潞党参多糖的抗补体活性分析
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摘要
以山西上党地区著名道地药材潞党参为材料,分析其多糖成分的抗补体活性。首先,通过细胞溶血实验,分析潞党参多糖(Lu Codonopsis pilosula polysaccharides,LCPP)对补体经典途径和旁路途径的影响。结果表明,LCPP具有一定抗补体活性,CH_(50)值为2.061±0.127 mg/mL,AP_(50)值为6.725±0.895 mg/mL,并表现为明显的量效关系。随后,利用人HepG2细胞,通过实时定量PCR实验,分析LCPP对补体系统重要成分C3表达的影响,发现LCPP能够显著抑制肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导的人HepG2细胞中C3的表达。以上结果提示,潞党参是一种潜在的补体抑制剂,在治疗补体过度激活所导致的自身免疫性疾病方面具有应用前景。
The anti-complementary activity of polysaccharides fromLu Codonopsis pilosula(LCPP) was investigated in this paper. The hemolysis assay was used to evaluate the effects of LCPP on the classical and alternative pathways of complement activation. It proved that LCPP repressed the activity of complementary system in a dose-dependent manner, and the CH_(50) and AP_(50) values were 2.061 ±0.127 mg/mL and 6.725 ±0.895 mg/mL, respectively. Furthermore, real-time PCR was used to assess the effect of LCPP on complement component C3 expression in human HepG2 cells. The results showed that LCPP suppressed C3 expression in TNF-α-mediated HepG2 cells significantly. Therefore, Lu Codonopsis pilosula is a potential anti-complementary agent, and may be used in the treatment of autoimmune diseases.
The anti-complementary activity of polysaccharides fromLu Codonopsis pilosula(LCPP) was investigated in this paper. The hemolysis assay was used to evaluate the effects of LCPP on the classical and alternative pathways of complement activation. It proved that LCPP repressed the activity of complementary system in a dose-dependent manner, and the CH_(50) and AP_(50) values were 2.061 ±0.127 mg/mL and 6.725 ±0.895 mg/mL, respectively. Furthermore, real-time PCR was used to assess the effect of LCPP on complement component C3 expression in human HepG2 cells. The results showed that LCPP suppressed C3 expression in TNF-α-mediated HepG2 cells significantly. Therefore, Lu Codonopsis pilosula is a potential anti-complementary agent, and may be used in the treatment of autoimmune diseases.
引文
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[3]Peiris J S M,Chu C M,Cheng V C C,et al.Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia:a prospective study[J].The Lancet,2003,361(9371):1767-1772.
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[6]Namazi S,Sagheb M M,Karimzadeh I.Adverse reactions of immunosuppressive drugs in Iranian adult kidney transplant recipients[J].Experimental and Clinical Transplantation,2012,10(3):224-231.
[7]Hou Y Z,Chen K K,Deng X L,et al.Anti-complementary constituents of Anchusa italica[J].Natural Product Research,2017,31(21):2572-2574.
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[9]王爱青.党参多糖对肾阴虚大鼠抗氧化活性和免疫调节影响[J].中医药临床杂志(Wang Ai-qing.Study on antioxidant and immune regulation effects of Codonopsis polysaccharides on deficiency of kidney-yin type rats[J].Clinical Journal of Traditional Chinese Medicine),2018,30(2):287-290.
[10]李开菊,陈文倩,周霞,等.素花党参多糖对乌鸡生长性能、免疫功能、血常规及肠道菌群的影响[J].四川畜牧兽医(Li Kaiju,Chen Wen-qian,Zhou Xia,et al.Effects of Codonopsis pilosula polysaccharides on the production performance,immune function,routine blood indexes and intestinal microflora of blackbone chicken[J].Sichuan Animal&Veterinary Sciences),2017,10:32-36.
[11]王希春,朱电锋,尹莉莉,等.党参多糖对仔猪生长性能、血清细胞因子及肠黏膜分泌型免疫球蛋白A含量的影响[J].动物营养学报(Wang Xi-chun,Zhu Dian-feng,Yin Li-li,et al.Effects of Codonopsis pilosula polysaccharide on growth performance,serum cytokines and intestinal mucosal secretory immunoglobulin A contents of piglets[J].Chinese Journal of Animal Nutrition),2017,29(11):4069-4075.
