Association of terpinolene and diclofenac presents antinociceptive and anti-inflammatory synergistic effects in a model of chronic inflammation

Pharmacological treatment of inflammatory pain is usually done by administration of non-steroidal anti-inflammatory drugs (NSAIDs). These drugs present high efficacy, although side effects are common, especially gastrointestinal lesions. One of the pharmacological strategies to minimize such effects is the combination of drugs and natural products with synergistic analgesic effect. The monoterpene terpinolene (TPL) is a chemical constituent of essential oils present in many plant species, which have pharmacological activities, such as analgesic and anti-inflammatory. The association of ineffective doses of TPL and diclofenac (DCF) (3.125 and 1.25 mg/kg po, respectively) presented antinociceptive and anti-inflammatory effects in the acute (0, 1, 2, 3, 4, 5 and 6 h, after treatment) and chronic (10 days) inflammatory hyperalgesia induced by Freund's complete adjuvant (CFA) in the right hind paw of female Wistar rats (170-230 g, n=6-8). The mechanical hyperalgesia was assessed by the Randall Selitto paw pressure test, which determines the paw withdrawal thresholds. The development of edema was quantified by measuring the volume of the hind paw by plethismography. The TPL/DCF association reduced neutrophils, macrophages and lymphocytes in the histological analysis of the paw, following a standard staining protocol with hematoxylin and eosin and the counts were performed with the aid of optical microscopy after chronic oral administration of these drugs. Moreover, the TPL/DCF association did not induce macroscopic gastric lesions. A possible mechanism of action of the analgesic effect is the involvement of 5-HT2A serotonin receptors, because ketanserin completely reversed the antinociceptive effect of the TPL/DCF association. These results suggest that the TPL/DCF association had a synergistic anti-inflammatory and analgesic effect without causing apparent gastric injury, and that the serotonergic system may be involved in the antinociceptive effect of this association.

Migração espermática em suínos após inseminação artificial intrauterina profunda / Sperm migration in pigs after deep intrauterine artificial insemination

A inseminação artificial intrauterina profunda (IIP) é de grande importância para a indústria suinícola, em função do maior número de doses produzidas por reprodutores de alto mérito genético e da possibilidade da utilização de biotecnologias, como sêmen sexado e/ou congelado. Entretanto, necessita-se compreender com maior propriedade os mecanismos pelos quais os espermatozoides colonizam as tubas uterinas. Assim sendo, pretende-se com o presente experimento avaliar a existência ou não de migração intraperitoneal de espermatozoides inseminados profundamente em um dos cornos uterinos, mediante a obtenção de oócitos fertilizados no corno contralateral à inseminação e seccionado na base, na junção com o corpo do útero. Quatorze fêmeas pluríparas foram divididas em dois grupos experimentais, sendo que em um deles as fêmeas foram submetidas à secção da base de um dos cornos uterinos (Grupo Operado, n = 7), enquanto as do Grupo Controle (n = 7) não foram submetidas a nenhuma intervenção cirúrgica. Ambos os grupos foram submetidos à IIP, sendo as fêmeas abatidas 5±1,2 dias após a última inseminação. Os sistemas genitais das fêmeas foram coletados, dissecados e o número de corpos lúteos contados em ambos os ovários. A recuperação dos embriões foi feita por meio de lavagem das tubas e cornos uterinos com solução de PBS (Phosphate Buffered Saline), após o que se avaliou os fluidos coletados em lupa para a identificação de embriões. Em ambos os grupos experimentais, foram encontrados embriões nos segmentos do sistema genital de ambos os lados. Apenas uma fêmea apresentou embriões nos segmentos em somente um dos lados no grupo operado. Diante dos resultados aqui observados, concluiu-se que a migração espermática no suíno pode ocorrer tanto por via retrógrada pelo útero quanto por migração intraperitoneal. Estes achados certamente contribuirão para aumentar a eficiência da técnica de IIP, sendo de grande valia para o aprimoramento da indústria suinícola...

