1. Flavonoids from wild Aquilaria sinensis leaves —— biological activities and regulation effects against the impairments of Aquilaria sinensis plants Speaker: Dr. Heru Chen College of Pharmacy Jinan University Guangzhou, P. R. China

2. Introduction Results/Discussion Conclusion Acknowledgement

3. Zhongshan city

5. Introduction • Eaglewood is the materials from the impaired response of Aquilaria sinensis (Lour.) Gilg。Its productive process is quite accidentaland time-consuming. Therefore, eaglewood is very expensive and scarce.However, leaves are rich and reproducible. • Extract from the leaves has been indicated with activities of sedative, anti-inflammatory, anti-oxidation, and anti-cancer. The main components were reported as flavones and flavonoids • Some flavonoids may involve in the productive process of eaglewood.

6. Introduction Results/discussion Conclusion Acknowledgement

7. Results and discussion • Procedure applied

8. Results and discussion • Flavonoids isolated Y4 Y7 Luteolin-7, 3', 4'-trimethyl ether Apiolin-7, 4'-dimethyl ether Y9 Y8 5,4’-dihydroxy-7,3’-dimethoxyflavone Genkwanin

9. Results and discussion • Flavonoids isolated Y18* Y19* Quercetin-3-O--D- galactopyranoside 6-hydroxy-7, 4’-dimethoxyflavone Y21* Y22 5,4'-dihydroxy-7-methoxyflavanone 5-hydroxy-7, 2', 4', 5'- tetramethoxyflavone

10. Results and discussion • Flavonoids isolated Y24 Y23 Quercetin Kaempferol Y27 Quercetin-3-O--D-glucopyranoside

11. Results and discussion • Flavonoids isolated Dihydroflavonoid firstly isolated from Aquilaria！ *compoundsfirstly isolated from Thymelaeaceae # compounds firstly isolated from Aquilaria of Thymelaeaceae

12. Results and discussion • Anti-cancer activities of flavonoids MTT assay was applied “-” means no data

13. Results and discussion • Anti-cancer activities of flavonoids MTT assay was used A: acetone P: petroleum E: ethanol (in the front) E: EtOAc (back) M: methanol B: butanol

14. Results and discussion • Clearance effect against nitrite (NO2-) pH 3.0, 37C

15. Results and discussion • Clearance mechanism against nitrite (NO2-) 2 1 3

16. Results and discussion • Clearance mechanism against nitrite (NO2-)

17. Results and discussion • Regulation of flavonoids against plant impairments 1 2 3 4 6 5 Aquilana sinensis 7 8 9 10 11

18. Results and discussion • Regulation of flavonoids against plant impairments Normal Drilling+H2O Drilling+DMSO Drilling+DMSO +flavonoids

19. Results and discussion • Regulation of flavonoids against plant impairments CK2: drilling+DMSO CK1-a: normal plant CK1-b: drilling+H2O

20. Results and discussion • Regulation of flavonoids against plant impairments G0.1: drilling+0.1 mg/ml Genkwanin in DMSO G0.3: drilling+0.3 mg/ml Genkwanin in DMSO G0.5: drilling+0.5 mg/ml Genkwanin in DMSO G0.8: drilling+0.8 mg/ml Genkwanin in DMSO G1.0: drilling+1.0 mg/ml Genkwanin in DMSO

21. Results and discussion • Regulation of flavonoids against plant impairments almost showed no effect in day 1 A0.1: drilling+0.1 mg/ml Apiolin in DMSO A0.5: drilling+0.5 mg/ml Apiolin in DMSO A1.0: drilling+1.0 mg/ml Apiolin in DMSO Q: Quercetin K: Kaempferol

22. Results and discussion • Regulation of flavonoids against plant impairments Q0.1 G0.5 day 1 day 5 Y(II): effective quantum yield of photosystem II Photosynthetic rate decreases after day 5.

23. Results and discussion • Regulation of flavonoids against plant impairments Q1.0 day 1 day 5 Production of proline is a kind of response of plant against adversity

24. Results and discussion • Regulation of flavonoids against plant impairments day 1 day 5 Production of malondialdehyde (MDA) is a kind of response of plant against adversity (oxidative stress)

25. Results and discussion • Regulation of flavonoids against plant impairments G0.5 day 1 day 5 Production of phenylalanine amonialyase (PAL) is a kind of response of plant against impairments

26. Results and discussion • Regulation of flavonoids against plant impairments Acacetin Genkwanin Apiolin-7,4‘-dimethyl ether Luteolin-7,3',4‘-trimethyl ether

27. Results and discussion • Regulation of flavonoids against plant impairments total proteins from day 1 leaves  2-DE  analysis of differential proteins  confirmation of the point with differential proteins  enzyme digestion within the gel  Analysis of Mass spectra  compared with the database and confirmed proteins

28. Results and discussion • Regulation of flavonoids against plant impairments 3D map of differential proteins. 1： CK2 group; 2: G0.5 group

29. Results and discussion • Regulation of flavonoids against plant impairments

30. Results and discussion • Regulation of flavonoids against plant impairments AP-Promoter defensive substance PT/ST Promoter DB/NA Promoter Thylose TC/MG Promoter external impairment Lignans phytoalexin MT Promoter AP: autoplast protein; PT: protein transportation ST: signal transduction; DB: DNA bonding NA: nucleosome assembly; TC: translation control MG: microbial growth; MT: methyl transferase

31. Introduction Results/discussion Conclusion Acknowledgement

32. Conclusion 11 flavonoids were obtained the leaves of wild type Aqualaria plant. 5 flavonoids were demonstrated good inhibitory activity against DU145、 PC-3, and HepG2 cancer cell lines, respectively. Compound Y1 (mangiferin) and Y2 (2-O-α-L-rhamnopyranosyl-4, 6, 4'-trihydroxybenzo-phenone) were identified as the two most active compounds with scavenging rate 15.33%±0.81% and 14.87 %±0.38% at the conditions of pH 3, 37℃, and 30 min duration, respectively. Genkwanin with moderate concentration (0.5mM) exhibited significant protective effect against mechanical and chemical injury to Aquilaria sinensis plants.

33. Introduction Results/discussion Conclusion Acknowledgement

34. Acknowledgement

35. Acknowledgement  National Natural Science Foundation of China (No. 81172982)  Key projects of social development of science and technology project in Guangdong Province (No. 2010A030100006)

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