4味常见中药对免疫抑制小鼠免疫功能及肠道菌群的影响
|
摘要
试验旨在研究4味中药对免疫抑制小鼠免疫功能和肠道菌群的影响,选取70只6周龄雌性昆明小鼠(18～20g),随机分为7组,分别为空白对照组、模型组、玉屏风组、乌梅组、蒲公英组、党参组、石榴皮组。采用环磷酰胺(cyclophosphamide,Cy)构建免疫抑制病理模型,从喂药的第3天起,2～7组以75mg/kg体重腹腔注射Cy,每隔1天注射1次,共注射4次,注射期间正常给药,模型组灌胃0.9%生理盐水,直至停止注射Cy的第2天,共给药7d。检测4味中药对免疫抑制小鼠体重、脾脏和胸腺指数、血液指标、分泌型免疫球蛋白A(sIgA)、IgG及肠道菌群的影响。结果显示,与模型组相比,党参、乌梅、蒲公英、石榴皮可显著提高免疫抑制小鼠体重(P<0.05),其中乌梅组效果最好;显著提高免疫抑制小鼠免疫器官指数(P<0.05),但不能达到正常水平,其中蒲公英组脾脏指数最高;各中药组可提高白细胞数目(WBC)、显著提高红细胞数目(RBC)(P<0.05),但不能达到正常水平;各中药组可显著提高免疫球蛋白数量(P<0.05),党参组sIgA、IgG显著高于空白对照组(P<0.05),且和玉屏风组差异不显著(P>0.05);各中药组可显著降低大肠杆菌数量(P<0.05),显著增加双歧杆菌和乳酸杆菌数量(P<0.05),提高免疫抑制小鼠肠道菌群的丰富性和多样性,其中党参和蒲公英组小鼠肠道菌群的丰富度和多样性较好。综上说述,乌梅、蒲公英、石榴皮、党参均能够颉颃Cy对小鼠体重、免疫功能及肠道菌群的影响,可用于免疫增强类药物的开发。
The effects of 4kinds of traditional Chinese medicine on immune performance and intestinal flora in immunosuppressed mice were studied.70 6-week-old male Kunming mice were chosen and assigned to 7groups:Blank control group,model group,YPF group,Fructus Mume group,Herba Taraxaci group,Codonopsis Radixgroup and Pericarpium Granati group.A path-ological model of immunosuppression was established by cyclophosphamide(Cy).From the 3rd day of administration,75mg/(kg·BW)Cy was intraperitoneal injected in groups 2to 7,once every another day for 4times.During the injection,the normal administration was continue to perform for 7d,and 0.9%saline was administered in model group until the second day after the Cy injection was stopped.The effects of 4traditional Chinese medicines on body weight,spleen index,thymus index,blood indicator,sIgA,IgG and intestinal flora of immunosuppressed mice were examined.The results showed that compared with model group,the body weight of immunosuppressed mice were significantly increased by Fructus Mume,Herba Taraxaci,Codonopsis Radix and Pericarpium Granati(P<0.05),and the effect of Fructus Mume was the best.The immune organ index of immunosuppressed mice were significantly increased by 4kinds of Chinese medicines(P<0.05),but could not reach the normal level,of which the spleen index of HerbaTarax acigroup was the highest.The WBC were improved and the RBC were increased significantly(P<0.05),but could not reach the normal level.The concentration of immunoglobulin in each Chinese medicine group was increased,the sIgA and IgG in the Codonopsis Radixgroup were significantly higher than that in the blank control group(P<0.05),buts no significant different with YPF group(P>0.05).The number of Escherichia coli was significantly decreased(P<0.05),the number of Bifidobacteriumand Lactobacillus was significantly increased(P<0.05)and the richness and diversity of intestinal flora of immunosuppressed mice were increased,of which the Codonopsis Radix and Herba Taraxaci groups were better.In conclusion,Fructus Mume,Herba Taraxaci,Codonopsis Radixand Pericarpium Granati could antagonize the effects of Cy on body weight,immune function and intestinal flora in mice,and could be used in the development of immune-enhancing drugs.
