甘露消毒丹及其残方体外抗EV7及对EV71感染miR-146a、miR-155、TLR4mRNA表达影响的研究
|
摘要
目的：探讨甘露消毒丹及其残方抗EV71的效应与作用环节,及对EV71所致的炎症及免疫损伤的调节作用,并进一步认识甘露消毒丹结构的科学内涵。
方法：①文献研究：从甘露消毒丹方源、临床运用情况、药理作用等方面探讨甘露消毒丹治“时疫”的有效性。②运用细胞培养技术,设正常细胞对照组、病毒对照组、病毒唑对照组,从直接抑毒实验、治疗性抑毒实验、预防性抑毒实验、预防+治疗性抑毒实验等4组实验分别观察甘露消毒丹全方及清残方、利残方体外抗EV71病毒的细胞存活率、病毒抑制率。③运用实时荧光定量法(RT-qPCR)检测直接抑毒实验中甘露消毒丹全方及清残方、利残方干预后细胞中miRNA-146a、miRNA-155及TLR4的表达变化。
结果：①甘露消毒丹为叶天士创制,首载于《续名医类案》。可广泛运用于临床多种疾病的治疗,其中以内科疾病、传染科疾病为最多,治疗的疾病频次最高的分别是病毒性肝炎、慢性胃炎、手足口病；甘露消毒丹有护肝、抗肝纤维化、解热、抗病毒、抗炎、调节免疫、调节脂质代谢及胃肠功能等作用。甘露消毒丹的临床运用及实验研究均肯定了本方治疗“时疫”的有效性,“疫毒”与“湿热”是其所治“时疫”的重要证候基础。
②在直接抑毒实验、治疗性抑毒实验、预防+治疗性抑毒实验中,甘露消毒丹组及清残方组、利残方组均有比病毒唑组更高的细胞存活率及病毒抑制率；清残方组的细胞存活率及病毒抑制率高于利残方组、甘全方组；在预防性抑毒实验中,甘全方组、清残方组、利残方组、病毒唑组的细胞存活率与病毒对照组接近,病毒抑制率几乎为零。
③甘露消毒丹及其残方均能上调因EV71感染而下降的miRNA-146a、 miRNA-155表达,同时下调因感染而上升的TLR4表达；利残方组的调控作用强于清残方组及甘全方组。
结论：①甘露消毒丹为治湿热性质时疫的良方,值得临床推广,“疫毒”与“湿热”是其所治“时疫”的重要证候基础。
②甘露消毒丹及其残方均有较好的抗手足口病“时邪疫毒”——EV71的作用,并且其抗EV71效应体现在能影响EV71复制的多个环节。
③清热解毒法在方中发挥了更强的抗EV71作用,但利湿化浊法的作用不能忽视,它与清热解毒法相辅相成。
④甘露消毒丹能通过调节miR-146a、miR-155及TLR4来调控EV71所致炎症及免疫反应,提示甘露消毒丹除了能直接抗EV71,还能通过调节免疫与炎症反应来间接抗病毒。值得在治疗手足口病的临床实践中推广
⑤清热解毒法的抗EV71作用强于利湿化浊法,而利湿化浊法对EV71感染所致的炎症及免疫反应的调节作用强于清热解毒法。提示甘露消毒丹中清热解毒药、利湿化浊药各有所长,各司其职,方剂配伍全面合理,不宜偏废。
⑥不论是在直接抗EV71还是在调节炎症及免疫反应中,甘露消毒丹中清热解毒药物及利湿化浊药物之间可能存在相互拮抗的作用,有待进一步研究。
Backgrounds The epidemic situation of hand-foot-mouth disease is getting severer; it is immediacy to find effective ways to treat it. Ganlu Xiaodu Dan is an effective prescription for hand-foot-mouth disease, but the laboratory studies of Ganlu Xiaodu Dan is inadequate, so the mechanism of it still remains unknown.
Objective To explore the anti-EV71effects of Ganlu Xiaodu Dan and its incomplete prescriptions, and explore the regulation and control effects of Ganlu Xiaodu Dan and its incomplete prescriptions in the inflammation and immune injury that induced by EV71,then recognize the scientific connotation of Ganlu Xiaodu Dan's structure further.
