The vasorelaxation effect of an extract of Chinese medicinal herb, Radix Angelica Pubescentis,

1. 2006 World Congress on Chinese Medicine The vasorelaxation effect of an extract of Chinese medicinal herb, Radix Angelica Pubescentis, in porcine coronary artery 獨活提取物對豬冠狀動脈血管舒張的影響 Institute: The University of Hong Kong Supervisor: Prof. Ricky Y. K. Man Prof. C. M. Che Presenter: Matthew L. Y. Chan

2. Introduction Chinese Medicinal Herb Pharmaceutical name: Radix Angelicae Pubescentis Latin botanical name: Angelica pubescens Maxim. f. biserrata Shan et Yuan Chinese (English) name: 獨活(Du Huo)

3. Introduction extract ingredients * # Pharmacological Actions: Chinese Pharmacopoeia Recent Publications • anti-inflammatory*# • anti-cancer*# • vaso-relaxant # • anti-oxidant # • anti-arthritis • headache • thrombosis • vessels dilation • promotion blood circulation

4. Introduction Osthole (0.5%) Xanthotoxin • anti-arrhythmic • vaso-relaxant • antioxidant Angelmarin • anti-cancer Major active ingredients

5. Material Dry AP samples Powdered herb 20 mg/ml APE Grind Osthole powder Store at -20˚C and dissolve in DMSO before use Extract trice with 95% ethanol (1:5, w/v) (stir overnight, centrifuge and evaporate to dryness) 10 mM Osthole Store at -20˚C and dissolve in DMSO before use Crude extract Preparation of APE and osthole

6. Method Amplifier Force transducer Organ bath Circulator Organ bath setup

7. Method Measurement of isometric tension change Krebs-Henseleit solution (KHS) (NaCl 120 mM, KCl 4.76 mM, NaHCO3 25 mM, NaH2PO4·2H2O, 1.18 mM CaCl2 1.25 mM, MgSO4·7H2O 1.18 mM, D-glucose 5.5 mM) Water out Water in 95% O2 5% CO2 3 mm coronary arterial ring Preparation of the porcine coronary arterial rings

8. Method Viability test Tension Cumulative addition of drug > 80 % relaxation > 4 g contraction Wash with Krebs 2 g resting tension Time < incubation > U46619 (30 nM) Indo (10 μM) Indo and/or blocker BK (1 μM) U46619 (30 nM) Schematic diagram of organ bath technique

9. Results & Discussion Part I A B Endothelium-intact C D Endothelium-disrupted Endothelium-disrupted Response to APE on porcine coronary arterial rings

10. Conclusion Part I APE L-arginine NO eNOS - Endothelium PKG GC cGMP GTP + Relaxation Vascular smooth muscle cell L-NAME Schematic diagram depicts the proposed mechanisms of APE Endothelium-intact 

11. Conclusion Part I APE ODQ SQ22536  APE - -      AC GC - cAMP cGMP GTP - + ATP KT5720 Rp-8-Br-cGMPS + - Rp-8-Br-cAMPS - PKA PKG KT5823 Vascular smooth muscle cell Relaxation Schematic diagram depicts the proposed mechanisms of APE Endothelium-disrupted

12. Results & Discussion Part II E ∗ p < 0.05 versus without EC group * G F Endothelium-intact Endothelium-intact ∗ p < 0.05 versus L-NAME group ∗ p < 0.05 versus ODQ group * * Response to osthole on porcine coronary arterial rings

13. Results & Discussion Part II E ∗ p < 0.05 versus without EC group * H Endothelium-disrupted Endothelium-disrupted I Response to osthole on porcine coronary arterial rings

14. Conclusion Part II Osthole (10 μM) L-arginine NO eNOS - (10 μM) Endothelium - PKG GC cGMP GTP + Relaxation Vascular smooth muscle cell ODQ L-NAME Schematic diagram depicts the proposed mechanisms of osthole Endothelium-intact

15. Conclusion Part II Osthole   - - AC GC cAMP cGMP GTP + ATP + PKA PKG Vascular smooth muscle cell Relaxation ODQ SQ22536 Schematic diagram depicts the proposed mechanisms of osthole Endothelium-disrupted

16. Future Plan • Continue to study the mechanism of action APE work on, such as K and Ca channel. • Continue to study the mechanism of action osthole work on, such as those downstream pathway of cGMP cascade.

18. Calculated concentration of osthole in APE Actual concentration of osthole Comparison of the osthole concentration used in the experiment