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Danggui Buxue Tang (A Chinese Angelica Decoction): A Sample Trial in

Danggui Buxue Tang (A Chinese Angelica Decoction): A Sample Trial in Traditional Chinese Medicine Standardization. Danggui Buxue Tang (A Chinese Angelica Decoction): A Sample Trial in Traditional Chinese Medicine Standardization. 当归补血汤 : 中药黄芪与当归的复方研究. 当归补血汤 : 中药黄芪与当归的复方研究.

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Danggui Buxue Tang (A Chinese Angelica Decoction): A Sample Trial in

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  1. Danggui Buxue Tang (A Chinese Angelica Decoction): A Sample Trial in Traditional Chinese Medicine Standardization Danggui Buxue Tang (A Chinese Angelica Decoction): A Sample Trial in Traditional Chinese Medicine Standardization 当归补血汤:中药黄芪与当归的复方研究 当归补血汤:中药黄芪与当归的复方研究 香港科技大学生物系暨中药研发中心 香港科技大学生物系暨中药研发中心 Supported by by grants from UGC, Area of Excellent (AoE/B-10/01) and RGC (HKUST 6419/06M) of HKSAR

  2. Chinese Medicinal Theory (中医理论) (Sub-healthy Status) Deficiency (Enhance Physiological Function) Supplementation Yang (阳) Qi (气) Blood (血) Yin (阴) Genetic Factors Acquired, Environmental Danggui Buxue Tang (a TCM Formulation) Circulatory System Metabolism Nutrients Radix Astragali 黄芪10 qian (~30 g) Radix Angelicae Sinensis 当归2 qian (~6 g) Neuro-, Endocrino-modulation Bioenergy Production Blood function Immune function 君 Jun (king) 臣 Chen (minister) 佐 Zuo (assistant) 使 Shi (servant) Immuno- modulation Antioxidant Hormone-like Action

  3. Chinese Angelica Decoction Danggui Buxue Tang当归补血汤: Origin and medication (方源与方解) Radix Angelicae Sinensis (Danggui) 当归 Danggui Buxue Tang (DBT;当归补血汤) is the simplest herbal preparation that has been commonly used in Chinese population for about 800 years. The use of DBT was first recorded in Neiwaishang Bianhuo Lun《内外伤辨惑论·暑伤胃气论》by Li Dongyuan (李东垣) in A.D.1247. The ratio of Radix Angelicae sinensis (Danggui,当归) : Radix Astragali (Huangqi, 黄芪) is 1:5. Radix Astragali (Huangqi) 黃芪 Action: To enrich blood, activate blood circulation, regulate menstruation and relieve pain. (具有补气生血的功效) Indications: Anemia with dizziness and palpitation;menstrual disorders amenorrhea, dysmenorrhea; constipation; rheumatic arthragia; traumatic injuries; carbuncles and sores. Dong et al. (2006). J. Agric. Food Chem. 54: 2767-2774 Gao et al. (2006). Planta Medica 72: 1227-1231 Mak et al. (2006). Phytotherapy Research 20 (7): 561-567 Tsim (2005). APBN 8:1316-1321 Tsim et al. (2006). Chinese Medicine Journal (in press) Song et al. (2004). Planta Medica 70: 1222-1227 Gao et al. (2006). FEBS Letter (in press)

  4. Heilongjiang 黑龙江 Beijing 北京 Xinjiang 新疆 Neimenggu 内蒙古 Hebei 河北 Shanxi 山西 Ningxi 宁夏 Sichuan 四川 Hebei 河北 Heilongjian 黑龙江 Shanxi 山西 Neimenggu 內蒙古 10 200 100 5 0 0 Dong et al. (2003). J. Agric. Food Chem. 51:6709-6714 Ma et al. (2000). Natural Medicine 54(5): 213-218 Radix Astragali‚ Huangqi(黃芪) Dried roots of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao (蒙古黄芪) or Astragalus membranaceus (Fisch.) Bge. (膜荚黄芪) Family: Leguminosae. Triterpene saponins (皂苷) and isoflavones, isoflavan and pterocarpans (异黄酮) are considered as active ingredients. Isoflavonoid &saponin Total polysaccharide Total saponin 总皂苷 Total isoflavonoid 总黃酮 Total polysaccharide 总多糖 Ma et al. (2002). J. Agric. Food Chem. 50:4861-4866 Ma et al. (2000). Phytochemistry 54:363-368