[12]刘宽辉,田卫军,高珍珍,等.硒化党参多糖和大蒜多糖协同增强鸡外周血淋巴细胞和新城疫疫苗的免疫功效[J].畜牧兽医学报(Liu Kuan-hui,Tian Wei-jun,Gao Zhen-zhen,et al.Selenizing Codonopsis pilosula polysaccharide and garic polysaccharide synergistically enhance immumological efficacy of chicken lymphocyte and newcastle disease vaccine[J].Acta Veterinaria et Zootechnica Sinica),2017,48(7):1349-1356.
[13]林丹丹,秦韬,任喆,等.硒化党参多糖对免疫抑制小鼠免疫功能的影响[J].中国畜牧兽医(Lin Dan-dan,Qin Tao,Ren Zhe,et al.Effects of selenium-Codonopsis pilosula polysaccharide on immune function of immunosuppressed mice[J].China Animal Husbandry&Veterinary Medicine),2016,43(6):1544-1549.
[14]余兰,王毅.道真洛龙党参多糖对小鼠免疫活性的影响[J].遵义医学院学报(Yu Lan,Wang Yi.Study on immunological activities of polysaccharides isolated from Radix codonopsis on immune function in mice[J].Journal of Zunyi Medical University),2016,39(1):10-13.
[15]胡建燃,郭阳,李平.潞党参多糖的提取及其抗氧化活性分析[J].中国食品添加剂(Hu Jian-ran,Guo Yang,Li Ping.Analysis of Lu Codonopsis pilosula polysaccharide′s extraction and it′s antioxidant activities[J].China Food Additives),2016,7:93-96.
[16]胡建燃,李平,雷海英,等.潞党参多糖对宫颈癌细胞Si Ha增殖和迁移的影响[J].生物技术通报(Hu Jian-ran,Li Ping,Lei Hai-ying,et al.Effects of polysaccharides from Lu Dangshen(Codonopsis pilosula)on proliferation and migration of human cervical Si Ha[J].Biotechnology Bulletin),2017,33(5):159-163.
[17]吴彦,吴甘霖.半枝莲多糖抗补体活性研究[J].中国实验方剂学杂志(Wu Yan,Wu Gan-lin.Studies on anti-complementary activity of the polysacchride from Herba Scutellariae Barbatae[J].Chinese Journal of Experimental Traditional Medical Formulae),2009,15(5):49-52.
[18]Mousavi S,Moradi M,Khorshidahmad T,et al.Anti-inflammatory effects of heparin and its derivatives:a systematic review[J].Advances in Pharmacological Sciences,2015,2015:507151.
[19]Young E.The anti-inflammatory effects of heparin and related compounds[J].Thrombosis Research,2008,122(6):743-752.
[20]阮姝楠,卢燕,陈道峰.广藿香的抗补体活性成分[J].中国中药杂志(Ruan Shu-nan,Lu Yan,Chen Dao-feng.Anti-complementary constituents of Pogostemon cablin[J].China Journal of Chinese Materia Medica),2013,38(13):2129-2135.
[21]吴彦,魏和平,王建波.半枝莲多糖B3-PS2分离纯化及抗补体活性研究[J].药学学报(Wu Yan,Wei He-ping,Wang Jianbo.Anti-complement activity of polysaccharide B3-PS2 puri fied from Herba Scutellariae Barbatae[J].Acta Pharmaceutica Sinica),2009,44(6):615-619.
[22]张娟娟,卢燕,陈道峰.鱼腥草抗补体活性多糖的制备工艺研究[J].中国中药杂志(Zhang Juan-juan,Lu Yan,Chen Daofeng.Preparation procedures of anti-complementary polysaccharides from Houttuynia cordata[J].China Journal of Chinese Materia Medica),2012,37(14):2071-2075.
