Oral administration of type II collagen suppresses pro-inflammatory mediator production by synoviocytes in rats with adjuvant arthritis.

The objective of this study was to investigate the effect of the oral administration of type II collagen (CII) on pro-inflammatory mediator production by synoviocytes in rats with adjuvant arthritis (AA). Sprague-Dawley rats were fed with bovine CII either before immunization with Complete Freund's adjuvant (CFA) or after initiation of arthritis. Hind paw secondary swelling was measured and synoviocytes were harvested. Sera from portal vein of oral tolerized rats were collected and in vitro synoviocytes culture or synoviocytes-Peyer's Patches (PP) cells coculture system were developed. Interleukin (IL)-1 activity was measured by a mouse thymocyte activation assayed by MTT dye reduction and tumour necrosis factor (TNF) activity was measured by an L929 cytotoxicity bioassay. Nitric oxide (NO) and malondialdehyde (MDA) levels were measured by biochemical methods. We found that feeding with CII (5, 50 and 500 micro g/kg) for 7 days before immunization significantly suppressed hind paw secondary swelling measured at day 16, 20, 24 and 28 (all P < 0.01) and pro-inflammatory mediator (IL-1, TNF, NO and MDA) production by synoviocytes (all P < 0.01) in rats with AA. Feeding with CII (5, 50 and 500 micro g/kg) for 7 days after initiation of arthritis had a similar effect. CII (1, 10, 100 micro g/ml) had no effect on IL-1 and TNF production by synoviocytes in vitro, but CII 10 micro g/ml suppressed IL-1 and TNF production by synoviocytes-PP cells coculture system (P < 0.01), which was antagonized by anti-TGF-beta antibody (10 micro g/ml) (P < 0.01). Portal serum (1 : 10) from oral tolerized rats suppressed IL-1 and TNF production by synoviocytes (P < 0.01), which was also antagonized by anti-TGF-beta antibody (10 micro g/ml) (P < 0.01). We conclude that oral administration of CII had prophylactic and therapeutic effects on AA and over-production of IL-1, TNF, NO and MDA by synoviocytes was suppressed. Bystander active suppression may be the main mechanism of oral CII in the suppression of synoviocyte function.

Antinociceptive effect of astragalosides and its mechanism of action.

AIM: To study the effect and mechanism of astragalosides (AST) related to the antinociceptive activity. METHODS: The standardized formalin test was performed to induce the direct stimulation of nociceptors followed by inflammatory process in the Kunming strain mice. The involvement of opioid and nitric oxide was studied by subcutaneous injection of morphine with/without naloxone 30 min before formalin test, or peritoneal injection of L-arginine with/without L-NAME 20 min before formalin. RESULTS: AST 20, 40, and 80 mg/kg significantly lowered pain score of the second phase of formalin response as compared with control group (P<0.01). The maximum analgesic effect of AST 40 mg/kg was found at 4 h after the administration of AST (34.4 % inhibition at the second phase). Injection of morphine 5 mg/kg significantly inhibited pain response of both phases (P<0.01) and this was reversed by naloxone 2 mg/kg (P<0.01). However, naloxone did not alter the effect of AST on the second phase. Antinociceptive effect of AST 40 mg/kg was partially blocked by L-arginine 400 or 800 mg/kg (P<0.01). CONCLUSION: AST has an antinociceptive effect on formalin test in mice that is not mediated by the endogenous opioid system but related to its inhibitory effect on the production of NO.

Facilitation of AVP(4-8) on gene expression of BDNF and NGF in rat brain.

In situ hybridization and Northern blot assay were used to evaluate the effects of exogenous AVP(4-8) on the transcription of mRNAs for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3) in the adult rat brain. NGF and BDNF expression was found to be significantly enhanced by AVP(4-8) administration in the cerebral cortex and hippocampus, but NT-3 expression was not changed. In the same conditions, behavior-active arginine-vasopressin (AVP) showed a small effect and its behavior-inactive homologue, oxytocin did not. Our results suggest that selective regulation of neurotrophin gene expression by the peptides may be responsible for its memory-enhancing function.

Enhancement of NGF gene expression in rat brain by the memory-enhancing peptide AVP(4-8).

Northern blot analysis of nerve growth factor (NGF) was used to evaluate the effect of exogenous AVP(4-8) on the transcription of NGF gene in rat brain. NGF expression was found to be significantly enhanced by exogenous AVP(4-8) in the hippocampus as well as in the cerebral cortex in a time period of 12 h. This effect was inhibited by an antagonist to AVP(4-8). In addition, gel mobility shift assay was also used to observe the in vitro expression of c-fos gene in rat hippocampal slices. Our results suggest that NGF gene is one of the target genes responsible for memory-enhancing responses induced by AVP(4-8) and that the enhancement of NGF gene expression may share the signaling pathway mediated by AVP(4-8) receptor and c-fos gene expression.

NLPR, an agonist of AVP4-8, increases NGF gene expression in memory-impaired rat brain.

Oral administration of the tetrapeptide Asn-Leu-Pro-Arg (NLPR) to memory-impaired rats results in improved acquisition and maintenance of behavioural response and also facilitates nerve growth factor (NGF) expression in the brain. It is suggested that NLPR can ameliorate memory disability by promoting NGF gene expression, so implying that NLPR is a potential drug candidate for curing memory impairment.