The effects of 4kinds of traditional Chinese medicine on immune performance and intestinal flora in immunosuppressed mice were studied.70 6-week-old male Kunming mice were chosen and assigned to 7groups:Blank control group,model group,YPF group,Fructus Mume group,Herba Taraxaci group,Codonopsis Radixgroup and Pericarpium Granati group.A path-ological model of immunosuppression was established by cyclophosphamide(Cy).From the 3rd day of administration,75mg/(kg·BW)Cy was intraperitoneal injected in groups 2to 7,once every another day for 4times.During the injection,the normal administration was continue to perform for 7d,and 0.9%saline was administered in model group until the second day after the Cy injection was stopped.The effects of 4traditional Chinese medicines on body weight,spleen index,thymus index,blood indicator,sIgA,IgG and intestinal flora of immunosuppressed mice were examined.The results showed that compared with model group,the body weight of immunosuppressed mice were significantly increased by Fructus Mume,Herba Taraxaci,Codonopsis Radix and Pericarpium Granati(P<0.05),and the effect of Fructus Mume was the best.The immune organ index of immunosuppressed mice were significantly increased by 4kinds of Chinese medicines(P<0.05),but could not reach the normal level,of which the spleen index of HerbaTarax acigroup was the highest.The WBC were improved and the RBC were increased significantly(P<0.05),but could not reach the normal level.The concentration of immunoglobulin in each Chinese medicine group was increased,the sIgA and IgG in the Codonopsis Radixgroup were significantly higher than that in the blank control group(P<0.05),buts no significant different with YPF group(P>0.05).The number of Escherichia coli was significantly decreased(P<0.05),the number of Bifidobacteriumand Lactobacillus was significantly increased(P<0.05)and the richness and diversity of intestinal flora of immunosuppressed mice were increased,of which the Codonopsis Radix and Herba Taraxaci groups were better.In conclusion,Fructus Mume,Herba Taraxaci,Codonopsis Radixand Pericarpium Granati could antagonize the effects of Cy on body weight,immune function and intestinal flora in mice,and could be used in the development of immune-enhancing drugs.
引文
[1]吕其壮,刘娟,林谦,等.猪免疫抑制性疾病病因学研究进展[J].动物医学进展,2017,38(9):96-100.LYU Q Z,LIU J,LIN Q,et al.Progress on etiology of porcine immunosuppressive diseases[J].Progress in Veterinary Medicine,2017,38(9):96-100.(in Chinese)
[2]罗超应,李锦宇,王贵波,等.免疫抑制性疾病防治的复杂性[J].动物医学进展,2015,36(10):110-114.LUO C Y,LI J Y,WANG G B,et al.Complexity of prevention and control of immunosuppressive diseases[J].Progress in Veterinary Medicine,2015,36(10):110-114.(in Chinese)
[3]QUERE P,RIVAS C,ESYER K,et al.Abundance of IFN-alpha and IFN-gamma mRNA in blood of resistant and susceptible chickens infected with Marek’s disease virus(MDV)or vaccinated with turkey herpesvirus;And MDV inhibition of subsequent induction of IFNgene transcription[J].Archives of Virology,2005,150(3):507-519.
[4]邓博文.家禽病毒性免疫抑制病的分析与对策[J].中国畜牧兽医,2007,34(4):104-107.DENG B W.The countermeasure and analysis of immunosuppressive viral disease in poultry[J].China Animal Husbandry&Veterinary Medicine,2007,34(4):104-107.(in Chinese)
[5]MARKOWSKI-GRIMSRUD C J,SCHAT K A.Infection with chicken anaemia virus impairs the generation of pathogen-specific cytotoxic T lymphocytes[J]Immunology,2003,109(2):283-294.
[6]孙淑红,崔治中,孙爱军,等.母源抗体对同源及异源禽网状内皮增生病病毒感染诱发免疫抑制的预防作用[J].畜牧兽医学报,2007,38(5):488-492.SUN S H,CUI Z Z,SUN A J,et al.Comparisons of preventive effects of maternal antibody on immunosuppression induced by homologous and heterologous reticuloendotheliosis viruses[J].Chinese Journal of Animal and Veterinary Sciences,2007,38(5):488-492.(in Chinese)
[7]MARCUS P I,ROJEKJ M,SEKELLICK M J.Interferon induction and/or production and its suppression by influenza A viruses[J].Journal of Virology,2005,79(5):2880-2890.