Deep intrauterine insemination (DUI) is of great importance for the swine industry as it can increase the efficiency in the use of boars of high genetic merit, and facilitate the use of biotechnologies such as frozen and sexed semen. However, a better understanding of the mechanisms by which the sperm colonize the uterine tubes is essential. The aim of the present study was to investigate the existence of intrauterine sperm migration after DUI in one uterine horn, through the fertilization of oocytes in the contra lateral uterine horn. Fourteen multiparous sows were divided into two experimental groups: Operated (n = 7), where females had a segment close to the base of the uterine horn surgically removed, and Control (n = 7), females with intact uterus. Both groups were inseminated through DUI and slaughtered 5±1.2 days after the last insemination. The reproductive tracts collected were dissected and the number of corpora lutea counted in both ovaries. Embryo recovery was performed though flushings of uterine tubes and horns with Phosphate Buffered Saline solution and further examination under a dissecting microscope. Embryos were found in the uterine horns of both sides of the reproductive tract in both experimental groups. In the operated group, just one female had embryos in only one side of the reproductive tract. The results presented herein suggest that sperm migration in pigs may occur both in a retrograde way through the uterus and by intraperitoneal migration. These findings will certainly contribute to increase the efficiency of the DUP technique, which is of great importance for the improvement of the swine industry...

Antinociceptive effect on mice of the hydroalcoholic fraction and (-) epicatechin obtained from Combretum leprosum Mart & Eich

Previous studies on Combretum leprosum, a tree growing in the Northeastern states of Brazil, have shown antinociceptive effects of the ethanol extract of its leaves and bark, but studies examining its constituents are rare. The objective of this study was to evaluate the antinociceptive effect of the hydroalcoholic fraction (HF) of one of its constituents, the flavonoid (-) epicatechin (EPI), administered orally to mice (20-30 g) in models of chemical nociception, and the possible mechanisms involved. Different doses of HF (62.5 to 500 mg/kg) and EPI (12.5 to 50 mg/kg) were evaluated in models of abdominal writhing, glutamate, capsaicin, and formalin in animals pretreated with different antagonists: naloxone, ondansetron, yohimbine, ketanserin, pindolol, atropine, and caffeine in the abdominal writhing test. To determine the role of nitric oxide, the animals were pretreated with L-arginine (600 mg/kg, ip) in the glutamate test. The HF was effective (P < 0.05) in all protocols at different doses and EPI was effective in the abdominal writhing, capsaicin and glutamate tests (P < 0.05) at doses of 25 and 50 mg/kg. However, in the formalin test it was only effective in the second phase at a dose of 25 mg/kg. The antinociceptive effect of HF was inhibited when HF was associated with yohimbine (0.15 mg/kg), ketanserine (0.03 mg/kg), and L-arginine (600 mg/kg), but not with the other antagonists. HF and EPI were effective in models of chemical nociception, with the suggested participation of the adrenergic, serotonergic and nitrergic systems in the antinociceptive effect of HF.

Efeitos de diferentes fontes de energia sobre taxa ovulatória, fertilidade e sobrevivência embrionária em marrãs cíclicas / Effects of different energy sources on ovulation rate, pregnancy rate and embryo survival in cyclic gilts

Cinqüenta e quatro marrãs cíclicas, uniformizadas quanto à linhagem, família, ganho de peso, espessura de toucinho, peso, precocidade sexual, número de cios e escore clínico, foram alocadas em dois grupos experimentais com dietas isocalóricas, isoprotéicas e isolisínicas. Duas fontes de energia foram testadas: amido de milho (T1) e óleo de soja (T2). Sincronizou-se o segundo estro com allyl-trenbolone, para inseminação no terceiro estro. Foi realizada cateterização não cirúrgica em 21 marrãs, submetidas a dois ciclos de coleta para dosagem de glicose e insulina, aos 14 e 21 dias do ciclo. Todas as marrãs foram abatidas aos 28,6 dias de gestação média, para análises biométricas do trato reprodutivo. Marrãs do T1 apresentaram maior taxa ovulatória em relação às do T2 (16,52 vs 14,70, P<0,01). Não houve diferença entre os tratamentos nas taxas de prenhez e sobrevivência embrionária. É possível alterar a eficiência reprodutiva por intermédio de manipulação dietética, mesmo em marrãs em estado anabólico. O uso do amido de milho na fase pré-cobertua melhorou a eficiência reprodutiva dos animais avaliados.

