基于网络药理学的柴胡镇痛作用机制初探
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摘要
目的通过采用网络药理学方法,系统分析柴胡镇痛的主要作用机制及作用靶标,并初步阐明口服和注射两种给药方式下作用靶标差异,筛选可用于其质量评价的生物靶标并进行验证。方法基于数据库和文献检索收集柴胡的化学成分及其可能的作用靶标,以文献报道结合口服生物利用度和类药性作为区分口服制剂和注射用药药效成分的原则,从Gene Cards数据库中检索疼痛相关靶标,利用DIP蛋白质相互作用建立成分靶标与疼痛靶标的关联关系,构建柴胡镇痛的"药物成分-靶标-疾病"网络,通过网络分析筛选关键作用靶标并阐释其可能的机制。结果根据网络分析结果,共筛选出13个柴胡镇痛作用的主要靶标,包括胰蛋白酶原1、凝血酶原、二肽基肽酶4、乙酰胆碱酯酶、环氧化酶2、激素受体、雌激素受体、核受体辅助激活子1、一氧化氮合酶3等,主要影响细胞增殖、精氨酸分解代谢、血小板活化等过程。针对口服和注射两种不同给药方式下,主要作用靶点差异的分析结果表明,胰蛋白酶原1、环氧化酶2以及一氧化氮合酶3均为两种方式的主要作用靶标,可作为其生物评价的重要靶标;同时,注射剂成分对凝血酶原的过多作用可能与其不良反应有关。本研究选择环氧化酶2对柴胡的药效作用进行验证,体外试剂盒检测结果表明,柴胡质量浓度为3. 125～50μg·m L~(-1)时可对环氧化酶2活性产生显著直接抑制作用。结论网络药理学可有效预测不同给药方式下柴胡镇痛的作用靶标及机制,亦可为构建基于镇痛作用的柴胡质量生物评价方法提供参考。
OBJECTIVE To analyze the major analgesic mechanism and target of Bupleuri Radix with a network pharmacology method,clarify the difference targets between oral administration and injection administration,and validate the possible biological target for its quality evaluation. METHODS The chemical components and potential targets of Bupleuri Radix were collected based on the database and literature searching and the analgesic associated targets were searched from Genecards database. All of the targets combined with interactional proteins from DIP database were used to construct a"compound-target-disease"network and the major analgesic mechanism of Bupleuri Radix were then predicted by the key function targets screened from it. RESULTS According to the results of network analysis,13 key targets were screened out,including trypsin-1,prothrombin,dipeptidyl peptidase 4,acetylcholinesterase,COX-2,androgen receptor,estrogen receptor,GABA receptor α1 and NOS-3,which might affect the process of cell proliferation,arginine catabolic process,platelet activation and so on. Trypsinogen-1,cyclooxygenase-2 and nitric oxide synthase-3 were the main targets for both of them,and could be used as the targets for biological evaluation of Bupleuri Radix. At the same time,there were some differences between oral administration and injection of Bupleuri Radix,the excessive effects of injection components on prothrombin may be associated with the adverse reactions of injection. Otherwise,inhibition of COX-2 enzyme activity by in vitro testing showed that Bupleuri Radix has good activity at the concentration of 3. 125-50 μg·m L~(-1). CONCLUSION Network pharmacology provides the potential to predict the analgesic mechanism and to establish new quality evaluation methods of Bupleuri Radix.
OBJECTIVE To analyze the major analgesic mechanism and target of Bupleuri Radix with a network pharmacology method,clarify the difference targets between oral administration and injection administration,and validate the possible biological target for its quality evaluation. METHODS The chemical components and potential targets of Bupleuri Radix were collected based on the database and literature searching and the analgesic associated targets were searched from Genecards database. All of the targets combined with interactional proteins from DIP database were used to construct a"compound-target-disease"network and the major analgesic mechanism of Bupleuri Radix were then predicted by the key function targets screened from it. RESULTS According to the results of network analysis,13 key targets were screened out,including trypsin-1,prothrombin,dipeptidyl peptidase 4,acetylcholinesterase,COX-2,androgen receptor,estrogen receptor,GABA receptor α1 and NOS-3,which might affect the process of cell proliferation,arginine catabolic process,platelet activation and so on. Trypsinogen-1,cyclooxygenase-2 and nitric oxide synthase-3 were the main targets for both of them,and could be used as the targets for biological evaluation of Bupleuri Radix. At the same time,there were some differences between oral administration and injection of Bupleuri Radix,the excessive effects of injection components on prothrombin may be associated with the adverse reactions of injection. Otherwise,inhibition of COX-2 enzyme activity by in vitro testing showed that Bupleuri Radix has good activity at the concentration of 3. 125-50 μg·m L~(-1). CONCLUSION Network pharmacology provides the potential to predict the analgesic mechanism and to establish new quality evaluation methods of Bupleuri Radix.