  5. Ferulic acid 阿魏酸 Ligustilide 藁本内酯 120 1200 90 900 Content ( mg / 100 g ) 60 Angelica sinensis 当归 Content ( mg / 100 g ) 600 30 300 0 0 Gansu 甘肅 Yunnan 云南 Sichuan 四川 Shanxi 陜西 Japan 日本 Korea 韩国 Gansu 甘肅 Yunnan 云南 Sichuan 四川 Shanxi 陜西 Japan 日本 Korea 韩国 Radix Angelica Sinensis, Danggui (当归) Selection of Angelica roots 和当归 A. acutiloba cultivated in Toyama and Hokkaido of Japan 朝鲜当归 A. gigas substitutes of Radix Angelicae in the market of South East Asia 当归 A. sinensis cultivated mainly in Minxian Gansu (岷县，甘肃省) Lao et al. (2004). Analytica Chimica Acta 526:131-137 Zhao et al. (2003). J. Agric. Food Chem. 51:2576-2583

  6. Ancient preparation (古方) : Radix Angelica Sinensis: Radix Astragali (1 : 5) Two bowls of water boiled by moderate heat to one bowl of decoction 以水两碗煎至一碗 Chinese Angelica Decoction Danggui Buxue Tang (DBT) 当归补血汤 甘肃 Gansu 山西 Shanxi 当归 Radix Angelicae 黄芪 Radix Astragali

  7. 金元四大家 Liver Wood Water Kidney Radix Astragali (King) Reinforce the Qi of Spleen and Lung 甘温之品，补脾肺之气为君 DBT Gold Earth Radix Angelica Sinensis (Minister) Nourishing the Blood and Ying Lung Spleen 甘辛而温，养血和营为臣 李东垣写道:“该方可为益气生血之剂,称之有形之血,生于无形之气,黄芪用量五倍于当归,大补脾肺元气,用当归益血和营,则阳生阴长,气旺血生”。 Li said: the visiable Blood is originated from the invisiable Qi. In DBT, RA, the amount of which is 5 times of RAS, can greatly tonify the Qi; RAS, nourish the Xue. Combined the two herbs, Yin and Yang were coordinated harmonizely and the formulation can take its effects by reinforcing Qi and promoting Xue. 李东垣 补土派 刘完素 寒凉派 朱震亨 滋阴派 张从正 攻下派

  8. Quality analyses of DBT by 2 approaches 利用化学和生物分析当归补血汤的质素 Chemical analysis 化学分析 Biological analysis 生物分析 Activation of promoter of estrogen receptor in breast cancer cells MCF-7 (乳癌細胞) Proliferation of osteosarcoma cells MG63 (骨骼细胞) Immuno-stimulating effects of T-lymphocyte (T-淋巴細胞) and macrophage (巨噬細胞) Cytokine secretion of T-lymphocyte (T-淋巴細胞) Gene expression of osteosarcomaMG63 (骨骼细胞)by microarray

  9. 0.06 0.04 0.02 0 黃芪: 当归比例 黃芪: 当归比例 (RA:RAS) (RA:RAS) Ligustilide 藁本内酯 Total Polysaccharides 总多糖 Ferulic Acid 阿魏酸 0.06 20 0.04 Ferulic Acid & Ligustilide (g/100g) Total Polysaccharides (g/100g) 10 0.02 0 0 1:1 2:1 3:1 4:1 5:1 7:1 10:1 Radix Angelica sinensis Extraction of DBT with different ratios 不同成份比例当归补血汤的提取物 Calycosin 毛蕊异黄酮 Astragaloside IV 黃芪甲苷 Formononetin 芒柄花素 Active constituent (g/100g) 1:1 2:1 3:1 4:1 5:1 7:1 10:1 Radix Astragali