[23]焦杨,邹录惠,邱莉,等.5种马鞭草科药用植物的抗补体活性[J].中国药科大学学报(Jiao Yang,Zou Lu-hui,Qiu Li,et al.Anticomplementary effects of the extracts of five Verbenaceae herbs in vitro[J].Journal of China Pharmaceutical University),2016,47(4):469-473.
[24]杨涛,郭龙,李灿,等.红芪多糖HPS1-D的化学结构和抗补体活性研究[J].中国中药杂志(Yang Tao,Guo Long,Li Can,et al.Chemical structural features and anti-complementary activity of polysaccharide HPS1-D from Hedysarum polybotrys[J].China Journal of Chinese Materia Medica),2014,39(1):89-93.
[25]姚楠,白杰,张雪梅,等.牛杀菌/通透性增加蛋白对脂多糖介导的炎性细胞因子表达的影响[J].生物工程学报(Yao Nan,Bai Jie,Zhang Xue-mei,et al.Effects on the expression of lipopolysaccharide-induced inflammatory cytokines mediated by bovine bactericidal/permeability-increasing protein[J].Chinese Journal of Biotechnology),2015,31(2):195-205.
[26]洪郁芝,俞东容,朱斌,等.肿瘤坏死因子α诱导人肾近曲小管上皮细胞C3 mRNA表达和蛋白合成及其意义[J].中华肾脏病杂志(Hong Yu-zhi,Yu Dong-rong,Zhu Bin,et al.TNF-αregulation of C3 gene expression and protein biosynthesis in human proximal tubular epithelial cells[J].Chinese Journal of Nephrology),2001,17(5):327-330.
[27]Shavva V S,Mogilenko D A,Dizhe E B,et al.Hepatic nuclear factor 4αpositively regulates complement C3 expression and does not interfere with TNFα-mediated stimulation of C3 expression in Hep G2 cells[J].Gene,2013,524(2):187-192.
[28]褚纯隽,任慧玲,李显伦,等.野马追提取物对脂多糖诱导小鼠急性肺损伤的保护作用[J].华西药学杂志(Chu Chun-jun,Ren Hui-ling,Li Xian-lun,et al.Protective effect of the extracts of Eupatorium lindleyanum on acute lung injury induced by LPS in mice[J].West China Journal of Pharmaceutical Sciences),2015,30(6):653-656.
[29]Steel D M,Whitehead A S.The major acute phase reactants:C-reactive protein,serum amyloid P component and serum amyloid A protein[J].Immunology Today,1994,15(2):81-88.
[30]王志平,韩彦军,刘小宁,等.细菌脂多糖处理对斑马鱼补体基因表达水平的影响[J].水产科学(Wang Zhi-ping,Han Yanjun,Liu Xiao-ning,et al.Effect of bath of lipopolysaccharides derived from bacteria on expression of complement genes in zebrafish[J].Fisheries Science),2014,33(2):115-118.
[2]Emmens R W,Wouters D,Zeerleder S,et al.On the value of therapeutic interventions targeting the complement system in acute myocardial infarction[J].Translational Research,2017,182:103-122.
[3]Peiris J S M,Chu C M,Cheng V C C,et al.Clinical progression and viral load in a community outbreak of coronavirus-associated SARS pneumonia:a prospective study[J].The Lancet,2003,361(9371):1767-1772.
[4]Wong R S,Wu A,To K F,et al.Haematological manifestations in patients with severe acute respiratory syndrome:retrospective analysis[J].British Medical Journal,2003,326(7403):1358-1362.
[5]Barle E L,Winkler G C,Ulrich P,et al.Cancer risk of immunosuppressants in manufacturing[J].Regulatory Toxicology and Pharmacology,2014,70(1):122-124.
[6]Namazi S,Sagheb M M,Karimzadeh I.Adverse reactions of immunosuppressive drugs in Iranian adult kidney transplant recipients[J].Experimental and Clinical Transplantation,2012,10(3):224-231.
[7]Hou Y Z,Chen K K,Deng X L,et al.Anti-complementary constituents of Anchusa italica[J].Natural Product Research,2017,31(21):2572-2574.