引文
[1] Ch. P 2015. VolⅠ(中国药典2015年版.一部)[S]. 2015:280.
[2] CHEN C X,LI T L,WANG S R. Pharmacology of Traditional Chinese Medicine(中药药理学)[M]. Shanghai:Shanghai Science and Technology Press,2006:39.
[3] XIE H D,JIA X B,CAI B C,et al. Experimental study on antiinflammatory and analgesic effects of volatileoil of Bupleurum chinense and B. scorzonerifolium[J]. Pharm Clin Res(药学与临床研究),2007,15(2):108-110.
[4] YANG L,WANG X Y,LIU C,et al. Advances in chemical constituents and pharmacological activities of Bupleurum chinense[J]. Inf Tradit Chin Med(中医药信息),2012,29(3):143-144.
[5] WANG S,BAO Y R,MENG X S,et al. Extraction and purification procedure of analgesic effective substances in Bupleurum chinese[J]. Asia Pacific Tradit Med(亚太传统医药),2012,8(11):43-45.
[6] YANG H,YANG L,JIANG L. Study on the anti-inflammatory and analgesic activities of Bupleurum chinense and B. marginatum[J]. J China Pharm(中国药房),2012,23(47):4442-4444.
[7] HU J L,ZHI L,ZHANG L F,et al. Effect of Chaihu Shugan powder on rats with migraine NF-kappa B,C-fos and 5-HT[J].Sci Technol Engineer(科学技术与工程),2016,16(12):205-210.
[8] LIU Y H,CHEN Y S,XIE W,et al. Studies on antiendotoxin action of total saponins from Radix Bupleuri[J]. J Chin Med Mater(中药材),2003,26(6):423-425.
[9] ENGIN H B,GURSOY A,NUSSINOV R,et al. Network-based strategies can help mono-and poly-pharmacology drug discovery:a systems biology view[J]. Curr Pharm Des,2014,20(8):1201-1207.
[10] WANG J,CUI H,WANG R,et al. A systems pharmacology-oriented discovery of a new therapeutic use of the TCM formula Liuweiwuling for liver failure[J]. Sci Reports,2018,8(1):5645.
[11] LI S. Network target:a starting point for traditional Chinese medical network pharmacology[J]. China J Chin Mater Med(中国中药杂志),2011,36(15):2017-2020.
[12] NIU M,ZHANG C E,WU S N,et al. Evaluation of the“multiefficacy of one drug”of traditional Chinese medicines based on the biological target network:towards precision medicine[J].World Sci Technol/Mod Tradit Chin Med Mater Med(世界科学技术-中医药现代化),2017,19(1):44-49.
[13] CHAI X,MENG Y K,BAI Z F,et al. Mechanism of antihepatitis B virus activity of Tonkinensis based on biological targets network[J]. Acta Pharm Sin(药学学报),2018,53(3):396-402.
[14] RU J L,LI P,WANG J,et al. TCMSP:a database of systems pharmacology for drug discovery from herbal medicines[J]. J Cheminform,2014,6:13. doi:10. 1186/1758-2946-6-13.
[15] HUANG L,XIE D,YU Y,et al. TCMID 2. 0:a comprehensive resource for TCM[J]. Nucleic Acids Res,2018,46:1117-1120.
[16] XENARIOS I,RICE D W,SALWINSKI L,et al. DIP:the database of interacting proteins[J]. Nucleic Acids Res,2000,32(1):289-291.