Mothods1) Literature research:To discuss Ganlu Xiaodu Dan's effectiveness on epidemic disease from its resource, mechanism of prescription, the situation of its clinical application and its pharmacological action.2) With help of cell culture to observe the anti-EV71effects of Ganlu Xiaodu Dan, and its incomplete prescriptions of Qing prescription and Li prescription, in inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment, preventive-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment.3) To observe changes of expressions of miRNA-146a, miRNA-155and TLR4in RD cells in inhibiting-virus-directly experiment with intervention of Ganlu Xiaodu Dan, Qing prescription and Li prescription by method of RT-qPCR.
Results1) Ganlu Xiaodu Dan was created by Ye Tianshi, first recorded in Xu Ming Yi Lei An. It can be used in treating various clinical diseases, most in internal medicine diseases and infectious diseases. The diseases which be treated most frequently are viral hepatitis, chronic gastritis and hand-foot-and-mouth disease. It can protect liver, anti liver fibrosis, regulate the immune and lipid metabolism and gastrointestinal function, and it has antiviral, anti-inflammatory, antipyretic effects as well. Those clinical uses and experimental studies of Ganlu Xiaodu Dan comfirmed its effectiveness on epidemic disease, the epidemic pathogenic factor and damp and hot is the important syndrome's pathological basis of epidemic disease which Ganlu Xiaodu Dan can treat.2) In inhibiting-virus-directly experiments, therapeutic-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment, groups of Ganlu Xiaodu Dan, Qing prescription and Li prescription had higher cell survival rate and virus inhibition rate than group of ribavirin. Group of Qing prescription had higher cell survival rate and virus inhibition rate than groups of Ganlu Xiaodu Dan and Li prescription. In preventive-inhibiting-virus experiment, cell survival rates of groups of Ganlu Xiaodu Dan, Qing prescription, Li prescription and ribavirin were close to that of the virus control group, and their virus inhibition rates were almost zero.3) Ganlu Xiaodu Dan and its incomplete prescriptions could elevate levels of miRNA-146a and miRNA-155, which dropped when cells infected with EV71. Li prescription's regulatory effect was better than Ganlu Xiaodu Dan and Qing prescription.
Conclusions1) Ganlu Xiaodu Dan is a good prescription for epidemic disease, and it is worthy spreading in clinic. The epidemic pathogenic factor and damp and hot is the important syndrome's basis of epidemic disease that Ganlu Xiaodu Dan can treat.2) Ganlu Xiaodu Dan and its incomplete prescriptions have good antiviral effects of EV71, and they can affect multiple links of EV71's multiplication.3) Heat-clearing and toxin-relieving therapy has stronger anti-EV71effect, but the effect of therapy of removing damp and turbid can not be ignored, they can supplement each other.4) Ganlu Xiaodu Dan can regulate the inflammatory and immune responses which induced by EV71by regulating miR-146a,miR-155and TLR4, which indicates that in addition to direct-anti-EV71effect, Ganlu Xiaodu Dan has an indirect effect of anti-EV71by ways of regulating inflammatory and immune responses. It is worth promoting in treating hand-foot-mouth disease in clinic.5) Heat-clearing and toxin-relieving therapy has stronger anti-EV71effects than therapy of removing damp and turbid, but therapy of removing damp and turbid has stronger effect in regulating inflammatory and immune responses that induced by EV71.This indicates that whether Heat-clearing and toxin-relieving therapy or therapy of removing damp and turbid has its strong points, and each of them can not be deleted at random.6) Whether as for direct-anti-EV71or as for regulating inflammatory and immune responses, herbs of heat-clearing and toxin-relieving therapy and herbs in therapy of removing damp and turbid in Ganlu Xiaodu Dan may have some effects of mutual antagonism, this need to be studied further.