[8]赵江燕,王东,高建平.道地药材“潞党参”与常用商品党参鉴别研究[J].中华中医药学刊(Zhao Jiang-yan,Wang Dong,Gao Jian-ping.Pharmacognostic identification of geoherbs Lu Dangshen with commodity codonopsis radix[J].Chinese Archives of Traditional Chinese Medicine),2015,33(1):76-79.
[9]王爱青.党参多糖对肾阴虚大鼠抗氧化活性和免疫调节影响[J].中医药临床杂志(Wang Ai-qing.Study on antioxidant and immune regulation effects of Codonopsis polysaccharides on deficiency of kidney-yin type rats[J].Clinical Journal of Traditional Chinese Medicine),2018,30(2):287-290.
[10]李开菊,陈文倩,周霞,等.素花党参多糖对乌鸡生长性能、免疫功能、血常规及肠道菌群的影响[J].四川畜牧兽医(Li Kaiju,Chen Wen-qian,Zhou Xia,et al.Effects of Codonopsis pilosula polysaccharides on the production performance,immune function,routine blood indexes and intestinal microflora of blackbone chicken[J].Sichuan Animal&Veterinary Sciences),2017,10:32-36.
[11]王希春,朱电锋,尹莉莉,等.党参多糖对仔猪生长性能、血清细胞因子及肠黏膜分泌型免疫球蛋白A含量的影响[J].动物营养学报(Wang Xi-chun,Zhu Dian-feng,Yin Li-li,et al.Effects of Codonopsis pilosula polysaccharide on growth performance,serum cytokines and intestinal mucosal secretory immunoglobulin A contents of piglets[J].Chinese Journal of Animal Nutrition),2017,29(11):4069-4075.
[12]刘宽辉,田卫军,高珍珍,等.硒化党参多糖和大蒜多糖协同增强鸡外周血淋巴细胞和新城疫疫苗的免疫功效[J].畜牧兽医学报(Liu Kuan-hui,Tian Wei-jun,Gao Zhen-zhen,et al.Selenizing Codonopsis pilosula polysaccharide and garic polysaccharide synergistically enhance immumological efficacy of chicken lymphocyte and newcastle disease vaccine[J].Acta Veterinaria et Zootechnica Sinica),2017,48(7):1349-1356.
[13]林丹丹,秦韬,任喆,等.硒化党参多糖对免疫抑制小鼠免疫功能的影响[J].中国畜牧兽医(Lin Dan-dan,Qin Tao,Ren Zhe,et al.Effects of selenium-Codonopsis pilosula polysaccharide on immune function of immunosuppressed mice[J].China Animal Husbandry&Veterinary Medicine),2016,43(6):1544-1549.
[14]余兰,王毅.道真洛龙党参多糖对小鼠免疫活性的影响[J].遵义医学院学报(Yu Lan,Wang Yi.Study on immunological activities of polysaccharides isolated from Radix codonopsis on immune function in mice[J].Journal of Zunyi Medical University),2016,39(1):10-13.
[15]胡建燃,郭阳,李平.潞党参多糖的提取及其抗氧化活性分析[J].中国食品添加剂(Hu Jian-ran,Guo Yang,Li Ping.Analysis of Lu Codonopsis pilosula polysaccharide′s extraction and it′s antioxidant activities[J].China Food Additives),2016,7:93-96.
[16]胡建燃,李平,雷海英,等.潞党参多糖对宫颈癌细胞Si Ha增殖和迁移的影响[J].生物技术通报(Hu Jian-ran,Li Ping,Lei Hai-ying,et al.Effects of polysaccharides from Lu Dangshen(Codonopsis pilosula)on proliferation and migration of human cervical Si Ha[J].Biotechnology Bulletin),2017,33(5):159-163.
[17]吴彦,吴甘霖.半枝莲多糖抗补体活性研究[J].中国实验方剂学杂志(Wu Yan,Wu Gan-lin.Studies on anti-complementary activity of the polysacchride from Herba Scutellariae Barbatae[J].Chinese Journal of Experimental Traditional Medical Formulae),2009,15(5):49-52.
[18]Mousavi S,Moradi M,Khorshidahmad T,et al.Anti-inflammatory effects of heparin and its derivatives:a systematic review[J].Advances in Pharmacological Sciences,2015,2015:507151.
