Involvement of inflammation in severe post-operative pain demonstrated by pre-surgical and post-surgical treatment with piroxicam and ketorolac.

OBJECTIVES: Post-operative pain is considered to involve inflammation caused by tissue injury. However, the mechanism and timing of the involvement of inflammation in the post-operative pain remain complicated because they can vary among different types of surgery. In this study a rat incision model was used to investigate how inflammation induced by cyclooxygenases (COXs) is involved in severe post-operative pain. METHODS: Longitudinal incision with a length of 1cm was made through skin and fascia on the right hind paw of rats, starting 0.5cm from the edge of the heel and extending towards the toes. Allodynia was evaluated using the von Frey hair test and its degree was recorded as the paw withdrawal threshold (PWT). Two non-steroidal anti-inflammatory drugs (NSAIDs), piroxicam and ketorolac, were given to rats after or before surgery, and the effects of the drugs on allodynia induced by the hind paw incision were examined. KEY FINDINGS: The PWT reduction reached a sub-maximal level at 3h, a maximal level at one day after the surgery and lasted for more than 8 days, with the parallel development of inflammation (characterized by cell infiltration and oedema). Treatment with morphine (1mg/kg, s.c.) at one day after the surgery showed a significant anti-allodynic effect. Treatment with either piroxicam (10mg/kg, p.o.) or ketorolac (10mg/kg, p.o.) at one day after the surgery did not exhibit significant anti-allodynic effect, whereas pre-surgical treatment with each drug showed significant anti-allodynic effects at 3h after surgery. CONCLUSIONS: These findings suggest the involvement of cyclooxygenases in evoking pain that occurs in the immediate post-operative period, and that an initial suppression of rapid inflammation by treatment with NSAIDs before major surgery plays an important role in the management of severe post-operative pain.

Anti-PDGF-B monoclonal antibody reduces liver fibrosis development.

AIM: To evaluate the usefulness of a platelet-derived growth factor (PDGF)-B specific monoclonal antibody (mAb) as a therapeutic agent to treat chronic liver fibrosis. METHODS: Liver fibrosis was induced in ICR mice by bile duct ligation (BDL) or BALB/c mice by weekly injection of concanavalin A (ConA) for 4 or 8 weeks. A mAb specific for mouse and human PDGF-B chain, AbyD3263, was generated, tested in vitro and administered twice a week throughout the experimental period. RESULTS: AbyD3263 showed neutralizing activity against mouse and human PDGF-B chain in cell-based assays, as measured in vitro by inhibition of phosphorylation of PDGF receptor ß and proliferation of hepatic stellate cells induced by PDGF-BB. The half life of AbyD3263 in mice exceeded 7 days and dosing of animals twice a week resulted in constant plasma levels of the mAb. Induction of liver fibrosis by BDL and ConA resulted in elevated levels of alanine aminotransferase (ALT) in plasma and hydroxyproline in the liver. Treatment with AbyD3263 did not modify ALT levels, but significantly reduced hydroxyproline content in the liver with a maximum reduction of 39% and 54% in the BDL and ConA models, respectively, compared to controls. Conclusios: Consistent with the notion that PDGF-BB plays an important role in the progression of liver fibrosis, AbyD3263 exhibits efficacy in pre-clinical disease models suggesting that pharmacological inhibition of PDGF-B chain may be a therapeutic approach to treat liver fibrosis.

Gene expression changes in dorsal root ganglion of rat experimental lumber disc herniation models.

STUDY DESIGN: Comprehensive overviews of gene expression changes in dorsal root ganglion (DRG) were obtained using microarrays in 2 rat models of experimental lumbar disc herniation (LDH). OBJECTIVE: To clarify the mechanisms of painful radiculopathy caused by LDH from the viewpoint of gene expression changes in DRG of 2 rat models of LDH. SUMMARY AND BACKGROUND DATA: Mechanical compression and chemical irritation are considered to be the 2 major causative factors of radiculopathy associated with LDH. Several basic studies have revealed histologic and functional changes in the nerve root and DRG induced by mechanical compression or application of nucleus pulposus. However, the effects of the 2 major factors have not been investigated in detail. METHODS.: The effects of mechanical and chemical factors were assessed in 2 models and in sham-operated rats. The mechanical compression model had a stainless steel rod inserted through a drill hole in the L5 lamina; the nucleus pulposus model had autologous nucleus pulposus placed in the drill hole, and only a drill hole was made in the L5 lamina of sham-operated rats. Samples from the left L5 DRG were harvested from the models with mechanical allodynia and from sham rats and analyzed using microarrays at 3 and 7 days after surgery. RESULTS: The gene expression profiles differed in the 2 models at 7 days, but were similar at 3 days after surgery. Expression of the growth factor gene, insulin-like growth factor 1, and of the cyclinD1, cell division cycle 2 homolog A, and cyclinA2 genes related to the cell cycle was significantly upregulated in the DRG of the mechanical compression group at 7 days after surgery. CONCLUSION: Mechanical and chemical factors caused altered gene expression in the DRG at 7 days after surgery, suggesting that the mechanisms of nerve injury induced by these factors differ. The upregulation of IGF-1 might be a key factor in painful radiculopathy induced by mechanical factors.

