**4. Conclusion**

254 Rheumatoid Arthritis – Treatment

Table 13. Protein carbonyls, HNE (4-hydroxynonenal) and MDA (malondialdehyde)-protein adducts levels in plasma measured on experimental day 28 in an experiment with coenzyme Q10 (CoQ) and methotrexate (MTX) evaluated as monotherapy and combination therapy (CoQ+MTX). Changes in all groups with arthritis were calculated compared to the control value assessed for healthy control animals on experimental day 28. For statistical analysis of

**Parameters CO AA AA-CoQ AA-MTX AA-CoQ** 

\*\*

\*\*\*

Table 14. GGT (-glutamytransferase) activity in joint, IL-1α (interleukin-1) and CoQ9 (coenzyme Q9) levels in plasma measured on experimental day 28 in an experiment with coenzyme Q10 (CoQ) and methotrexate (MTX) evaluated as monotherapy and combination

The functionality of peripheral blood neutrophils in AA was evaluated by phagocytosis,

Both phagocytosis and oxidative burst were increased due to arthritis. The immunosuppressive effect of MTX was demonstrated in lowering all characteristics of the functionality of peripheral blood neutrophils, not only in comparison with arthritis but also with CO. The addition of CoQ10 to MTX modulated all processes back to the level of CO. The observed immunoenhancing activity of CoQ10 may prove beneficial in MTX routine treatment. In this experiment, flow cytometric determination of the functionality of

9.38±0.76 16.08±0.96

therapy (CoQ+MTX). For statistical analysis of data see table 1.

neutrophils was first applied for an experimental model on rats.

**HNE-protein adducts** 

125.80±8.74 \*\*\*

111.31±7.85 +

91.66±8.571 +++ / #

67.76±13.59 38.27±4.21

16.16±1.07 6.83±1.21

\*\* 241±21.28 189±16.51 209±21.75 158±12.65

++

+++

114.03±9.41 112.92±4.49

**MDA-protein adducts** 

131.56±13.13 \*\*\*

101.49±7.45 ++

85.376±5.129 +++ / # # #

**+MTX** 

14.83±0.90 ++ / # #

9.60±1.05 +++

+

**Protein carbonyls** 

\*\*\*

+

+++

+++

**AA** 189.49±3.19

**AA-CoQ** 169.74±3.19

**AA-MTX** 146.50±4.78

**AA-CoQ+MTX** 122.93±3.886

**(pg/ml)** 42.95±7.88 87.11±8.52

154±7.70

oxidative burst and metabolic activity (Table 15).

**Oxidative stress parameters (%)** 

data see table 1.

**IL-1α**

**Activity of GGT in joint (†)** 

> **CoQ9 in plasma (nmol/l)**

† - nmol 4-nitroaniline /min /g tissue

In the past our research team, using the AA model, has monitored OS and inflammation in time course using different clinical and biochemical/immunological markers, and at the same time we have assessed the efficacy of the administered experimental substances with regard to their ability to reduce OS and inflammatory processes. In our experiments on AA rats we observed a beneficial effect of administration of low molecular weight antioxidants (coenzyme Q and carnosine), high molecular weight immunomodulators/antioxidants (glucomannan, Imunoglukan®) and compounds related to plants (curcumin, arbutin, pinosylvin, sesame oil, and extracts from *Boswellia serrata*, *Arctostaphylos uva-ursi* and *Zingiber officinale*).

In light of these results, we proceeded in the search for the most suitable therapeutic substance (an antioxidant/immunomodulator) with the ability to improve the therapy of RA with MTX. The aim was to find a potential enhancement of the antirheumatic effect of MTX in particular combinations compared to monotherapy. Carnosine, coenzyme Q, pinosylvin and Imunoglukan® were selected for assessment of a combinatory therapy with MTX. The already performed experiments on arthritic rats with pinosylvin, Imunoglukan® and coenzyme Q confirmed the hypothesis about the beneficial effect of adding a suitable immunomodulator/antioxidant to the therapy with MTX. Final safety and efficacy of these approaches calls for further more detailed research not only in preclinical but also in clinical conditions.