[17] XU X,ZHANG W,HUANG C,et al. A novel chemometric method for the prediction of human oral bioavailability[J]. Int J Mol Sci,2012,13(6):6964-6982.
[18] YU H,CHEN J,XU X,et al. A systematic prediction of multiple drug-target interactions from chemical,genomic,pharmacological data[J]. PLoS One,2012,7(5):e37608.
[19] VERGNOLLE N,BUNNETT N W,SHARKEY K A,et al. Proteinase-activated receptor-2 and hyperalgesia:a novel pain pathway[J]. Nat Med,2 0 0 1,7:8 2 1-8 2 6.
[20] HOOGERWERF W A,SHENOY M,WINSTON J H,et al.Trypsin mediates nociception via theproteinase-activated receptor2:a potentially novel role inpancreatic pain[J]. Gastroenterology,2004,127(1):883-891.
[21] SHI Q,NIE S Q,HUANG L Q. New progression of chemical component and pharmacological studies of Radix Bupleuri[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2002,8(5):53-56.
[22] HUANG W H,ZHANG Y K,HOU X Y,et al. Risk of arthralgia due to dipeptidyl peptidase-4 inhibitors:a Meta-analysis[J].Adv Drug React J(药物不良反应杂志),2016,18(4):261-272.
[23] ZHANG Y,ZHOU X L,SHAO Q,et al. Anti-inflammatory effect of Xiaochaihutang in rats with collagen-induced arthritis and the mechanism[J]. Immunol J(免疫学杂志),2015,31(9):781-785.
[24] CAO F,TANG A M. Effect of Xiaochaihu decoction with different dosages of Bupleurum chinense on the content of MTL,ACHE,NO in gastric antrum in normal rats[J]. Chin J Exp Tradit Med Form(中国实验方剂学杂志),2014,36(3):617-619.
[25] WU R H,ZHANG J S,WU X Y. The pain and control in children hemophilia[J]. Chin J Thromb Hemost(血栓与止血学),2009,15(5):236-240.
[26] WANG Y Z,YIN L J. Effect of local application of hemocoagulase on articular cavity hematocele after arthroscopic surgery[J].Lab Med Clin(检验医学与临床),2015,12(13):1842-1844.
[27] HAN L,WANG X W,SHAN H Y,et al. Research of modified major bupleurum decoctions effect on liver function and blood coagulation mechanism[J]. World Chin Med(世界中医药),2016,11(6):1063-1065.
[28] SZEBENI J. Complement activation-related pseudoallergy:a stress reaction in blood triggered by nanomedicines and biologicals.[J].Mol Immunol,2014,61(2):163-173.
[29] KRISINGER M J,GOEBELER V,LU Z,et al. Thrombin generates previously unidentified C5 products that support the terminal complement activation pathway[J]. Blood,2012,120(8):1717-1725.
[30] LEON L R. Molecular biology of thermoregulation-invited review:cytokine regulation of fever:studies using gene knockout mice[J]. J Appl Physiol,2002,92(6):2648-2655.
[31] NANTEL F. Distribution and regulation of cyclooxygenase-2 in carrageenan-induced inflammation[J]. Br J Pharmacol,1999,128:853-859.
[32] PAN X C,ZHANG X Z. Studies on preparation of nimesulide suppository and its quality control[J]. Chin Pharm J(中国药学杂志),2004,39(3):230-230.
[33] GONG X M,WANG S,CHEN S,et al. Anti-inflammatory action and mechanism on the adjuvant arthritis mouse model of petroleum ether extract from Citrullus lanatus Vine[J]. Chin Pharm J(中国药学杂志),2017,52(19):1680-1684.
[34] HOU H L,HE S X,ZHU Z F,et al. The role of saikosaponind in regulating HIF-1α/COX-2 signal transduction pathway in human hepatocellular carcinoma cells[J]. J Xi'an Jiaotong Univ(Med Sci)(西安交通大学学报:医学版),2011,32(1):80-84.
[35] HE S X,LUO J Y,ZHAO G,et al. Effect of saikosaponins-d on cyciooxygenase-2 expression of human hepatocellular carcinoma cell line SMMC-7721[J]. Chin J Hepatol(中华肝脏病杂志),2006,14(9):712-714.