方法：①文献研究：从甘露消毒丹方源、临床运用情况、药理作用等方面探讨甘露消毒丹治“时疫”的有效性。②运用细胞培养技术,设正常细胞对照组、病毒对照组、病毒唑对照组,从直接抑毒实验、治疗性抑毒实验、预防性抑毒实验、预防+治疗性抑毒实验等4组实验分别观察甘露消毒丹全方及清残方、利残方体外抗EV71病毒的细胞存活率、病毒抑制率。③运用实时荧光定量法(RT-qPCR)检测直接抑毒实验中甘露消毒丹全方及清残方、利残方干预后细胞中miRNA-146a、miRNA-155及TLR4的表达变化。
结果：①甘露消毒丹为叶天士创制,首载于《续名医类案》。可广泛运用于临床多种疾病的治疗,其中以内科疾病、传染科疾病为最多,治疗的疾病频次最高的分别是病毒性肝炎、慢性胃炎、手足口病；甘露消毒丹有护肝、抗肝纤维化、解热、抗病毒、抗炎、调节免疫、调节脂质代谢及胃肠功能等作用。甘露消毒丹的临床运用及实验研究均肯定了本方治疗“时疫”的有效性,“疫毒”与“湿热”是其所治“时疫”的重要证候基础。
②在直接抑毒实验、治疗性抑毒实验、预防+治疗性抑毒实验中,甘露消毒丹组及清残方组、利残方组均有比病毒唑组更高的细胞存活率及病毒抑制率；清残方组的细胞存活率及病毒抑制率高于利残方组、甘全方组；在预防性抑毒实验中,甘全方组、清残方组、利残方组、病毒唑组的细胞存活率与病毒对照组接近,病毒抑制率几乎为零。
③甘露消毒丹及其残方均能上调因EV71感染而下降的miRNA-146a、 miRNA-155表达,同时下调因感染而上升的TLR4表达；利残方组的调控作用强于清残方组及甘全方组。
结论：①甘露消毒丹为治湿热性质时疫的良方,值得临床推广,“疫毒”与“湿热”是其所治“时疫”的重要证候基础。
②甘露消毒丹及其残方均有较好的抗手足口病“时邪疫毒”——EV71的作用,并且其抗EV71效应体现在能影响EV71复制的多个环节。
③清热解毒法在方中发挥了更强的抗EV71作用,但利湿化浊法的作用不能忽视,它与清热解毒法相辅相成。
④甘露消毒丹能通过调节miR-146a、miR-155及TLR4来调控EV71所致炎症及免疫反应,提示甘露消毒丹除了能直接抗EV71,还能通过调节免疫与炎症反应来间接抗病毒。值得在治疗手足口病的临床实践中推广
⑤清热解毒法的抗EV71作用强于利湿化浊法,而利湿化浊法对EV71感染所致的炎症及免疫反应的调节作用强于清热解毒法。提示甘露消毒丹中清热解毒药、利湿化浊药各有所长,各司其职,方剂配伍全面合理,不宜偏废。
⑥不论是在直接抗EV71还是在调节炎症及免疫反应中,甘露消毒丹中清热解毒药物及利湿化浊药物之间可能存在相互拮抗的作用,有待进一步研究。
Backgrounds The epidemic situation of hand-foot-mouth disease is getting severer; it is immediacy to find effective ways to treat it. Ganlu Xiaodu Dan is an effective prescription for hand-foot-mouth disease, but the laboratory studies of Ganlu Xiaodu Dan is inadequate, so the mechanism of it still remains unknown.
Objective To explore the anti-EV71effects of Ganlu Xiaodu Dan and its incomplete prescriptions, and explore the regulation and control effects of Ganlu Xiaodu Dan and its incomplete prescriptions in the inflammation and immune injury that induced by EV71,then recognize the scientific connotation of Ganlu Xiaodu Dan's structure further.
Mothods1) Literature research:To discuss Ganlu Xiaodu Dan's effectiveness on epidemic disease from its resource, mechanism of prescription, the situation of its clinical application and its pharmacological action.2) With help of cell culture to observe the anti-EV71effects of Ganlu Xiaodu Dan, and its incomplete prescriptions of Qing prescription and Li prescription, in inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment, preventive-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment.3) To observe changes of expressions of miRNA-146a, miRNA-155and TLR4in RD cells in inhibiting-virus-directly experiment with intervention of Ganlu Xiaodu Dan, Qing prescription and Li prescription by method of RT-qPCR.
