Antioxidant and Anti-Inflammatory Activity of Pine Pollen Extract

Antioxidant and Anti-Inflammatory Activity of Pine Pollen Extract In Vitro

Authors: Kyung-Hee Lee, Ae-Jung Kim, Eun-Mi Choi

Introduction

Free radicals are implicated in various diseases and health conditions, including coronary heart disease, inflammation, stroke, diabetes mellitus, rheumatic disease, liver disorders, renal failure, cancer, and aging. Inflammatory cells like polymorphonuclear leukocytes generate and release reactive oxygen species (ROS) and reactive nitrogen species (RNS), causing cellular and tissue damage and augmenting inflammation. This study investigates the antioxidant and anti-inflammatory activities of pine pollen extract (PPE) in vitro.

Objective

To determine the medicinal properties of pine pollen, focusing on its antioxidant and anti-inflammatory activities.

Methodology

Preparation of Extract

• Pine pollen (Pinus densiflora) was collected and identified.
• The dried pollen was extracted with 70% ethanol at room temperature for 3 days, concentrated, and freeze-dried.

Radical Scavenging Effect

The scavenging effects on DPPH and H2O2 were measured using spectrophotometric methods.

Antioxidant Activity in Linoleic Acid Peroxidation System:

• Measured by the ferric thiocyanate method.
• Lipid peroxides and protein oxidation in mouse liver homogenates were assessed.

Reducing Power

• Determined using Oyaizu and phosphomolybdenum methods.

Measurement of Nitrite and Cytokines in RAW264.7 Cells

• Assessed using the Griess reagent and cytokine enzyme immunoassay systems.

Determination of MMP-1 and MMP-3 Activity in SW982 Cells

• Evaluated using ELISA kits.

Detection of MAPKs Activation in SW982 Cells

• Analyzed using a MAPK ELISA kit.

Results

Radical Scavenging Effect

• PPE showed a high scavenging capacity for DPPH and H2O2 radicals, suggesting it is an effective radical scavenger.

Antioxidant Activity

• PPE exhibited significant antioxidant activity in the linoleic acid peroxidation system, comparable to α-tocopherol.
• PPE significantly inhibited lipid peroxides and protein oxidation in liver homogenates.

Reducing Power

• PPE showed high reducing power, indicating its potential antioxidant activity.

Anti-Inflammatory Activity

• PPE inhibited the production of nitric oxide (NO), TNF-α, IL-1β, and IL-6 in LPS-stimulated RAW264.7 cells.
• PPE reduced IL-1β-induced MMP-1 and MMP-3 production and JNK activation in SW982 cells.

Research Significance

This study underscores the significant potential of pine pollen extract as a natural intervention for oxidative stress and inflammatory conditions. The findings show that pine pollen extract:

• Reduces Oxidative Stress: By scavenging free radicals and inhibiting lipid peroxidation and protein oxidation.
• Enhances Antioxidant Defense: Through high reducing power and antioxidant activities.
• Mitigates Inflammation: By inhibiting the production of pro-inflammatory mediators such as NO, TNF-α, IL-1β, and IL-6.
• Suppresses Matrix Degradation: By reducing IL-1β-induced MMP-1 and MMP-3 production and JNK activation, which are involved in cartilage degradation in inflammatory diseases like arthritis.

These results pave the way for future research to explore the therapeutic applications of pine pollen extract in human subjects, potentially contributing to the treatment and prevention of oxidative stress and inflammation-related diseases.

Conclusion

Pine pollen extract demonstrates strong antioxidant and anti-inflammatory activities, suggesting its potential as a natural remedy for oxidative stress-related diseases and inflammatory conditions. These findings support the development of new herbal medicines using pine pollen extract for clinical use.

Citation

Lee, K.-H., Kim, A.-J., & Choi, E.-M. (2008). Antioxidant and anti-inflammatory activity of pine pollen extract in vitro. Phytotherapy Research, 23(1), 41–48. DOI: 10.1002/ptr.2525