[8]王军海,马呈瑞.禽用中药免疫增强剂的研究进展[J].水禽世界,2016,1:42-45.WANG J H,MA C R.The latest research progress of poultry medicine immune enhancer[J].Waterfowl World,2016,1:42-45.(in Chinese)
[9]李义,苏运芳,齐雪峰,等.PCR-DGGE技术研究重组人溶菌酶对小鼠肠道菌群结构的影响[J].中国兽医学报,2013,33(9):1345-1351.LI Y,SU Y F,QI X F,et al.Effect of feeding rhLYZon bacterial community and diversity in the mice intestinal tract by PCR-DGGE[J].Chinese Journal of Veterinary Science,2013,33(9):1345-1351.(in Chinese)
[10]崔治中,崔保安.兽医免疫学[M].北京:中国农业出版社,2004.CUI Z Z,CUI B A.Veterinary Immunology[M].Beijing:China Agriculture Press,2004.(in Chinese)
[11]梁春敏,吴敏毓,夏礼斌.玉屏风散对小鼠白细胞活化作用的血清药理学研究[J].皖南医学院学报,2001,20(4):251-253.LIANG C M,WU M Y,XIA L B.Relationship between methylenetetrahydrofolate reductase gene polymorphism and cerebral vascular disease[J].Acta Academiae Medicinae Wannan,2001,20(4):251-253.(in Chinese)
[12]甘露.玉屏风散多糖对小鼠免疫功能的影响[J].免疫学杂志,2013,29(2):182-184.GAN L.The effects of traditional Chinese drug Yupingfeng San on host immune functions in mice[J].Immunological Journal,2013,29(2):182-184.(in Chinese)
[13]REN Z,HE C,FAN Y,et al.Immune-enhancing activity of polysaccharides fromCyrtomiun macrophy[J].International Journal of Biological Macromolecules,2014,70(8):590-595.
[14]ZHANG J,GAO X,PAN Y,et al.Toxicology and immunology of Ganoderma lucidum polysaccharides in Kunming mice and Wistar rats[J].International Journal of Biological Macromolecules,2016,85:302-310.
[15]许爱华,任莉,郑媛媛,等.银杏外种皮多糖对环磷酰胺诱导的免疫抑制小鼠免疫反应的调节作用[J].中国药理学与毒理学杂志,2008,22(1):69-71.XU A H,REN L,ZHENG Y Y.Immunomodulatory effect of Ginkgo biloba exocarp polysaccharides on immunosuppressive mice induced by cyclophosphamide[J].Chinese Journal of Pharmacology and Toxicology,2008,22(1):69-71.(in Chinese)
[16]李海旺.红细胞免疫学新探[M].南京:南京大学出版社,1993.LI H W.A New Approach to Erythrocyte Immunology[M].Nanjing:Nanjing University Press,1993.(in Chinese)
[17]章晋武.决明子多糖的结构及其对仔猪生长、免疫和肠道微生态的影响研究[D].南昌:南昌大学,2005.ZHANG J W.Studies on the structure of polysaccharide from cassiae seeds and it’s influence on the growth,immunity and intestinal microflora of piglets[D].Nanchang:Nanchang University,2005.(in Chinese)
[18]沈思捷,姚旭,尚红梅,等.小刺猴头菌发酵浸膏多糖对肉仔鸡肠道菌群的影响[J].吉林农业大学学报,2013,35(1):83-88.SHEN S J,YAO X,SHANG H M,et al.Effect of polysaccharides from submerged fermentation extract of Hericium caput-medusae on intestinal pH value and microflora in broilers[J].Journal of Jilin Agricultural University,2013,35(1):83-88.(in Chinese)
[19]DORE J,CORTHIER G.The human intestinal microbiota[J]Gastroentérologie Clinique Et Biologique,2010,34(1):7-15.
[20]QIN J J,LI R Q,RAES J,et al.A human gut microbial gene catalogue established by metagenomic sequencing[J].Nature,2010,464(7285):59-65.
[21]EN W,DUAN J,YIN J,et al.Dietary L-glutamine supplementation modulates microbial community and activates innate immunity in the mouse intestine[J].Amino Acids,2014,46(10):2403-2413.