Results1) Ganlu Xiaodu Dan was created by Ye Tianshi, first recorded in Xu Ming Yi Lei An. It can be used in treating various clinical diseases, most in internal medicine diseases and infectious diseases. The diseases which be treated most frequently are viral hepatitis, chronic gastritis and hand-foot-and-mouth disease. It can protect liver, anti liver fibrosis, regulate the immune and lipid metabolism and gastrointestinal function, and it has antiviral, anti-inflammatory, antipyretic effects as well. Those clinical uses and experimental studies of Ganlu Xiaodu Dan comfirmed its effectiveness on epidemic disease, the epidemic pathogenic factor and damp and hot is the important syndrome's pathological basis of epidemic disease which Ganlu Xiaodu Dan can treat.2) In inhibiting-virus-directly experiments, therapeutic-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment, groups of Ganlu Xiaodu Dan, Qing prescription and Li prescription had higher cell survival rate and virus inhibition rate than group of ribavirin. Group of Qing prescription had higher cell survival rate and virus inhibition rate than groups of Ganlu Xiaodu Dan and Li prescription. In preventive-inhibiting-virus experiment, cell survival rates of groups of Ganlu Xiaodu Dan, Qing prescription, Li prescription and ribavirin were close to that of the virus control group, and their virus inhibition rates were almost zero.3) Ganlu Xiaodu Dan and its incomplete prescriptions could elevate levels of miRNA-146a and miRNA-155, which dropped when cells infected with EV71. Li prescription's regulatory effect was better than Ganlu Xiaodu Dan and Qing prescription.
Conclusions1) Ganlu Xiaodu Dan is a good prescription for epidemic disease, and it is worthy spreading in clinic. The epidemic pathogenic factor and damp and hot is the important syndrome's basis of epidemic disease that Ganlu Xiaodu Dan can treat.2) Ganlu Xiaodu Dan and its incomplete prescriptions have good antiviral effects of EV71, and they can affect multiple links of EV71's multiplication.3) Heat-clearing and toxin-relieving therapy has stronger anti-EV71effect, but the effect of therapy of removing damp and turbid can not be ignored, they can supplement each other.4) Ganlu Xiaodu Dan