1. Department of Pharmaceutical Analysis, Xuzhou Medical University, Xuzhou 221004, China; 2. Key Laboratory for Traditional Chinese Medicine Drug Release System under State Administration of Traditional Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210028, China
Abstract：OBJECTIVE To study the mechanism of different mixture ratio steamed notoginseng and ginseng herb pairs (steamed notoginseng and ginseng (1:1, 1:2, 2:1) in hemolytic anemia rats induced by N-acetyl phenyl hydrazine (APH) by UPLC-Q-TOF/MS technique. METHODS In the experiment, rats were injected subcutaneously with APH to replicate the hemolysis and blood deficiency model, taking normal rats as controls, UPLC-Q-TOF/MS technique was used in combination with principal component analysis (PCA) and partial least squares-discrimination analysis (PLS-DA) to analyze the plasma sample metabolic profiles of each group. RESULTS The results of blood routine and organ index determination demonstrated that the blood deficiency model was successfully replicated. Through analysis of metabolic profiles between the normal group and the model group, 20 potential biomarkers were examined in plasma. Among them, eleven metabolites such as protoporphyrin IX and oleamide were up-regulated, while nine metabolites such as L-Histidine and L-Norleucine were down-regulated. In the experiment, steamed notoginseng, ginseng, different mixture ratio herb pairs (steamed notoginseng and ginseng ) and positive medicine (angelica blood tonic oral liquid) were used to intervene in APH-induced hemolytic anemia model rats. The metabolic level of biomarkers could be changed to a normal state after the rats were given steamed notoginseng, ginseng, different mixture ratio herb pairs (steamed notoginseng and ginseng) and positive medicine, steamed panax notoginseng, ginseng, herb pair 1:1, herb pair 1:2, herb pair 2:1, positive drugs jointly regulated the metabolism of L-leucine and L-isoleucine; among them, steamed panax notoginseng, herb pair 1:1, herb pair 2:1 could adjust the metabolism of retinol and tryptophan, and it was not found that ginseng and herb pair 1:1 regulate these two metabolites; in addition, herb pair 1:1 and drug pair 2:1 could regulate the metabolism of histidine, while the other three groups did not regulate the metabolism of histidine; compared with the 1:1 herb pair, the 2:1 herb pair also regulated the metabolism of arachidonic acid. It was speculated that steamed notoginseng and ginseng herb pair (1:1) may play a role of blood tonifying by improving arachidonic acid metabolism, ether lipid metabolism, tryptophan metabolism, while steamed notoginseng and ginseng herb pair (1:2) may play a role of blood tonifying by improving glycerophospholipid metabolism, and steamed notoginseng and ginseng herb pair (2:1) may play a role of blood tonifying by improving arachidonic acid metabolism, histidine metabolism, ether lipid metabolism, and tryptophan metabolism. This study presented evidence for the therapeutic effects of steamed notoginseng, ginseng, different mixture ratio herb pairs (steamed notoginseng and ginseng) on blood deficiency, compared with other groups, the 2:1 group modulated the most metabolites and metabolic pathways; compared with the herb pair 1:1 group, the herb pair 2:1 group participated in the regulation of the arachidonic acid metabolism pathway, the metabolism of arachidonic acid on lipid protein, hemorheology, vascular elasticity, white blood cell function and platelet activation, which has an important regulatory role. CONCULUSION It is inferred that when the ratio of steamed notoginseng to ginseng is 2:1, it has the best effect on hemolytic blood deficiency.
YANG C. The history and origin of notoginseng. Res Pract Chin Med(现代中药研究与实践), 2015, 29(6): 83-86.
[2]
CHEN B, XU H L, JIA X B. Research progress and thought on processing of Panax notoginseng. Chin Tradit Herb Drugs(中草药), 2013, 44(4): 482-487.
[3]
TANG Y P, SHANG E X, CHEN Y Y, et al. Modern research approaches and strategies for compatibility effects and efficacy components of herbal pairs. Acta Pharm Sin(药学学报), 2019, 54(9): 1564-1573.
[4]
LI F, ZHAO Y, LIN R, et al. Research progress on the effective substances and the mechanism of traditional Chinese medicine compound. Chin Pharm J(中国药学杂志), 2019,54(13): 1037-1044.
[5]
WU M H, ZHANG Y, MA Z G, et al. Historical evolution of Panax notoginseng (sanqi) processing methods. China J Chin Mater Med(中国中药杂志) 2018, 43(24): 4923-4928.