[36] LUO Y Q,WU K C,SUN A H,et al. Significance of COX-1,COX-2 and i NOS expression in superficial gastritis,gastric mucosal atypical hyper plasia and gastric carcinoma[J]. Chin J Digest(中华消化杂志),2000,20(4):223-226.
[37] KANG J Y,WANG H Y,HUANG B,et al. Effects of weiwei granules on COX-2 and p53 in chronic atrophic gastritis rats[J].Chin J Integr Tradit West Med Digest(中国中西医结合消化杂志),2017,25(8):595-599.
[38] LI S F,ZHOU W,XING Y,et al. Progress on the role of estrogen receptors in pain modulation[J]. Progr Physiol Sci(生理科学进展),2010,41(2):121-124.
[39] LI N,CUI K M,LI W M,et al. Role ofγ-aminobutyric acid and its receptors in pain modulation[J]. Int J Anesthesiol Resuscit(国际麻醉学与复苏杂志),2007,28(4):342-345.
[40] CRAFT R M,MOGIL J S,ALOISI A M. Sex differences in pain and analgesia:the role of gonadal hormones[J]. Eur J Pain,2004,8(5):397-411.
[41] WANG L N,WANG W,JIA T Z. Regulative effect of bupleurum and vinegar-processed bupleurum on the estrogen levels of rats[J]. Acta Chin Med Pharmacol(中医药学报),2014,42(1):56-58.
[42] XIE W,BAO Y,YU L J. Effect of saikosaponin on GABA of hippocampus of chronic kindling induced by pentylenetetrazol in rats[J]. J Xi'an Jiaotong Univ(Med Sci)(西安交通大学学报:医学版),2007,28(1):21-24.
[43] REN H. Acupoint Injection Therapy's Clinical Observation of 456Cases of the Oculomotor Nerve[D]. Wuhan:Hebei Medical University,2012.
[44] FAN D Y. Effects of the chaihu longmu decoction on advanced non-small cell lung cancer[J]. Clin J Chin Med(中医临床研究),2017,9(24):133-135.
[45] DENG C,LI J,TANG X Z. Observation of the short-term effects and long-term recurrence rate of Chaihu Shugansan in the treatment of hyperthyroidism thyroid function[J]. J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2017,19(6):107-110.
[46] ZHU J L. The preventive and therapeutic effect of regulatingthe IL-6/STAT3 signaling pathway with entrapment scattered dachaihu on CP pancreatic fibrosis induced by L-arginine[D].Xi'an:Shaanxi University of Chinese Medicine,2016.
[2] CHEN C X,LI T L,WANG S R. Pharmacology of Traditional Chinese Medicine(中药药理学)[M]. Shanghai:Shanghai Science and Technology Press,2006:39.
[3] XIE H D,JIA X B,CAI B C,et al. Experimental study on antiinflammatory and analgesic effects of volatileoil of Bupleurum chinense and B. scorzonerifolium[J]. Pharm Clin Res(药学与临床研究),2007,15(2):108-110.
[4] YANG L,WANG X Y,LIU C,et al. Advances in chemical constituents and pharmacological activities of Bupleurum chinense[J]. Inf Tradit Chin Med(中医药信息),2012,29(3):143-144.
[5] WANG S,BAO Y R,MENG X S,et al. Extraction and purification procedure of analgesic effective substances in Bupleurum chinese[J]. Asia Pacific Tradit Med(亚太传统医药),2012,8(11):43-45.
[6] YANG H,YANG L,JIANG L. Study on the anti-inflammatory and analgesic activities of Bupleurum chinense and B. marginatum[J]. J China Pharm(中国药房),2012,23(47):4442-4444.
[7] HU J L,ZHI L,ZHANG L F,et al. Effect of Chaihu Shugan powder on rats with migraine NF-kappa B,C-fos and 5-HT[J].Sci Technol Engineer(科学技术与工程),2016,16(12):205-210.
[8] LIU Y H,CHEN Y S,XIE W,et al. Studies on antiendotoxin action of total saponins from Radix Bupleuri[J]. J Chin Med Mater(中药材),2003,26(6):423-425.
[9] ENGIN H B,GURSOY A,NUSSINOV R,et al. Network-based strategies can help mono-and poly-pharmacology drug discovery:a systems biology view[J]. Curr Pharm Des,2014,20(8):1201-1207.