can regulate the inflammatory and immune responses which induced by EV71by regulating miR-146a,miR-155and TLR4, which indicates that in addition to direct-anti-EV71effect, Ganlu Xiaodu Dan has an indirect effect of anti-EV71by ways of regulating inflammatory and immune responses. It is worth promoting in treating hand-foot-mouth disease in clinic.5) Heat-clearing and toxin-relieving therapy has stronger anti-EV71effects than therapy of removing damp and turbid, but therapy of removing damp and turbid has stronger effect in regulating inflammatory and immune responses that induced by EV71.This indicates that whether Heat-clearing and toxin-relieving therapy or therapy of removing damp and turbid has its strong points, and each of them can not be deleted at random.6) Whether as for direct-anti-EV71or as for regulating inflammatory and immune responses, herbs of heat-clearing and toxin-relieving therapy and herbs in therapy of removing damp and turbid in Ganlu Xiaodu Dan may have some effects of mutual antagonism, this need to be studied further.
引文
[1]张勇.甘露消毒丹加减治疗慢性乙型肝炎(肝胆湿热证)的临床研究[D].湖北中医学院硕士研究生论文,2009(5)：11
[2]于庆华,于庆民.甘露消毒丹加减治疗急性病毒性肝炎63例[J].内蒙古中医药,2005,(2)：9
[3]农志新.甘露消毒丹加减治疗轻度或中度慢性乙型肝炎72例[J].中医临床研究,2010,2(14)：30-31
[4]叶伟东,许维丹.甘露消毒丹加减治疗病毒性淤胆型肝炎42例[J].现代中西医结合杂志,2003,12(5)：473
[5]褚裕义.甘露消毒丹治疗急性病毒性肝炎临床观察[J].中医杂志,1999,40(20)：87-88
[6]郝明霞,李长秦,王敏,等.加味甘露消毒丹对慢性乙型肝炎血清HBV-DNV影响的临床研究[J].陕西中医,2007,28(1)：14-16
[7]宋诵文.从湿热瘀论治急性黄疸型肝炎38例[J].陕西中医,1994,15(1)：26
[8]夏乃卿.甘露消毒丹治疗小儿急性传染性肝炎26例的疗效观察[J].上海中医药杂志,1999(9)：27
[9]杨晋东.甘露消毒丹加减联合西药治疗慢性乙肝的临床观察[J].内蒙古中医药,2010,(16)：22]
[10]阿力木江 阿不拉.甘露消毒丹加减联合西药治疗慢性乙肝的疗效观察[J].医学信息, 2010,23(5)：154
[11]王利花，李军.手足口病的流行病学及病原学研究进展[J].中国卫生事业管理,2010,264(6)：419-421
[12]田慧,马美美,潘奔前.甘露消毒丹加减治疗手足口病普通病例80例疗效观察[J].新中医,2011,43(6)：76-77
[13]梁志宏,邵振华.甘露消毒丹治疗小儿手足口病120例[J].中国社区医师,2010,12(11)：136
[14]曹军连.甘露消毒丹加减治疗手足口病41例临床观察[J].医学研究杂志,2009,38(3)：106-107
[15]张凡.小儿手足口病的中医辨证施治[J].四川中医,2004,22(1)：15-17
[16]徐华娟.甘露消毒丹加减治疗小儿手足口病48例[J].河南中医,2010,30(9)：876
[17]袁新顺.甘露消毒丹内服加喉症丸外敷治疗带状疱疹[J].现代中西医结合杂志,2003,12(15)：1635
[18]吴敦煌,周虎珍,李凤华.甘露消毒丹内服加中药外敷治疗带状疱疹33例[J].现代中西医结合杂志,2007,16(7)：934
[19]殷振瑾,郭长青,闫远杰.李荣春应用甘露消毒丹经验[J].中医杂志,2008,49(3)：213-214
[20]邵利平.甘露消毒丹杂病应用举隅[J].四川中医,1996.16(8)：52-53
[21]朱会清.甘露消毒丹治疗传染性单核细胞增多症38例[J].实用中医药杂志,2011,27(7)：455