  10. 0.8 A: Astragaloside IV黃芪皂苷 C: Calycosin 毛蕊异黃酮 E: Ligustilide 藁本內脂 G: Total Flavonoids 总黃酮 B: Ferulic Acid 阿魏酸 D: Formononetin 芒柄花素 F: Total Saponins 总皂苷 H: Total Polysaccharide总多糖 0.6 0.4 0.2 0 -0.2 55 5 10 15 20 25 30 35 40 45 50 55 5 10 15 20 25 30 35 40 45 50 Quality control of DBT: Determination of correlation coefficients 利用相关因子分析控制当归补血汤的质量 D MG 63 cells (MTT) MG 63 cells (ALP) D G G F A F A H C E E B H C B B 0.6 Platelet MCF-7 cells C G 0.4 A A F D H 0.2 C F E 0 B D E -0.2 G H P < 0.01 P > 0.01

  11. 内外伤辩惑论 Ancient preparation (古方) : Radix Angelica sinensis 当归: Radix Astragali 黄芪 (1 : 5) Two bowls of water boiled by moderate heat to one bowl of decoction 以水两碗煎至一碗 1:5 ratio showed the highestamount of active constituent except Ligustilide 除了藁本内酯, 当归:黄芪(1:5)煎煮, 相对不同的比例, 能够增加当归和黄芪不同的有效成分. Song et al. (2004). Planta Medica 70: 1222-1227 Dong et al. (2006). J. Agric. Food Chem. 54: 2767-2774 Gao et al. (2006). FEBS Letter (in press) Gao et al. (2006). Planta Medica 72: 1227-1231

  12. Effect of different ratio DBT on estrogen-like response in MCF-7 cells and proliferation, differentiation responses in MG-63cells 当归补血汤 (不同比例) 对女性荷尔蒙受体的基因调控和骨细胞生长及分化的反应 30 DBT Estrogen Estrogen-like substance(s) From DBT 20 * Estrogen receptor 10 Transcription 0 Light 30 Luciferase proteins Lucifercin Estrogen-like response 100 20 20 10 10 0 1 : 1 2:1 3:1 4:1 5:1 7:1 10:1 0 Luciferase Assay Luciferase Assay ERE ERE Luciferase Osteosarcoma MG-63 cell Breast cancer MCF-7 cell Proliferation Cell viability (% of increase) Differentiation * ALP activity (% of increase) Luciferase activity (% of increase) 1 : 1 2:1 3:1 4:1 5:1 7:1 10:1 Radix Angelica sinensis Radix Astragali ß-Estradiol Different Extraction Group 黄芪:当归比例 (RA:RAS) Radix Angelica sinensis Radix Astragali ß-Estradiol Different Extraction Group 黄芪:当归比例 (RA:RAS) Dong et al. (2006). J. Agric. Food Chem. 54: 2767-2774

  13. Estrogen induces the phosphorylation ofSer 118 and Ser 167 of estrogen receptor in MCF-7 cell 女性荷尔蒙对女性荷尔蒙受体a丝氨酸118 和 167的影响 Estrogen Other compounds Plasma membrane Raf Estrogen receptor Estrogen receptor MEKs Erk 1/2 P Ser 118 Ser 167 Rsk P Sp1 Elk1 NF-κB c-Fos c-Jun bind to estrogen-responsive element (ERE) on promoter of target gene phosphorylated by other signaling pathways, e.g MAP kinase-dependent pathway, to mediate the gene transcription. 3 Luciferase activity (Fold of control ) 2 ERE/ERE 1 0 0 1 2 3 4 0.5 0.25 0.125 DBT concentration (mg/ml) Estrogen Estrogen Ras Estrogen receptor Estrogen receptor Gene Expression