[10] WANG J,CUI H,WANG R,et al. A systems pharmacology-oriented discovery of a new therapeutic use of the TCM formula Liuweiwuling for liver failure[J]. Sci Reports,2018,8(1):5645.
[11] LI S. Network target:a starting point for traditional Chinese medical network pharmacology[J]. China J Chin Mater Med(中国中药杂志),2011,36(15):2017-2020.
[12] NIU M,ZHANG C E,WU S N,et al. Evaluation of the“multiefficacy of one drug”of traditional Chinese medicines based on the biological target network:towards precision medicine[J].World Sci Technol/Mod Tradit Chin Med Mater Med(世界科学技术-中医药现代化),2017,19(1):44-49.
[13] CHAI X,MENG Y K,BAI Z F,et al. Mechanism of antihepatitis B virus activity of Tonkinensis based on biological targets network[J]. Acta Pharm Sin(药学学报),2018,53(3):396-402.
[14] RU J L,LI P,WANG J,et al. TCMSP:a database of systems pharmacology for drug discovery from herbal medicines[J]. J Cheminform,2014,6:13. doi:10. 1186/1758-2946-6-13.
[15] HUANG L,XIE D,YU Y,et al. TCMID 2. 0:a comprehensive resource for TCM[J]. Nucleic Acids Res,2018,46:1117-1120.
[16] XENARIOS I,RICE D W,SALWINSKI L,et al. DIP:the database of interacting proteins[J]. Nucleic Acids Res,2000,32(1):289-291.
[17] XU X,ZHANG W,HUANG C,et al. A novel chemometric method for the prediction of human oral bioavailability[J]. Int J Mol Sci,2012,13(6):6964-6982.
[18] YU H,CHEN J,XU X,et al. A systematic prediction of multiple drug-target interactions from chemical,genomic,pharmacological data[J]. PLoS One,2012,7(5):e37608.
[19] VERGNOLLE N,BUNNETT N W,SHARKEY K A,et al. Proteinase-activated receptor-2 and hyperalgesia:a novel pain pathway[J]. Nat Med,2 0 0 1,7:8 2 1-8 2 6.
[20] HOOGERWERF W A,SHENOY M,WINSTON J H,et al.Trypsin mediates nociception via theproteinase-activated receptor2:a potentially novel role inpancreatic pain[J]. Gastroenterology,2004,127(1):883-891.
[21] SHI Q,NIE S Q,HUANG L Q. New progression of chemical component and pharmacological studies of Radix Bupleuri[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2002,8(5):53-56.
[22] HUANG W H,ZHANG Y K,HOU X Y,et al. Risk of arthralgia due to dipeptidyl peptidase-4 inhibitors:a Meta-analysis[J].Adv Drug React J(药物不良反应杂志),2016,18(4):261-272.
[23] ZHANG Y,ZHOU X L,SHAO Q,et al. Anti-inflammatory effect of Xiaochaihutang in rats with collagen-induced arthritis and the mechanism[J]. Immunol J(免疫学杂志),2015,31(9):781-785.
[24] CAO F,TANG A M. Effect of Xiaochaihu decoction with different dosages of Bupleurum chinense on the content of MTL,ACHE,NO in gastric antrum in normal rats[J]. Chin J Exp Tradit Med Form(中国实验方剂学杂志),2014,36(3):617-619.
[25] WU R H,ZHANG J S,WU X Y. The pain and control in children hemophilia[J]. Chin J Thromb Hemost(血栓与止血学),2009,15(5):236-240.
[26] WANG Y Z,YIN L J. Effect of local application of hemocoagulase on articular cavity hematocele after arthroscopic surgery[J].Lab Med Clin(检验医学与临床),2015,12(13):1842-1844.
[27] HAN L,WANG X W,SHAN H Y,et al. Research of modified major bupleurum decoctions effect on liver function and blood coagulation mechanism[J]. World Chin Med(世界中医药),2016,11(6):1063-1065.
[28] SZEBENI J. Complement activation-related pseudoallergy:a stress reaction in blood triggered by nanomedicines and biologicals.[J].Mol Immunol,2014,61(2):163-173.