[22]郑邦本,王光富.郑惠伯巧用甘露消毒丹[J].辽宁中医杂志,1992(11)：7
[23]许丰敏.甘露消毒丹加减在流行性出血热发热期中的运用[J].江西中医药,1991,22(1)：20
[241徐建瑞,曾琳.甘露消毒丹的“妙”与“活”[J].贵阳中医学院学报,2007,29(5)：47-48
[25]白跃麻.甘露消毒丹治疗发热疾病的临床运用[J].桂林医学院学报,1992,5(2)：127
[26]袁永耀.甘露消毒丹加减治疗流感150例疗效观察[J].实用医学杂志,1997,13(1)：49-50
[27]姚卫海,周爱国,曲剑华,韩旭.甘露消毒丹加减治疗流感高热的临床分析[J].中西医结合,2003,32(5)：64-65
[28]国家中医药管理局公布中医防治“非典”方案[J].江苏中医药,2003,24(5)：55
[29]李长秦,孙守才,袁晓栋,等.增损甘露消毒丹对肝纤维化大鼠肝组织MMP-1和TIMP-1的影响[J].陕西中医,2010,31(10)：1420-1422
[30]罗媛媛,张晓明,李长秦.增损甘露消毒丹对肝纤维化大鼠血清PCⅢ、 Ⅳ-C影响的实验研究[J].北京中医药大学学报(中医临床版),2009,16(4)：1-3
[31]李长秦,孙守才,袁晓栋,等.增损甘露消毒丹对肝纤维化大鼠血清透明质酸和层勃蛋白的影响[J].吉林中医药,2009,29(6)：540-542
[32]田展飞.甘露消毒丹抗大鼠肝纤维化的实验研究[J].中国中医急症,2011,20(3)：423-424
[33]田展飞.甘露消毒丹对急性肝衰竭大鼠肝细胞凋亡的影响.中国中医急症,2011,20(5)：754,776
[34]杨平.甘露消毒丹对急性肝衰竭大鼠保护作用的实验研究[J].湖北中医药大学学报,2011,13(2)：6-8
[35]程方平,刘松林,杨红兵,等.甘露消毒丹对温病湿热证大鼠LBPmRNA. CD14mRNA、NF-κ Bp动态干预[J].中国实验方剂学杂志,2008,14(4)：56-59
[36]程方平,周洁,陈娟,等.甘露消毒丹对温病湿热证大鼠TLR4mRNA、NF-κBp65表达的影响[J].广州中医药大学学报,2007,24(6)：478-482
[37]程方平,周洁,陈娟,等.甘露消毒丹对湿热证大鼠内毒素转导信号的动态干预[J].中医药学报,2008,36(4)：43-45
[38]程方平,李家庚,刘松林,等.甘露消毒丹对湿热证大鼠模型的干预[J].中医药学报,2008,36(6)：36-38
[39]程方平,刘松林,李云海,等.清热化湿方对温病湿热证大鼠肝线粒体K+2Na+2ATP酶作用的实验研究[J].湖北中医杂志,2007,29(5)：3-5
[40]周璇,程方平.清热化湿方对温病湿热证模型大鼠血清TNF-α IL-1βSIgA的影响[J].辽宁中医药大学学报,2007,9(2)：142-143
[41]徐建瑞,王红梅,曾琳.甘露消毒丹对温病湿热证模型大鼠血清G-CSF、NO的影响[J].贵阳中医学院学报,2009,31(4)：76-77
[42]苏奎国.解毒化湿法治疗湿温病气分证的实验研究[D].山东中医药大学硕士毕业论文,2002
[43]张俊丽,刘妮,周红燕,等.甘露消毒丹加减方体内抗DHBV病毒作用实验研究[J].山西中医学院学报,2008,9(6)：11-13
[44]弓剑.加减甘露消毒丹抗鸭乙型肝炎病毒的实验研究[D].福建中医学院硕士毕业论文, 2009
[45]张志明.清热化湿法干预小鼠病毒性肝炎湿Toll样受体通路的机理研究[D].2010广东中医药大学博士论文：Ⅰ-Ⅲ
[46]贺又舜,伍参荣,赵国荣,等.甘露消毒丹对柯萨奇病毒体外抑制作用的实验研究[J].中国中西医结合杂志,1998,18(12)：737-740
[47]贺又舜,赵国荣,胡建中,等.甘露消毒丹对小鼠IFN、NK及IL-2影响的研究[J].中国实验方剂学杂志,1999：5(3)：9-11
[2]于庆华,于庆民.甘露消毒丹加减治疗急性病毒性肝炎63例[J].内蒙古中医药,2005,(2)：9
[3]农志新.甘露消毒丹加减治疗轻度或中度慢性乙型肝炎72例[J].中医临床研究,2010,2(14)：30-31
[4]叶伟东,许维丹.甘露消毒丹加减治疗病毒性淤胆型肝炎42例[J].现代中西医结合杂志,2003,12(5)：473
[5]褚裕义.甘露消毒丹治疗急性病毒性肝炎临床观察[J].中医杂志,1999,40(20)：87-88
[6]郝明霞,李长秦,王敏,等.加味甘露消毒丹对慢性乙型肝炎血清HBV-DNV影响的临床研究[J].陕西中医,2007,28(1)：14-16
[7]宋诵文.从湿热瘀论治急性黄疸型肝炎38例[J].陕西中医,1994,15(1)：26
[8]夏乃卿.甘露消毒丹治疗小儿急性传染性肝炎26例的疗效观察[J].上海中医药杂志,1999(9)：27
[9]杨晋东.甘露消毒丹加减联合西药治疗慢性乙肝的临床观察[J].内蒙古中医药,2010,(16)：22]
[10]阿力木江 阿不拉.甘露消毒丹加减联合西药治疗慢性乙肝的疗效观察[J].医学信息, 2010,23(5)：154
[11]王利花，李军.手足口病的流行病学及病原学研究进展[J].中国卫生事业管理,2010,264(6)：419-421