  14. DBT induces the phosphorylation of estrogen receptor aon Ser-118 in MCF-7 cells 当归补血汤对女性荷尔蒙受体a 丝氨酸118的磷酸化效应 0 10 20 30 40 50 Control 3 DBT Radix Angelica Radix Astrgali 17β- estradiol RAS + RA 2 Estrogen Receptor a 1 Ser-118- P 0 0 10 20 30 40 50 bind to estrogen-responsive element (ERE) on promoter of target gene - ICI 182780 8 + ICI 182780 6 0 20 40 60 0 20 40 60 Amount of ER αS118-P ( x Basal ) 4 2 0 Control DBT 17β- estradiol Min ERαS118-P Control ERα Other compounds ERαS118-P DBT ERα ERαS118-P Radix Angelica (RAS) Amount of ERαS118-P ( x Basal ) ERα ERαS118-P RadixAstragali (RA) ERα ERαS118-P RAS + RA ERα ERαS118-P 17β- estradiol ERα Min No pretreatment ICI pretreatment Min ERαS118-P Control ERα ERαS118-P DBT ERα ERαS118-P 17β- estradiol ERα

  15. DBT, but not estrogen, induces the phosphorylation of estrogen receptor aon Ser-167 in MCF-7 cells 当归补血汤对女性荷尔蒙受体a 丝氨酸167的磷酸化效应 Control 17β- estradiol Other compounds DBT Radix Angelica Radix Astrgali RAS + RA Estrogen Receptor a Ser-167- P 20 30 40 50 10 0 phosphorylated by other signaling pathways, e.g MAP kinase-dependent pathway, to mediate the gene transcription. - ICI 182780 + ICI 182780 0 20 40 60 0 20 40 60 Min 0 10 20 30 40 50 ERαS167-P 15 Control ERα ERαS167-P DBT ERα 10 Radix Angelica (RAS) ERαS167-P Amount of ERαS167-P ( x Basal ) ERα Radix Astragali(RA) ERαS167-P ERα 5 ERαS167-P RAS + RA ERα ERαS167-P 0 17β- estradiol ERα Min No pretreatment ICI pretreatment 12 Min ERαS167-P Amount of ERαS167-P ( x Basal ) 8 Control ERα 4 ERαS167-P DBT ERα 0 Control DBT

  16. 12 8 4 0 - ICI 182780 + ICI 182780 0 0 15 15 30 30 45 45 Erk1/2 P Erk 1/2- P Erk 1/2 DBT induces the phosphorylation of Erk1/2 in MCF-7 cells 当归补血汤对乳腺癌细胞外信号调节激酶的磷酸化效应 Min 15 Control 0 15 30 45 Other compounds DBT Erk 1/2-P RAS + RA Control Erk 1/2 10 Amount of ERK 1/2- P ( x Basal ) Erk 1/2- P DBT 5 Erk 1/2 MAP kinase- dependent pathway Erk 1/2- P RAS + RA 0 Erk 1/2 0 15 30 45 Min No pretreatment ICI pretreatment Min Erk 1/2- P Amount of ERK 1/2- P ( x Basal ) Control Erk 1/2 DBT Control DBT

  17. 12-O-Tetradecanoyl-phorbol 13-Acetate (TPA) induces the phosphorylation of Ser-118 and Ser-167 of estrogen receptor in MCF-7 cells Min: 0 15 30 45 Erk1/2-P Control Erk1/2-P ER S118-P Erk1/2 ER S167-P Erk1/2-P TPA Erk1/2 15 ER S118-P Control Phosphorylation ( x Basal ) Ser-167- P Ser-118- P ER 10 ER S118-P TPA ER 5 ER S167-P Control ER 0 0 15 30 45 ER S167-P TPA Min ER 四葵酸佛波乙酯对乳腺癌细胞的女性荷尔蒙受体α丝氨酸118和167的磷酸化效应 TPA PKC Raf MEK1/2 Erk1/2 Estrogen Receptor a