[22]赵秋芳.黑附片对免疫抑制小鼠免疫功能及肠道菌群的影响[D].西安:陕西师范大学,2017.ZHAO Q F.Effect of Heifu tablet on immune function and intestinal flora in immunosuppressive mice[D].Xi’an:Shaanxi Normal University,2017.(in Chinese)
[23]高慧婕,李晓,李春霞,等.蒲公英多糖对免疫抑制小鼠的免疫增强作用及对TNF-α的调节[J].济宁医学院学报,2016,39(6):391-394.GAO H J,LI X,LI C X,et al.Immunoenhancement of Taraxacum polysaccharides on immunological function in immunosuppression mice and the regulation of TNF-α[J].Journal of Jining Medical College,2016,39(6):391-394.(in Chinese)
[24]徐志立,于彩娜,张明波,等.中药对“脾胃运化水液”功能双向调节的分子机制及其研究模式的建立与应用[J].辽宁中医杂志,2018,45(5):944-948.XU Z L,YU C N,ZHANG M B,et al.Molecular mechanism of bidirectional regulation of Chinese herbs on function of spleen and stomach controlling digestion and establishment and application of research model[J].Liaoning Journal of Traditional Chinese Medicine,2018,45(5):944-948.(in Chinese)
[2]罗超应,李锦宇,王贵波,等.免疫抑制性疾病防治的复杂性[J].动物医学进展,2015,36(10):110-114.LUO C Y,LI J Y,WANG G B,et al.Complexity of prevention and control of immunosuppressive diseases[J].Progress in Veterinary Medicine,2015,36(10):110-114.(in Chinese)
[3]QUERE P,RIVAS C,ESYER K,et al.Abundance of IFN-alpha and IFN-gamma mRNA in blood of resistant and susceptible chickens infected with Marek’s disease virus(MDV)or vaccinated with turkey herpesvirus;And MDV inhibition of subsequent induction of IFNgene transcription[J].Archives of Virology,2005,150(3):507-519.
[4]邓博文.家禽病毒性免疫抑制病的分析与对策[J].中国畜牧兽医,2007,34(4):104-107.DENG B W.The countermeasure and analysis of immunosuppressive viral disease in poultry[J].China Animal Husbandry&Veterinary Medicine,2007,34(4):104-107.(in Chinese)
[5]MARKOWSKI-GRIMSRUD C J,SCHAT K A.Infection with chicken anaemia virus impairs the generation of pathogen-specific cytotoxic T lymphocytes[J]Immunology,2003,109(2):283-294.
[6]孙淑红,崔治中,孙爱军,等.母源抗体对同源及异源禽网状内皮增生病病毒感染诱发免疫抑制的预防作用[J].畜牧兽医学报,2007,38(5):488-492.SUN S H,CUI Z Z,SUN A J,et al.Comparisons of preventive effects of maternal antibody on immunosuppression induced by homologous and heterologous reticuloendotheliosis viruses[J].Chinese Journal of Animal and Veterinary Sciences,2007,38(5):488-492.(in Chinese)
[7]MARCUS P I,ROJEKJ M,SEKELLICK M J.Interferon induction and/or production and its suppression by influenza A viruses[J].Journal of Virology,2005,79(5):2880-2890.
[8]王军海,马呈瑞.禽用中药免疫增强剂的研究进展[J].水禽世界,2016,1:42-45.WANG J H,MA C R.The latest research progress of poultry medicine immune enhancer[J].Waterfowl World,2016,1:42-45.(in Chinese)
[9]李义,苏运芳,齐雪峰,等.PCR-DGGE技术研究重组人溶菌酶对小鼠肠道菌群结构的影响[J].中国兽医学报,2013,33(9):1345-1351.LI Y,SU Y F,QI X F,et al.Effect of feeding rhLYZon bacterial community and diversity in the mice intestinal tract by PCR-DGGE[J].Chinese Journal of Veterinary Science,2013,33(9):1345-1351.(in Chinese)
[10]崔治中,崔保安.兽医免疫学[M].北京:中国农业出版社,2004.CUI Z Z,CUI B A.Veterinary Immunology[M].Beijing:China Agriculture Press,2004.(in Chinese)
[11]梁春敏,吴敏毓,夏礼斌.玉屏风散对小鼠白细胞活化作用的血清药理学研究[J].皖南医学院学报,2001,20(4):251-253.LIANG C M,WU M Y,XIA L B.Relationship between methylenetetrahydrofolate reductase gene polymorphism and cerebral vascular disease[J].Acta Academiae Medicinae Wannan,2001,20(4):251-253.(in Chinese)
[12]甘露.玉屏风散多糖对小鼠免疫功能的影响[J].免疫学杂志,2013,29(2):182-184.GAN L.The effects of traditional Chinese drug Yupingfeng San on host immune functions in mice[J].Immunological Journal,2013,29(2):182-184.(in Chinese)
[13]REN Z,HE C,FAN Y,et al.Immune-enhancing activity of polysaccharides fromCyrtomiun macrophy[J].International Journal of Biological Macromolecules,2014,70(8):590-595.