[12]田慧,马美美,潘奔前.甘露消毒丹加减治疗手足口病普通病例80例疗效观察[J].新中医,2011,43(6)：76-77
[13]梁志宏,邵振华.甘露消毒丹治疗小儿手足口病120例[J].中国社区医师,2010,12(11)：136
[14]曹军连.甘露消毒丹加减治疗手足口病41例临床观察[J].医学研究杂志,2009,38(3)：106-107
[15]张凡.小儿手足口病的中医辨证施治[J].四川中医,2004,22(1)：15-17
[16]徐华娟.甘露消毒丹加减治疗小儿手足口病48例[J].河南中医,2010,30(9)：876
[17]袁新顺.甘露消毒丹内服加喉症丸外敷治疗带状疱疹[J].现代中西医结合杂志,2003,12(15)：1635
[18]吴敦煌,周虎珍,李凤华.甘露消毒丹内服加中药外敷治疗带状疱疹33例[J].现代中西医结合杂志,2007,16(7)：934
[19]殷振瑾,郭长青,闫远杰.李荣春应用甘露消毒丹经验[J].中医杂志,2008,49(3)：213-214
[20]邵利平.甘露消毒丹杂病应用举隅[J].四川中医,1996.16(8)：52-53
[21]朱会清.甘露消毒丹治疗传染性单核细胞增多症38例[J].实用中医药杂志,2011,27(7)：455
[22]郑邦本,王光富.郑惠伯巧用甘露消毒丹[J].辽宁中医杂志,1992(11)：7
[23]许丰敏.甘露消毒丹加减在流行性出血热发热期中的运用[J].江西中医药,1991,22(1)：20
[241徐建瑞,曾琳.甘露消毒丹的“妙”与“活”[J].贵阳中医学院学报,2007,29(5)：47-48
[25]白跃麻.甘露消毒丹治疗发热疾病的临床运用[J].桂林医学院学报,1992,5(2)：127
[26]袁永耀.甘露消毒丹加减治疗流感150例疗效观察[J].实用医学杂志,1997,13(1)：49-50
[27]姚卫海,周爱国,曲剑华,韩旭.甘露消毒丹加减治疗流感高热的临床分析[J].中西医结合,2003,32(5)：64-65
[28]国家中医药管理局公布中医防治“非典”方案[J].江苏中医药,2003,24(5)：55
[29]李长秦,孙守才,袁晓栋,等.增损甘露消毒丹对肝纤维化大鼠肝组织MMP-1和TIMP-1的影响[J].陕西中医,2010,31(10)：1420-1422
[30]罗媛媛,张晓明,李长秦.增损甘露消毒丹对肝纤维化大鼠血清PCⅢ、 Ⅳ-C影响的实验研究[J].北京中医药大学学报(中医临床版),2009,16(4)：1-3
[31]李长秦,孙守才,袁晓栋,等.增损甘露消毒丹对肝纤维化大鼠血清透明质酸和层勃蛋白的影响[J].吉林中医药,2009,29(6)：540-542
[32]田展飞.甘露消毒丹抗大鼠肝纤维化的实验研究[J].中国中医急症,2011,20(3)：423-424
[33]田展飞.甘露消毒丹对急性肝衰竭大鼠肝细胞凋亡的影响.中国中医急症,2011,20(5)：754,776
[34]杨平.甘露消毒丹对急性肝衰竭大鼠保护作用的实验研究[J].湖北中医药大学学报,2011,13(2)：6-8
[35]程方平,刘松林,杨红兵,等.甘露消毒丹对温病湿热证大鼠LBPmRNA. CD14mRNA、NF-κ Bp动态干预[J].中国实验方剂学杂志,2008,14(4)：56-59
[36]程方平,周洁,陈娟,等.甘露消毒丹对温病湿热证大鼠TLR4mRNA、NF-κBp65表达的影响[J].广州中医药大学学报,2007,24(6)：478-482
[37]程方平,周洁,陈娟,等.甘露消毒丹对湿热证大鼠内毒素转导信号的动态干预[J].中医药学报,2008,36(4)：43-45
[38]程方平,李家庚,刘松林,等.甘露消毒丹对湿热证大鼠模型的干预[J].中医药学报,2008,36(6)：36-38
[39]程方平,刘松林,李云海,等.清热化湿方对温病湿热证大鼠肝线粒体K+2Na+2ATP酶作用的实验研究[J].湖北中医杂志,2007,29(5)：3-5
[40]周璇,程方平.清热化湿方对温病湿热证模型大鼠血清TNF-α IL-1βSIgA的影响[J].辽宁中医药大学学报,2007,9(2)：142-143
[41]徐建瑞,王红梅,曾琳.甘露消毒丹对温病湿热证模型大鼠血清G-CSF、NO的影响[J].贵阳中医学院学报,2009,31(4)：76-77
[42]苏奎国.解毒化湿法治疗湿温病气分证的实验研究[D].山东中医药大学硕士毕业论文,2002
[43]张俊丽,刘妮,周红燕,等.甘露消毒丹加减方体内抗DHBV病毒作用实验研究[J].山西中医学院学报,2008,9(6)：11-13
[44]弓剑.加减甘露消毒丹抗鸭乙型肝炎病毒的实验研究[D].福建中医学院硕士毕业论文, 2009
[45]张志明.清热化湿法干预小鼠病毒性肝炎湿Toll样受体通路的机理研究[D].2010广东中医药大学博士论文：Ⅰ-Ⅲ
[46]贺又舜,伍参荣,赵国荣,等.甘露消毒丹对柯萨奇病毒体外抑制作用的实验研究[J].中国中西医结合杂志,1998,18(12)：737-740
[47]贺又舜,赵国荣,胡建中,等.甘露消毒丹对小鼠IFN、NK及IL-2影响的研究[J].中国实验方剂学杂志,1999：5(3)：9-11