  18. The DBT-induced phosphorylation of Erk, estrogen receptor (S-118 and S-167) are blocked by U0126 and PD98059 in MCF-7 cells PD98059, U0126有效阻挡当归补血汤引發乳腺癌细胞的外信号调节激酶, 女性荷尔蒙受体α丝氨酸118和167的磷酸化效应 DBT Erk1/2-P Control Raf Erk1/2-P DBT 8 ER S118-P Control ER S118-P DBT MEK1/2 4 ER S167-P Control ER S167-P U0126 20μM DBT Erk1/2 0 Ser-118- P Ser-167- P DBT DBT DBT Control Control Control Estrogen Receptor a PD98059 20μM Pretreated with : PD098059 Buffer U0126 Min: 0 30 0 30 0 30 ER S118-P ER S167-P Erk1/2-P Buffer PD098059 U0126 Phosphorylation ( x Basal ) ** ** ** **

  19. DBT in different ratios induces the proliferation and IL-2 secretionof T-lymphocytes 当归补血汤对T-淋巴细胞生长和分泌IL-2的影响 IL-2 secretion proliferation 140 ** 140 ** 40 60 Proliferation (% of increase) IL-2 (pg) 40 20 20 0 0 1:1 3:1 5:1 7:1 10:1 1:1 3:1 5:1 7:1 10:1 PHA PHA Radix Astragali Radix Astragali Radix Angelicae Sinensis Radix Angelicae Sinensis Different Extraction Group 黄芪:当归比例 (RA:RAS) Different Extraction Group 黄芪: 当归比例 (RA:RAS) Gao et al. (2006). Planta Medica 72: 1227-1231

  20. DBT induces T-lymphocyte secreted cytokines Cytokines detected IL-2 IL-10 IL-13 IL-4 TNF-a IL-5 INF-g IL-6 IL-8 Gm-CSF 当归补血汤对T-淋巴细胞分泌不同白介素的影响 IL-2 120 Antibody microarray: Quick and accurate to quantify the antibody secretion by T-lymphocyte. IL-6 110 IL-8 6 IL-10 5 Cytokine secreted (x Basal) 4 3 2 1 0 PBS RAS RA RAS + RA DBT

  21. P P P P P P P IL-2 inducesthe signaling mechanism in T-lymphocytes IL-2对T-淋巴细胞信号机理的影响 IL-2 β γ α IL-2 receptor GTP RAS GDP P ERK RAS MEK P RAF1 RAF1 P MEK ERK STAT c-FOS c-JUN c-FOS Elk-1 Elk-1 Gene expression

  22. Control DBT Radix Astragali Radix Angelicae RAS + RA Erk1/2 P DBT induces the phosphorylation of Erkin T- lymphocytes 当归补血汤对T-淋巴细胞外信号调节激酶的磷酸化效应 Min 0 10 20 30 45 60 120 Erk 1/2-P Erk 1/2-P Control (x Basal) Erk 1/2 8 Erk 1/2-P Radix Astragali Erk 1/2 ERK1/2 6 Radix Angelicae Erk 1/2-P Erk 1/2 4 p Erk 1/2-P DBT Erk 1/2 2 Erk 1/2-P Amount of DG+HQ 0 Erk 1/2 20 40 60 80 100 120 0 Erk 1/2-P PMA Min Erk 1/2 Other compounds IL-2 receptor Raf Ras MEK1/2 Elk Gao et al. (2006). Planta Medica 72: 1227-1231 IL-2

  23. Microarray gene analysis after treatment of DBT, DG and HQ in MG63 cells 当归 黄芪 当归补血汤专属 利用基因芯片技术研究当归补血汤,当归与黄芪对骨骼细胞的影响 Microarrays Control Radix Angelica Sinensis Radix Astragali DBT 对照 当归 黄芪 当归补血汤 - Samples Among total 7488 genes on the chip, several genes related to different systems were shown to be up-regulated only in DBT treatment, but not in DG or HQ treatments Up or down regulated by Radix Angelica Sinensis (当归) 当归补血汤 1063 genes Up or down regulated byRadix Astragali (黄芪) 黄芪 当归 661 genes Up or down regulated by DBT (当归补血汤) 884 genes