[6]
QIAN H L, PAN Z Q, WANG X M, et al. Study of material base on acetylphenylhydrazine induced blood deficiency syndrome model mice. Chin J Integr Med(中国中西医结合杂志), 2018, 38(12): 1490-1495.
[7]
HUANG B, LI J, WANG X Z, et al. Establishment of hemolytic anemia rat model and its laboratory evaluation. J Chongqing Med Univ(重庆医科大学学报), 2010, 35(5): 703-705.
[8]
CHEN R Q, LIAO C B, GUO Q, et al. Combined systems pharmacology and fecal metabonomics to study the biomarkers and therapeutic mechanism of type 2 diabetic nephropathy treated with astragalus and leech. RSC Adv, 2018, 8(48): 27448-27463.
[9]
GUO Q, LI X J, GUO H R, et al. The urinary metabolomics of astragalus radix and dioscoreae rhizomacomes reveal possible therapeutic mechanisms in the treatment of type 2 diabetes. Acta Pharm Sin (药学学报), 2020, 55(1): 83-90.
[10]
CHEN B, CAI T, JIA X B, et al. Simultaneous determination of ten active ginsenosides in steamed notoginseng by UPLC. China J Chin Mater Med(中国中药杂志), 2014, 39(9): 1614-1619.
[11]
ZHOU X H, LONG L L, LI C M, et al. Raw versus steamed panax notogonseng in pharmacological effects. Glob Tradit Chin Med (环球中医药) 2014,7(6): 420-426.
[12]
YANG X J, DENG Y, WU G X, et al. Metabolomic analysis of enriching blood function of angelica sinensis body and tail using UPLC-Q-TOF/MS. Nat Prod Res Dev(天然产物研究与开发), 2018, 30(2): 176-184
[13]
XIE Y F, LI Y X, LIU M C, et al. Study on toxicity mechanism of aconitum carmichaeli lipid-soluble alkaloids to adjuvant-induced arthritis model rats based on plasma metabolomics. J China Pharm(中国药房), 2019,30(1): 78-83.
[14]
YUN X M, DONG S, HU Q Q, et al. 1H-NMR-based metabolomics approach to investigate the urine samples of collagen-induced arthritis rats and the intervention of tetrandrine. J Pharm Biomed Anal, 2018, 154(1): 302-311.
[15]
DENG Y, ZHANG M, ZHANG Y P. Intervention of Gansu DangGui on the mice models with acetyl phenyl hydrazine-induced hemolytic blood-deficiency. West J Tradit Chin Med(西部中医药), 2015, 28(10): 5-8.
[16]
SUN Q H, FU Y P, LI C, et al. Mechanism of Mahuang Xixin Fuzi decoction on the mice model of kidney-yang deficiency syndrome infected with influenza virus A based on serum metabolomics. China J Tradit Chin Med Pharm(中华中医药杂志), 2018, 33(5): 1752-1757.
[17]
WEI W F, CHU Y T, LIU Y, et al. Serum metabonomics in mice infected with mycoplasma pneumoniae by UPLC-Q-TOF-MS. China J Chin Mater Med (中国中药杂志), 2017, 42(7): 1382-1389.
[18]
KANG Z F, ZHUANG Z Y, WAN S J, et al. Study on pathological changes of retina in rats with blood deficiency. Chin J Basic Med Tradit Chin Med(中国中医基础医学杂志), 2001, 136(3): 43-46.
[19]
ZHANG Z Z, FAN M L, HAO X, et al. Integrative drug efficacy assessment of Danggui and European Danggui using NMR-based metabolomics. J Pharm Biomed Anal, 2016, 32(12):12-20.
[20]
YAMADA T, MATSUZAKI M, TANAKA A. Increase in insulin secretion and decrease in muscle degradation by fat-free milk intake are attenuated by physical exercise. Clin Chim Acta, 2018, 484(8): 21-25.
[21]
TAKASHI I, TETSUYA J, TADASHI M, et al. Effect of exercise therapy combined with branched-chain amino acid supplementation on muscle strengthening in persons with osteoarthritis. Hong Kong Physiot J, 2018, 38(1):23-31.
[22]
XU M J, KITAURA Y, SHINDO D, et al. Branched-chain amino acid (BCAA) supplementation enhances adaptability to exercise training of mice with a muscle-specific defect in the control of BCAA catabolism. Biosci Biotechnol Biochem, 2018, 82(5):144-152.
