛建波,王长振,李妍,等.褐藻硫酸多糖硫酸基含量测定—硫酸钡比浊法研究[J].解放军药学学报,2003,19(3):181-183.
[18]董陆陆,于敬海,孙学斌,等.泥鳅分泌多糖特征官能团检测及其抗氧化活性研究[J].哈尔滨医科大学学报,2013,47(3):209-211.
[19]顾湘,倪美鑫,冒小平,等.氧化苦参碱衍生物的生物转化制备及其抗肿瘤活性研究[J].西北药学杂志,2017,32(6):711-714.
[20]曹蔚,裴克,张雅,等.蛴螬的药学研究进展[J].陕西中医,2013,34(2):250-251.
[21]张典,高春风,姜凤良,等.多糖的免疫调节作用研究进展[J].中国老年学杂志,2016,36(21):5448-5450.
[2] Zhong D,Wang H,Liu M,et al.Ganoderma lucidum polysaccharide peptide prevents renal ischemia reperfusion injury via counteracting oxidative stress[J].Sci Rep,2015,(5):16910.
[3]曾俐娟,杨晨露.红花多糖对人卵巢上皮癌细胞增殖、转移的影响及机制探讨[J].江西医药,2017,52(9):901-904.
[4] Yang C M,Zhou Y J,Wang R J,et al.Anti-angiogenic effects and mechanisms of polysaccharides fromAntrodia cinnamomea with different molecular weights[J].J Ethnopharmacol,2009,123:407-412.
[5] Jeurink P V,van Esch B C,Rijnierse A,et a1.Mechanisms underlying immune efects of dietary oligosaeeharides[J].Am J Clin Nutr,2013,98(2):572S-577S.
[6]刘亦洋,饶小勇,王鹏龙,等.天然免疫调节活性多糖类成分研究进展[J].江西中医药,2018,49(422):75-80.
[7] Takashi Ohta,Kie Kusano,Atsushi Ido,et al.Silkrose:a novel acidic polysaccharide from the silkmoth that can stimulate the innate immune response[J].Carbohydr Polym,2016,136:995-1001.
[8] Cao H.Polysaccharides from Chinese tea:recent advance on bioactivity and function[J].Int J Biol Macromol,2013,62:76-79.
[9]霍俊成,李艺杰,李萍,等.药用昆虫多糖的研究进展[J].现代生物医学进展,2017,17(29):5788-5792.
[10]陈智,郑学燕,朱荣刚,等.蛴螬化学成分及药理作用研究进展[J].食品与药品,2014,16(1):62-64.
[11]江苏新医学院.中药大词典[M].上海:上海人民出版社,2002:2379.
[12]刘媛媛,曹蔚,张雅,等.蛴螬生物碱提取工艺及体内外抗肿瘤活性研究[J].西北药学杂志,2014,29(2):160-163.
[13]李基俊,孙抒,杨万山,等.蛴螬提取物对小鼠S180肉瘤的抑制作用[J].中华医学写作杂志,2003,10(15):1356-1358.
[14]张璐,张斌,易延逵.毛蜂窝菌多糖提取工艺优化[J].辽宁中医药大学学报,2017,19(11):62-64.
[15]董蕊,马卓群,卫功庆,等.不同产地东北林蛙卵胶膜中蛋白质和水解氨基酸含量差异比较[J].食品工业科技,2017,38(20):312-316,335.
[16]马虎强,刘洋,张红,等.骆驼刺茎枝粗多糖的理化性质及抗氧化活性研究[J].中国药师,2018,21(1):48-52.
[17]丛建波,王长振,李妍,等.褐藻硫酸多糖硫酸基含量测定—硫酸钡比浊法研究[J].解放军药学学报,2003,19(3):181-183.
[18]董陆陆,于敬海,孙学斌,等.泥鳅分泌多糖特征官能团检测及其抗氧化活性研究[J].哈尔滨医科大学学报,2013,47(3):209-211.
[19]顾湘,倪美鑫,冒小平,等.氧化苦参碱衍生物的生物转化制备及其抗肿瘤活性研究[J].西北药学杂志,2017,32(6):711-714.
[20]曹蔚,裴克,张雅,等.蛴螬的药学研究进展[J].陕西中医,2013,34(2):250-251.
[21]张典,高春风,姜凤良,等.多糖的免疫调节作用研究进展[J].中国老年学杂志,2016,36(21):5448-5450.