  24. Micro-array analysis of DBT, RAS and RA in MG63 cells 利用基因芯片技术研究当归补血汤,当归与黄芪对骨骼细胞的影响 Gene 当归 黄芪 当归补血汤 女性荷尔蒙 IFI-6-16 0.5 2.54 21.8 32.58 IFN RF-3 1.8 3.4 108.5 152.5 Immune system IL-1a 1.6 2.2 5.3 8.9 COL2A1 0.13 0.8 3.2 0.46 ECM & Proteases MMP1 0.43 1.7 2.9 0.59 H2AFA 2.78 2.14 3.74 2.32 H2AFL 2.01 1.42 2.43 1.76 Gene transcription H2AFO 2.67 1.87 3.97 2.56 GFER 2.19 2.57 3.17 1.59 TNFSF7 1.35 1.4 9.56 2.16 Cell cycle & Apoptosis IER3 3.66 6.37 10.19 5.04 CCL2 2.35 0.87 88.22 1.00 CCL7 0.33 5.96 175.02 2.21 Bone development & differentiation CCL8 2.73 2.89 184.65 1.09 Gal-9 1.6 1.77 441.76 0.73 Ratio = Treatment / Control

  25. RNA extraction RT-PCR analysis Gene validation Method Treatment Control RAS DBT MG63 cells RA CCL2 CCL7 600 CCL8 300 CCL2 CCL7 CCL8 200 Gal-9 Gal-9 GAPDH 100 GAPDH 0 CCL2,7,8 (Chemokine 2,7,8)  a family of small cytokines for chemoattraction of white-blood-cells Gal-9 (Galectin 9) b-galactoside-binding proteins for cell-cell and cell-matrix interaction  Both of these genes are shown to involve bone development and differentiation Validation of gene expression by RT-PCR analysis 利用反转录酶-聚合酶链式反应法确认当归补血汤对骨骼细胞基因的表现 ** Amount of mRNA ( % of control ) Control DBT RA RAS

  26. Conclusion 结论 The sources of Huangqi and Danggui, based on the amount of chemical constituents, have been determined. 测定了不同产地黄芪, 当归中各化学组分的含量。 Astragaloside IV, ferulic acid, calycosin, formononetin, ligustilide, total saponins, total flavonoids and total polysaccharide could be used as chemical marker in DBT。 黄芪皂苷,阿魏酸,毛蕊异黄酮,芒柄花素, 藁本内脂, 总皂苷, 总黄酮和总多糖可以作为当归补血汤的化学指针。 Different bio-assays were established, which could be used for chemical markers of Huangqi, Danggui and DBT. 通过不同的生物活性实验, 可以用于评价黄芪, 当归及当归补血汤的化学指针。 The optimized extraction condition of DBT has been determined, which is in line with the ancient formulation. 优化了当归补血汤的提取工艺, 结果与古方一致。 Search for DBT-specific biomarkers: Proteomic and gene analysis 运用类蛋白体技术和基因研究寻找当归补血汤专属的生物指标。