Discovery of (-)-6-[2-[4-(3-fluorophenyl)-4-hydroxy-1-piperidinyl]-1-hydroxyethyl]-3,4-dihydro-2(1H)-quinolinone--a potent NR2B-selective N-methyl D-aspartate (NMDA) antagonist for the treatment of pain.

(-)-6-[2-[4-(3-Fluorophenyl)-4-hydroxy-1-piperidinyl]-1-hydroxyethyl]-3,4-dihydro-2(1H)-quinolinone was identified as an orally active NR2B-subunit selective N-methyl-d-aspartate (NMDA) receptor antagonist. It has very high selectivity for NR2B subunits containing NMDA receptors versus the HERG-channel inhibition (therapeutic index=4200 vs NR2B binding IC(50)). This compound has improved pharmacokinetic properties compared to the prototype CP-101,606.

Upregulation of sodium-dependent vitamin C transporter 2 expression in adrenals increases norepinephrine production and aggravates hyperlipidemia in mice with streptozotocin-induced diabetes.

The hyperglycemia and hyperoxidation that characterize diabetes lead to reduced vitamin C (L-ascorbic acid, AA) levels in diabetic humans and animals. We examined the possibility that diabetes-induced low plasma AA levels impair AA distribution to various tissues and that these changes are closely related to the development of diabetic complications. AA levels were markedly decreased in the plasma and increased in the adrenals of mice with streptozotocin (STZ)-induced diabetes. Consistently with these results, in [1-(14)C]AA accumulation assays, the efficiency of [1-(14)C]AA accumulation was significantly higher in the adrenals (which had the greatest ability to accumulate [1-(14)C]AA) of diabetic mice than in those of controls. Expression of sodium-dependent vitamin C transporter (SVCT)-2, a transporter of AA, was upregulated in diabetic adrenals. Furthermore, increased AA incorporation into the diabetic adrenals by SVCT-2 led to increased plasma norepinephrine, triglyceride and free fatty acid levels in mice with STZ-induced diabetes. Therefore, oversupplementation with AA could be deleterious in diabetic patients, because overexpression of adrenal SVCT-2 in diabetes could lead to excessive AA uptake, thus enhancing norepinephrine production and exacerbating some diabetic complications. Interestingly, however, treatment with AA dose-dependently abolished the increased expression of adrenal SVCT-2 and normalized the abovementioned plasma parameters in diabetic mice. These results suggest SVCT-2-mediated increases in AA uptake by the adrenals followed by excessive production of plasma norepinephrine may play a pivotal role in the development of diabetic complications.

Pharmacological separation between peripheral and central functions of cyclooxygenase-2 with CIAA, a novel cyclooxygenase-2 inhibitor.

There are many reports concerning the physiological and pathological involvement of cyclooxygenase (COX)-2 in the central nervous system and peripheral tissue cells. Selective COX-2 inhibitors that mainly distribute peripherally have not been reported thus far. Therefore central and peripheral roles of COX-2 remain classified pharmacologically. In this study, in vivo pharmacological profiles of CIAA ([6-chloro-2-(4-chlorobenzoyl)-1H-indol-3-yl]acetic acid), a novel selective COX-2 inhibitor which distributes at higher concentrations in plasma than in brain, were compared with those of well-known selective COX-2 inhibitors, celecoxib and rofecoxib. Additionally, the possibility of pharmacological separation between peripheral and central actions of COX-2 with the inhibitors was investigated. CIAA selectively inhibited COX-2 activity compared with COX-1 in in vitro assays with rat whole blood. The compound exhibited lower brain penetration and higher plasma concentration (the brain/plasma concentration ratio was approximately 0.02) than celecoxib and rofecoxib after oral administration. Therefore, CIAA is mainly expected to act peripherally. Edema and prostaglandin E2 (PGE2) production in Carrageenan-injected rat paws, and pyrexia and PGE2 production in the brain in lipopolysaccharide (LPS)-injected rats were measured in in vivo experiments. CIAA exhibited lower ratios of anti-pyretic/anti-edematous activities and of inhibitory activities of PGE2 production in brain/paw than those of celecoxib and rofecoxib, and these ratios well-reflected brain/plasma concentration ratios. In conclusion, we discovered a novel selective COX-2 inhibitor, CIAA, which distributes at higher concentrations in plasma than in brain, which would make possible the pharmacological separation of the peripheral and central functions of COX-2.