[19]Young E.The anti-inflammatory effects of heparin and related compounds[J].Thrombosis Research,2008,122(6):743-752.
[20]阮姝楠,卢燕,陈道峰.广藿香的抗补体活性成分[J].中国中药杂志(Ruan Shu-nan,Lu Yan,Chen Dao-feng.Anti-complementary constituents of Pogostemon cablin[J].China Journal of Chinese Materia Medica),2013,38(13):2129-2135.
[21]吴彦,魏和平,王建波.半枝莲多糖B3-PS2分离纯化及抗补体活性研究[J].药学学报(Wu Yan,Wei He-ping,Wang Jianbo.Anti-complement activity of polysaccharide B3-PS2 puri fied from Herba Scutellariae Barbatae[J].Acta Pharmaceutica Sinica),2009,44(6):615-619.
[22]张娟娟,卢燕,陈道峰.鱼腥草抗补体活性多糖的制备工艺研究[J].中国中药杂志(Zhang Juan-juan,Lu Yan,Chen Daofeng.Preparation procedures of anti-complementary polysaccharides from Houttuynia cordata[J].China Journal of Chinese Materia Medica),2012,37(14):2071-2075.
[23]焦杨,邹录惠,邱莉,等.5种马鞭草科药用植物的抗补体活性[J].中国药科大学学报(Jiao Yang,Zou Lu-hui,Qiu Li,et al.Anticomplementary effects of the extracts of five Verbenaceae herbs in vitro[J].Journal of China Pharmaceutical University),2016,47(4):469-473.
[24]杨涛,郭龙,李灿,等.红芪多糖HPS1-D的化学结构和抗补体活性研究[J].中国中药杂志(Yang Tao,Guo Long,Li Can,et al.Chemical structural features and anti-complementary activity of polysaccharide HPS1-D from Hedysarum polybotrys[J].China Journal of Chinese Materia Medica),2014,39(1):89-93.
[25]姚楠,白杰,张雪梅,等.牛杀菌/通透性增加蛋白对脂多糖介导的炎性细胞因子表达的影响[J].生物工程学报(Yao Nan,Bai Jie,Zhang Xue-mei,et al.Effects on the expression of lipopolysaccharide-induced inflammatory cytokines mediated by bovine bactericidal/permeability-increasing protein[J].Chinese Journal of Biotechnology),2015,31(2):195-205.
[26]洪郁芝,俞东容,朱斌,等.肿瘤坏死因子α诱导人肾近曲小管上皮细胞C3 mRNA表达和蛋白合成及其意义[J].中华肾脏病杂志(Hong Yu-zhi,Yu Dong-rong,Zhu Bin,et al.TNF-αregulation of C3 gene expression and protein biosynthesis in human proximal tubular epithelial cells[J].Chinese Journal of Nephrology),2001,17(5):327-330.
[27]Shavva V S,Mogilenko D A,Dizhe E B,et al.Hepatic nuclear factor 4αpositively regulates complement C3 expression and does not interfere with TNFα-mediated stimulation of C3 expression in Hep G2 cells[J].Gene,2013,524(2):187-192.
[28]褚纯隽,任慧玲,李显伦,等.野马追提取物对脂多糖诱导小鼠急性肺损伤的保护作用[J].华西药学杂志(Chu Chun-jun,Ren Hui-ling,Li Xian-lun,et al.Protective effect of the extracts of Eupatorium lindleyanum on acute lung injury induced by LPS in mice[J].West China Journal of Pharmaceutical Sciences),2015,30(6):653-656.
[29]Steel D M,Whitehead A S.The major acute phase reactants:C-reactive protein,serum amyloid P component and serum amyloid A protein[J].Immunology Today,1994,15(2):81-88.
[30]王志平,韩彦军,刘小宁,等.细菌脂多糖处理对斑马鱼补体基因表达水平的影响[J].水产科学(Wang Zhi-ping,Han Yanjun,Liu Xiao-ning,et al.Effect of bath of lipopolysaccharides derived from bacteria on expression of complement genes in zebrafish[J].Fisheries Science),2014,33(2):115-118.