The unit provides a range of laboratory capabilities, and has the facilities to determine the influence of natural products on numerous physiological markers. This includes measuring the effects on immune function, anti-inflammatory response, platelet aggregation and cell cycling by flow cytometric techniques using both fresh human blood and multiple cultured cell lines; or by measuring the effects in vitro on anti-inflammatory markers, antioxidant capacity, cholesterol synthesis, oestrogen synthesis, and glucose metabolism, as well as determination of the cytotoxicity of natural products. Being part of CPP provides expertise in sample preparation and chemical analysis of natural products. CPP and the pharmacology unit actively develop new methods of analysis, and clients/students are encouraged to discuss their project with staff, even if they think it is outside the current capabilities of the unit.

PHARMACOLOGY RESEARCH CAPABILITIES

Antioxidant Capacity
The antioxidant capacity of agents is tested using the Oxygen Radical Absorbance Capacity method.

Cell Proliferation, Cell Death and Cytotoxicity
In-vitro cell proliferation, cell death and/or cytotoxicity is investigated by using the following assays
• Inhibition of ATP production
• Apoptosis (using Annexin V)

A range of cell lines is available for testing the cytotoxicity of natural products.

Anti-cancer Properties
The potential for agents to act as anti-cancer drugs is tested in vitro, utilising cell cycle analysis. This method can also be used for measuring DNA content (ploidy levels).

Platelet Aggregation
The ability of natural products to reduce platelet aggregation in vitro is tested using a whole blood model and analysed by flow cytometry. Platelet aggregation is induced by the agonists ADP and collagen.

Inflammation
A broad range of diseases are associated with an inflammatory process. The following inflammatory markers allow assessment of the progress of the inflammatory process and the efficacy of anti-inflammatory agents.
• Interleukins
• Tumour Necrosis Factor Alpha
• High sensitivity C-Reactive Protein
• Matrix metalloproteinases
• Cyclyoxygenase 1
• Cyclooxygenase 2
• Prostaglandin E2
• Cytokines (IL-8, IL-1B, IL-6, IL-10, TNFa, IL-12p70))
• Nitric Oxide
• Nuclear factor ? B

Immune Function
Using flow cytometry, non-specific and specific immune responses can be measured using these assays
• Phagocytosis of granulocytes and monocytes
• Oxidative burst of granulocytes and monocytes
• Natural killer cell cytotoxicity
• Lymphocyte activation (Total T, T Helper & T Suppressor)
• Cytokine production (TH1/TH2): IL-2, IL-4, IL-5, IL-6, IL-10, TNFa, IFN
• Lymphocyte subsets (Total T, Helper T, Suppressor T, B & NK cells)

Glucose Metabolism Inhibition
The influence of natural products on the glucose index is tested by measuring their ability to inhibit glucose metabolic enzymes, including
• a-amylase
• a-glucosidase

Inhibition of Cholesterol Synthesis
Inhibition of cholesterol synthesis is measured in vitro using the cell line HepG2. The protocol was developed within the unit, whereby reduced uptake of C14-acetic acid from culture media suggests an inhibition of cholesterol synthesis.