Effects of TGFβ1 and extracts from Cervus korean TEMMINCK var. mantchuricus Swinhoe on acute and chronic arthritis in rats

Abstract

Aim of the Study

To elucidate the pharmacological activities of deer antler acupuncture and TGF1 on the acute and chronic phases of rheumatoid arthritis diseases.

Materials and Methods

Polyarthritis rats were administered with TGF1 and water extract of deer antler acupunture (DAA), prepared from the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe. TGF (0.1 to 2 g/animal) and DAA (5-100 g/kg animal) were initiated 1 day before an arthritogenic dose of streptococcal cell wall fragments to see the effects on the joint swelling and distortion during the acute phase and the chronic phase of the disease. Arthritic index suppression of rat arthritis model was examined by TGF and DAA administrations.

Results

TGF1 and DAA diminished the polyarthritis development in rats. TGF and DAA eliminated the joint swelling and distortion observed during the acute phase and the chronic phase of the disease. The TGF and DAA suppressed the arthritis progress when administration was begun after acute phase of arthritis.

Discussion

Consistent with the inhibition of inflammatory cell recruitment into the synovium, TGF1 and DAA reversed the leukocytosis associated with the chronic phase of the arthritis, respectively.

Introduction

Unossified horn or pilose antler cut from deer, which belong to the Cervidae, is generally termed “Nokyong”. Nokyong is one of the most famous Korean traditional medicines and has been considered to possess sexual-reinforcing and anti-aging actions. Nokyong or its extract has sometimes been compounded in recent Korean commercial restoratives, although little is yet known about the pharmacological effects or active ingredients. Water extract of deer antler aqua-acupuncture (DAA) has been widely used in the treatment of some immune-related diseases, especially rheumatoid arthritis (RA) and satisfactory results are obtained (Kim et al., 2003). Recently, we have reported that DAA treatment reduces the activation of proteolytic enzymes and free radicals, which are likely to be of equal potential importance as protein damaging agents in the pathogenesis of RA (Kim et al., 2008). DAA has been shown to reduce the chronic inflammation in adjuvant-induced arthritis rats (Kim et al., 2003, Kim et al., 2004a, Kim et al., 2004b). For example, when DAA was evaluated to assess the reductions in bone mass, strength, and turnover in adjuvant-induced arthritic rats, DAA at the doses of 10, 20, 50 and 100 μg/kg, administered by Shinsu (B23) acupuncture, significantly prevented the development of the chronic paw edema, indicating that DAA is able to prevent these reductions by modulating the bone turnover in this arthritis model (Kim et al., 2004b). To date several modern studies have shown only that extract of DAA can improve the syndrome occurring after whiplash injury and anemia in rabbits, and that polysaccharides and lysophosphatidylcholines are responsible for anti-ulcer and hypotensive actions, respectively (Wang et al., 1988a, Wang et al., 1988b). However, little is still known about the mode of action of DAA medication on RA in rodents.

For the nature of the compounds which could be responsible of the DAA activity, gangliosides, the sialic acid (NeuAc)-containing glycosphingolipids, have been suggested to be contained in the crude drug (Jhon et al., 1999). Five ganglioside fractions were isolated and purified from deer antler and GM3 and GD3 were present in the isolated ganglioside fractions. N-Acetyl GM3 (Neu5Ac), N-glycolyl GM3 (Neu5Gc), and N-acetyl GD3 (Neu5Ac) were also present in the antler. The major ceramide moiety was composed of C16:0 or C22:0 fatty acids along with either C18 sphingosine or C20 eicosasphingosine (Yang et al., 2004). Many other substances including free amino acids, polypeptides, trace elements, carbohydrates, hexosamines, mucopolysaccharides, uronic acids, sialic acids, prostaglandins, glycolipids, phospholipids, nucleic acids, hypoxanthine, cholesterol, cholest-5-ene-3,7α-diol, and cholesterol esters were also identified and claimed as the active components (Zhang et al., 1992, Zhao et al., 1992, Ivankina et al., 1993, Feng et al., 1995, Jhon et al., 1999, Yang et al., 2004).

The potent immunosuppressive effects of cytokine, transforming growth factor β (TGFβ) (Ristow, 1986), suggest that it may be valuable in the treatment of the immunological diseases. TGFβ has been shown to in vitro inhibit the proliferation of the immune-related cells including thymocytes, T- and B-lymphocytes, and hematopoietic progenitor cells, and production of immunoglobulins (Brandes et al., 1991). Previously, streptococcal cell wall (SCW)-induced chronic arthritis responses were modulated by TGFβ in rats (Cromatie et al., 1977).

In early research to find unidentified pharmacological effects of DAA, authors set out to investigate specific effects of DAA on arthritis animals by using an established arthritis mode (Kang et al., 2006). Then, we have evaluated DAA for its effectiveness on immune responses to type II collagen in the rat collagen-induced arthritis. The results showed that treatment with DAA, starting concurrently with either the initial or the booster immunization, can inhibit the onset and development of arthritis and the immune responses to collagen (Kang et al., 2006). Recently, we have observed the suppressive activity of DAA on the chronic RA model using SCW as an RA inducer, as reported for the TGFβ1. Therefore, to extend the pharmacological approach, the present study has been carried out to show that daily each administration of TGFβ1 and DAA reduce the acute and chronic phases of RA diseases with similar effects.