[29] KRISINGER M J,GOEBELER V,LU Z,et al. Thrombin generates previously unidentified C5 products that support the terminal complement activation pathway[J]. Blood,2012,120(8):1717-1725.
[30] LEON L R. Molecular biology of thermoregulation-invited review:cytokine regulation of fever:studies using gene knockout mice[J]. J Appl Physiol,2002,92(6):2648-2655.
[31] NANTEL F. Distribution and regulation of cyclooxygenase-2 in carrageenan-induced inflammation[J]. Br J Pharmacol,1999,128:853-859.
[32] PAN X C,ZHANG X Z. Studies on preparation of nimesulide suppository and its quality control[J]. Chin Pharm J(中国药学杂志),2004,39(3):230-230.
[33] GONG X M,WANG S,CHEN S,et al. Anti-inflammatory action and mechanism on the adjuvant arthritis mouse model of petroleum ether extract from Citrullus lanatus Vine[J]. Chin Pharm J(中国药学杂志),2017,52(19):1680-1684.
[34] HOU H L,HE S X,ZHU Z F,et al. The role of saikosaponind in regulating HIF-1α/COX-2 signal transduction pathway in human hepatocellular carcinoma cells[J]. J Xi'an Jiaotong Univ(Med Sci)(西安交通大学学报:医学版),2011,32(1):80-84.
[35] HE S X,LUO J Y,ZHAO G,et al. Effect of saikosaponins-d on cyciooxygenase-2 expression of human hepatocellular carcinoma cell line SMMC-7721[J]. Chin J Hepatol(中华肝脏病杂志),2006,14(9):712-714.
[36] LUO Y Q,WU K C,SUN A H,et al. Significance of COX-1,COX-2 and i NOS expression in superficial gastritis,gastric mucosal atypical hyper plasia and gastric carcinoma[J]. Chin J Digest(中华消化杂志),2000,20(4):223-226.
[37] KANG J Y,WANG H Y,HUANG B,et al. Effects of weiwei granules on COX-2 and p53 in chronic atrophic gastritis rats[J].Chin J Integr Tradit West Med Digest(中国中西医结合消化杂志),2017,25(8):595-599.
[38] LI S F,ZHOU W,XING Y,et al. Progress on the role of estrogen receptors in pain modulation[J]. Progr Physiol Sci(生理科学进展),2010,41(2):121-124.
[39] LI N,CUI K M,LI W M,et al. Role ofγ-aminobutyric acid and its receptors in pain modulation[J]. Int J Anesthesiol Resuscit(国际麻醉学与复苏杂志),2007,28(4):342-345.
[40] CRAFT R M,MOGIL J S,ALOISI A M. Sex differences in pain and analgesia:the role of gonadal hormones[J]. Eur J Pain,2004,8(5):397-411.
[41] WANG L N,WANG W,JIA T Z. Regulative effect of bupleurum and vinegar-processed bupleurum on the estrogen levels of rats[J]. Acta Chin Med Pharmacol(中医药学报),2014,42(1):56-58.
[42] XIE W,BAO Y,YU L J. Effect of saikosaponin on GABA of hippocampus of chronic kindling induced by pentylenetetrazol in rats[J]. J Xi'an Jiaotong Univ(Med Sci)(西安交通大学学报:医学版),2007,28(1):21-24.
[43] REN H. Acupoint Injection Therapy's Clinical Observation of 456Cases of the Oculomotor Nerve[D]. Wuhan:Hebei Medical University,2012.
[44] FAN D Y. Effects of the chaihu longmu decoction on advanced non-small cell lung cancer[J]. Clin J Chin Med(中医临床研究),2017,9(24):133-135.
[45] DENG C,LI J,TANG X Z. Observation of the short-term effects and long-term recurrence rate of Chaihu Shugansan in the treatment of hyperthyroidism thyroid function[J]. J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2017,19(6):107-110.
[46] ZHU J L. The preventive and therapeutic effect of regulatingthe IL-6/STAT3 signaling pathway with entrapment scattered dachaihu on CP pancreatic fibrosis induced by L-arginine[D].Xi'an:Shaanxi University of Chinese Medicine,2016.