[14]ZHANG J,GAO X,PAN Y,et al.Toxicology and immunology of Ganoderma lucidum polysaccharides in Kunming mice and Wistar rats[J].International Journal of Biological Macromolecules,2016,85:302-310.
[15]许爱华,任莉,郑媛媛,等.银杏外种皮多糖对环磷酰胺诱导的免疫抑制小鼠免疫反应的调节作用[J].中国药理学与毒理学杂志,2008,22(1):69-71.XU A H,REN L,ZHENG Y Y.Immunomodulatory effect of Ginkgo biloba exocarp polysaccharides on immunosuppressive mice induced by cyclophosphamide[J].Chinese Journal of Pharmacology and Toxicology,2008,22(1):69-71.(in Chinese)
[16]李海旺.红细胞免疫学新探[M].南京:南京大学出版社,1993.LI H W.A New Approach to Erythrocyte Immunology[M].Nanjing:Nanjing University Press,1993.(in Chinese)
[17]章晋武.决明子多糖的结构及其对仔猪生长、免疫和肠道微生态的影响研究[D].南昌:南昌大学,2005.ZHANG J W.Studies on the structure of polysaccharide from cassiae seeds and it’s influence on the growth,immunity and intestinal microflora of piglets[D].Nanchang:Nanchang University,2005.(in Chinese)
[18]沈思捷,姚旭,尚红梅,等.小刺猴头菌发酵浸膏多糖对肉仔鸡肠道菌群的影响[J].吉林农业大学学报,2013,35(1):83-88.SHEN S J,YAO X,SHANG H M,et al.Effect of polysaccharides from submerged fermentation extract of Hericium caput-medusae on intestinal pH value and microflora in broilers[J].Journal of Jilin Agricultural University,2013,35(1):83-88.(in Chinese)
[19]DORE J,CORTHIER G.The human intestinal microbiota[J]Gastroentérologie Clinique Et Biologique,2010,34(1):7-15.
[20]QIN J J,LI R Q,RAES J,et al.A human gut microbial gene catalogue established by metagenomic sequencing[J].Nature,2010,464(7285):59-65.
[21]EN W,DUAN J,YIN J,et al.Dietary L-glutamine supplementation modulates microbial community and activates innate immunity in the mouse intestine[J].Amino Acids,2014,46(10):2403-2413.
[22]赵秋芳.黑附片对免疫抑制小鼠免疫功能及肠道菌群的影响[D].西安:陕西师范大学,2017.ZHAO Q F.Effect of Heifu tablet on immune function and intestinal flora in immunosuppressive mice[D].Xi’an:Shaanxi Normal University,2017.(in Chinese)
[23]高慧婕,李晓,李春霞,等.蒲公英多糖对免疫抑制小鼠的免疫增强作用及对TNF-α的调节[J].济宁医学院学报,2016,39(6):391-394.GAO H J,LI X,LI C X,et al.Immunoenhancement of Taraxacum polysaccharides on immunological function in immunosuppression mice and the regulation of TNF-α[J].Journal of Jining Medical College,2016,39(6):391-394.(in Chinese)
[24]徐志立,于彩娜,张明波,等.中药对“脾胃运化水液”功能双向调节的分子机制及其研究模式的建立与应用[J].辽宁中医杂志,2018,45(5):944-948.XU Z L,YU C N,ZHANG M B,et al.Molecular mechanism of bidirectional regulation of Chinese herbs on function of spleen and stomach controlling digestion and establishment and application of research model[J].Liaoning Journal of Traditional Chinese Medicine,2018,45(5):944-948.(in Chinese)