  27. 谢谢

  28. Micro-array analysis of DBT, DG and HQ in MG63 cells 利用基因芯片技术研究当归补血汤,当归与黄芪对骨骼细胞的影响 Gene 当归 黄芪 当归补血汤 女性荷尔蒙 IFI-6-16 0.5 2.54 21.8 32.58 IFN RF-3 1.8 3.4 108.5 152.5 Immune system IL-1a 1.6 2.2 5.3 8.9 COL2A1 0.13 0.8 3.2 0.46 ECM & Proteases MMP1 0.43 1.7 2.9 0.59 H2AFA 2.78 2.14 3.74 2.32 H2AFL 2.01 1.42 2.43 1.76 Gene transcription H2AFO 2.67 1.87 3.97 2.56 GFER 2.19 2.57 3.17 1.59 TNFSF7 1.35 1.4 9.56 2.16 Cell cycle & Apoptosis IER3 3.66 6.37 10.19 5.04 CCL2 2.35 0.87 88.22 1.00 CCL7 0.33 5.96 175.02 2.21 Bone development & differentiation CCL8 2.73 2.89 184.65 1.09 Gal-9 1.6 1.77 441.76 0.73 Ratio = Treatment / Control

  29. 当归 黃芪 当归补血汤 The HPLC profile of DBT, DG and HQ 利用液相色谱研究当归补血汤,当归与黄芪 HPLC Conditions： Flow rate: 1 ml/min Wavelength: 254 nm Column: Waters DeltaPak C18 (3.9*150 mm, 5 mm) Mobile Phase: A: Acetonitrile, B: 0.01% Phosphoric acid (0 min A 15%, B 85% : 60 min A 65%, B 35%)

  30. 120 100 80 60 40 20 0 Different DBT fractions induce T-lymphocyte proliferation 不同当归补血汤提取物对T-淋巴细胞生长的影响 Fractionation of DBT by size and nature T-lymphocyte proliferation assay DBT % of increase Centrifugation Ethanol precipitation <10 kDa Polysaccharide- enriched >10 kDa Polysaccharide- depleted DBT PHA >10 kDa <10 kDa Polysaccharide- depleted Polysaccharide- enriched 100 mg/ml of fraction

  31. DBT-specific regulated genes interferon, alpha-inducible protein (clone IFI-6-16) IFI-6-16 IFN RF-3 interferon regulatory factor 3 small inducible cytokine A7 (monocyte chemotactic protein 3) Cytokine A7 interleukin 1, alpha IL-1a H2AFA H2A histone family, member A Immune system H2A histone family, member L H2AFL H2A histone family, member O H2AFO COL2A1 collagen, type II, alpha 1 matrix metalloproteinase 1 MMP1 GFER growth factor, erv1 TNFSF7 tumor necrosis factor IER3 immediate early response 3 当归补血汤专属调节的基因 Transcription ECM & proteases Cell cycle & apoptosis

  32. Proteomics analysis of human breast cancer MCF-7 cells Sample Preparation pH3 pH10 - + 运用类蛋白体技术研究当归补血汤对乳腺癌细胞的效应 MCF-7 当归 黄芪 当归补血汤 Imaging & Analysis Identification 2-D Electrophoresis

  33. Analysis of 2D gel electrophoresis after treatment of DBT, DG and HQ in MCF-7 cells 运用2D电泳检定法研究当归补血汤,当归与黄芪对乳腺癌细胞的影响 当归补血汤 对照 当归 黄芪

  34. DBT induces T-lymphocyte secreted cytokines IL-2 120 IL-6 110 IL-8 6 IL-10 5 Cytokine secreted (x Basal) 4 3 2 1 0 PBS RAS RA RAS + RA DBT Different DBT samples (100 μg/ml) IL-2 is necessary during T cell development in the thymus for the maturation, T cell immunologic memory. IL-6 is a pro-inflammatory cytokine secreted by T cell, osteoblast secrete IL-6 to stimulate osteoclast formation and bone remodeling. IL-8 is a pro-inflammatory cytokine produced by T cell, which acts on neutrophils as a chemoattractant, activator, and modulator of endothelial adhesion and transmigration. IL-10 known as human cytokine synthesis inhibitory factor (CSIF), is an anti-inflammatory cytokine, capable of inhibiting synthesis of pro-inflammatory cytokines. 当归补血汤对T-淋巴细胞分泌不同白介素的影响 Cytokines detected IL-2 IL-10 IL-13 IL-4 TNF-a IL-5 IL-6 INF-r IL-8 Gm-CSF

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