Fifty four cyclic gilts were randomly selected and uniformized according to genetic background, litter of origin, weight gain, backfat, number of cycles and clinical score. Gilts were alloted to one of two groups fed isocaloric, isoproteic and isolysinic diets. Two energy sources were tested: corn starch (T1) and soybean oil (T2). Second estrus was synchronized with oral allyl-trenbolone, so that insemination was carried out at third estrus. Indweeling catheters were implanted by non-surgical technic in 21 gilts, which were submitted to consecutive blood samplings for glicose and insulin determination. Timing of ovulation was estimated by means of ultrasonography. All gilts were slaughtered at an average gestation lenght of 28.6 days. Starch-fed gilts (T1) showed higher ovulation rates than T2 gilts (16.52 vs 14.70; P<0.01). There was no effect of treatments on pregnancy rate and embryo survival. Results indicate it is possible to manipulate reproductive efficiency through diet even in anabolic experimental models like cyclic gilts. Feeding starch as main energy source during pre-mating flushing phase improved reproductive efficiency of cyclic gilts.

Chronic intrathecal cannulation enhances nociceptive responses in rats

The influence of a chronically implanted spinal cannula on the nociceptive response induced by mechanical, chemical or thermal stimuli was evaluated. The hyperalgesia in response to mechanical stimulation induced by carrageenin or prostaglandin E2 (PGE2) was significantly increased in cannulated (Cn) rats, compared with naive (Nv) or sham-operated (Sh) rats. Only Cn animals presented an enhanced nociceptive response in the first phase of the formalin test when low doses were used (0.3 and 1 percent). The withdrawal latency to thermal stimulation of a paw inflamed by carrageenin was significantly reduced in Cn rats but not in Nv or Sh rats. In contrast to Nv and Sh rats, injection in Cn animals of a standard non-steroid anti-inflammatory drug, indomethacin, either intraperitoneally or into the spinal cord via an implanted cannula or by direct puncture of the intrathecal space significantly blocked the intensity of the hyperalgesia induced by PGE2. Cannulated animals treated with indomethacin also showed a significant inhibition of second phase formalin-induced paw flinches. Histopathological analysis of the spinal cord showed an increased frequency of mononuclear inflammatory cells in the Cn groups. Thus, the presence of a chronically implanted cannula seems to cause nociceptive spinal sensitization to mechanical, chemical and thermal stimulation, which can be blocked by indomethacin, thus suggesting that it may result from the spinal release of prostaglandins due to an ongoing mild inflammation

Evaluation of the purified fraction of Wilbrandia (c. f. ) verticillata for antitumour activity

Cucurbatacins are known to produce cytotoxic and anticancer activities. Two novel norcucurbitacin glucosides (Wvl and Wv2) have recently been isolated from a purified fraction obtained from the rhizome of Wilbrandia verticillata. The present study evaluates the cytotoxic and anti-tumour activities of the norcucurbitacins. We have found a regular cytotoxicity in KB cells (Cy50 = 12µg/ml) as well as a significant inhibition in the Walker 256 carcinosarcoma growth (approximately 75%).

Inhibition of formaldehyde-induced arthritis by a purified fraction prepared from Wilbrandia (cf) verticillata which contains novel norcucurbitacin glucosides

The anti-inflammatory activity of a purified fraction of the rhizome of Wilbrandia (cf) verticillata, which contains two novel norcucurbitacin glucosides, is reported. The increase of vascular permeability induced by acetic acid in mice (N = 5) was inhibited 69% and 90% by 50 and 100 mg/kg of the purified fraction, po (P<0.01). Acetylsalicylic acid (200 mg/kg), po, inhibited the response by 62% under the same conditions (P<0.05). The purified fraction (100 mg/kg, po) also significantly inhibited paw swelling in the rat formaldehyde-induced arthritis model on 8 of 10 days and reduced the swelling by 63% on day 10. Dexamethasone (1 mg/kg, ip) was more effective than the extract under the same conditions. These data partially characterize the anti-inflammatory activity of the purified fraction from this plant which is used in Brazilian folk medicine for the treatment of arthritis and related disorders