[23]
LU J L, GUO X L, LIANG Y, et al. IL-33 Improves glucose-lipid metabolism disorders in ob/ob mice. Chin Pharm J(中国药学杂志), 2020, 55(19): 1590-1595.
[24]
HUANG Y J, ZHANG D J, ZHANG Y Y, et al. Role of ultrasound and L-lysine/L-argnine in improving the physical stability of myosin-soybean oil emulsion. Food Hydrocoll, 2021, 111(9):106-367.
[25]
CAO H T, Z H X, Z Q C, et al. Mechanism of treatment effect of Huanglian-Huangqin herb pairs on cerebral ischemia rats based on metabolomic approach. China J Chin Mater Med (中国中药杂志), 2017, 42(11): 2159-2167.
[26]
ZHU B D, HAN Y G, GUO W, et al. Effect of different danggui Buxue Tang combination and compatibility on myelosuppression in mice. Chin Pharm J(中国药学杂志), 2008, 55(18): 1386-1390.
[27]
ZHANG S L, WANG Y P, DU Z X, et al. Lipid metabolomics in serum of hyperuricemic rats induced by fructose based on UPC~2-Q / TOF-MS. China J Chin Mater Med (中国中药杂志), 2016, 41(6): 1135-1139.
[28]
ZHOU Y, QIN D Q, ZHANG W, et al. Integrated LC-MS and GC-MS-based untargeted metabolomics studies of the effect of azadirachtin on bactrocera dorsalis larvae. Sci Rep, 2020, 10(1): 205-213.
[29]
CHANG H, LIU Q, BAI W F, et al. Protective effects of Amygdalus mongolica on rats with renal fibrosis based on serum metabolomics. J Ethnopharmacol, 2020, 257(11): 112-858.
[30]
WANG Y, ZOU L H, HOU L H, et al. Application of metabonomics in the treatment of phlegm-dampness polycystic ovary syndrome based on ultrahigh liquid chromatography-mass spectrometry. Lishizhen Med Mater Med Res(时珍国医国药), 2018, 29(7): 1591-1593.
[31]
LUO N C, DING W, QIAN D W, et al. Characteristics of blood nourishing and blood circulation activating efficacy. Chin J Exp Tradit Med Form(中国实验方剂学杂志), 2013,19(4): 240-246.
[32]
ZHOU Y F, NIU M, FANG J X, et al. Metabonomics study on hepatoprotective effect of schisandrae Chinensis fructus based on UPLC-Q-TOF-MS. China J Chin Mater Med (中国中药杂志), 2018, 43(18): 3756-3763.
[33]
QU T L, WANG E B, LI Z Y, et al. Metabolomics of ethyl acetate extract from Huangqi Injection on leucopenia mice. Chin Tradit Pat Med(中成药), 2017, 39(3): 455-461.
[34]
ZHOU X W, ZHANG M Y, DONG B, et al. Inhibitory effect of panax ginseng on proliferation of lewis lung cancer cells with chemical fingerprints and orthogonal experimental design. Chin Pharm J(中国药学杂志), 2019,54(7):563-570.
[35]
PAYMAN S H B, GERARD S, VISVIKIS S. Influence of inflammation on cardiovascular protective effects of cytochrome P450 epoxygenase-derived epoxyeicosatrienoic acids. Drug Metab Rev, 2014, 46(1):33-56.
[36]
XIE C, LI X, WU J, et al. Anti-inflammatory activity of magnesium isoglycyrrhizinate through inhibition of phospholipase A2/arachidonic acid pathway. J Inflamm, 2015, 10(15): 140-152.
[37]
FERNANDO M R, GIEMBYCZ M A. Bidirectional crosstalk via IL-6, PGE2 and PGD2 between murine myofibroblasts and alternatively activated macrophages enhances anti-inflammatory phenotype in both cells. Br J Pharmacol, 2016, 30(9): 1792-1799.
[38]
LI H M, XIANG J Y, WU J, et al. Research progress of leukotriene B4/leukotriene B4 receptor pathway in the mechanism of cerebral ischemic injury. China Med Her(中国医药导报), 2020,17(31):50-53.
[39]
WANG W T, XUE M, YANG L, et al. Experimental research on the protective effect of panax notoginseng saponins on the gastric mucosa and the anti-platelet effect of aspirin based on arachidonic acid metabolic pathway. Chin J Integr Med Cardia-cerebrovasc Dis(中西医结合心脑血管病杂志), 2019, 17(9): 1315-1320.
2021, Vol. 56 
