Neuropeptide Organization of the Nociceptive and Antinociceptive Brain Systems in the Cat

Using radioimmune techniques, we studied in detail the concentrations of -endorphin (-En), met- and leu-enkephalins (mE and lE, respectively), and substance P (SP) in a number of structures of the brainstem and forebrain of the cat. According to the proposed concept, these structures comprise the noci- and antinociceptive brain systems (NS and ANS). The above indices were measured in intact animals and in animals after nociceptive electrocutaneous stimulation (NECS) of the limb, stimulation of the ventrolateral zone of the midbrain central gray (vl SGC, a nociceptive midbrain structure), stimulation of the dorsolateral part of the above region and dorsal raphe nucleus (dl SGC and Rd, antinociceptive midbrain structures), and after combined stimulations (NECS preceded by conditioning stimulation of one of the above midbrain zones). We found that in the norm maximum SP concentrations were observed in the NS structures, while those of -En and mE were the highest in the hypothalamic nuclei belonging to the ANS and in its midbrain centers (dl SGC and Rd). Nociceptive ECS, stimulations of the studied midbrain zones, and combinations of these stimulations could result in specific and, in some cases, very significant (by an order of magnitude and more) shifts in the concentrations of the mentioned neuropeptides in the studied set of the central structures. After NECS and its combination with vl SGC stimulation, SP concentrations in the NS structures considerably increased, while -En and mE concentrations in the ANS components dropped. Stimulations of the dl SGC and Rd were accompanied by increases in the mE and -En levels and simultaneous drops in the SP concentrations in the ANS components; reciprocate shifts were observed within the NS. Changes in the lE level, which were related to the influences used, were less specific and mostly appeared as increases in this index in the structures of both the NS and ANS. Combinations of NECS with conditioning stimulations of the vl SGC, dl SGC, or Rd demonstrated that the latter exert significant modulatory effects on the NECS-induced shifts in the concentrations of the studied neuropeptides. Considering the obtained data, a hypothetical scheme of neuropeptide organization of the cerebral NS and ANS has been proposed. In the examined brain structures, there are neuronal populations belonging to the two main neurochemical systems. One of them is SP-ergic, while another consists of mE- and -En-ergic neurons; these systems are in antagonistic relations. Changes in the levels of mE and -En always induce the attended opposite shifts in the SP levels, and vice versa. The lE-ergic neuronal populations, which co-exist with the above neurochemical systems, are relatively nonspecifically activated by either (noci- and antinociceptive) drives, but, according to the pattern of its responses, the lE-ergic system is closer to the SP-ergic one. It is supposed that pain signals, when coming to the vl SGC, activate SP- and lE-ergic neuronal populations; later on, the posterior and lateral hypothalamic nuclei and preoptic region are involved in the transmission of the above signals. When released by the corresponding neuronal populations in the vl SGC, lE activates the key ANS structures (dl SGC and Rd), and the latter, in turn, activate other components of this system, which form its ascending compartment (ventromedial, dorsomedial, and paraventricular hypothalamic nuclei, septum, basolateral amygdala, hippocampal fields 3 and 4, and cingular cortex). In the ANS,-En and mE function as transmitters.     

Changes in Glycosyldiacylglycerols during Germination of Black Mung Beans (Matpe)

The physico-chemical characteristics of purified arginine kinases from prawn and swimming crab were examined. The molecular weights of prawn and swimming crab enzymes were 40,500 and 40,000, respectively. Amino acid analysis indicated that there were some differences in the contents of proline, glycine, methionine, and lysine. The other amino acid compositions of these enzymes resembled each other.

Both enzymes were stable up to 20°C when they were treated for 10 min at various temperature levels. The enzymes lost their activities at temperatures higher than 25°C. They were more stable at pH 8.0 than pH 7.0. The optimum temperature for the enzyme of prawn was about 42°C and that for swimming crab was about 40°C. The pH optima for the activity of arginine kinase of prawn in the forward and in the reverse reactions were found to be 9.0 and 6.1, respectively. For the swimming crab, the similar optimum pHs at 9.2 in the forward reaction and 5.8 in the reverse reaction were observed. Both enzymes were activated most strongly with Mg^{2 +} and Mn^{2 +} followed by Ca^{2 +}, Co^{2 +}, and Fe^{2 +}. The enzymes were not activated by Sr^{2 +}, Cu^{2 +}, or Zn^{2 +}.

The optimum molar ratio of Mg^{2 +}: ATP in the forward reaction of prawn and swimming crab was found to be 1:1, and the molar ratio of Mg^{2 +} : ADP in the reverse reaction was 4:1 in both cases. Kinetic studies indicated that dissociation constants were rather different. In the prawn, dissociation constants for arginine, ATP, AP, and ADP were 0.19,0.31, 0.67, and 0.29 mM, respectively, but in the swimming crab, they were 0.10, 0.18, 0.22, and 0.11 mM, respectively.     

Combination of pharmacological analgesics and microwave irradiation of an acupuncture point for suppression of visceral pain in mice: Role of the opioid and serotonergic cerebral systems

We studied suppression of pain-related reactions induced in mice by i.p. injection of 0.08 ml of a 2% solution of acetic acid using pharmacological analgesics (analgin and tramadol) combined with low-intensity microwave irradiation of an acupuncture point (AP) E-36 (frequency 30 to 300 GHz and power rate density 3·10⁻⁹ W/cm²). The respective effects were also observed under conditions of suppression of the functions of opioid and serotonergic cerebral systems using injections of, respectively, naloxone and DL-p-chlorophenylalanine (p-CPA). We found that antinociceptive effects provided by analgesics used in a 50% mean single dose in the combination with microwave irradiation of the AP were significantly more intense than those induced by isolated injection of analgesics used in both 50% and full mean single doses and isolated microwave irradiation of the AP E-36. After injections of naloxone, analgesic effects caused by the combined action of analgin and microwave irradiation of the AP were considerably smaller. At the same time, after injection of p-CPA, analgesic effects, provided by the combination of injection of pharmacological agents and microwave irradiation of the AP, weakened in the case of use of both analgesics. This was manifested in a significant increase in the total duration of pain-related behavioral reactions. Therefore, the studied analgesic effects observed in the examined animal groups are realized due to the involvement of the opioid and serotonergic cerebral systems. Neirofiziologiya/Neurophysiology, Vol. 39, No. 6, pp. 468–477, November–December, 2007.     

Stereoselectivity in central analgesic action of tocainide and its analogs

The antiarrhythmic drug tocainide (5a) and some related chiral α-amino and α-imino anilides (5b–e) were synthesized in optically active form. The antinociceptive effects of the different stereoisomers of these compounds were examined and it was found that the analgesic effect of tocainide is due only to its (?)-(R)-enantiomer. Benzyl replacement for methyl group at the stereogenic centre of tocainide causes loss of activity while both enantiomers of the αiminoxilidide 5e and of the strictly related tocainide analog 5d produce an analgesic effect without any stereoselectivity. Pharmacological tests and [³H] quinuclidinyl benzilate ([³H]QNB) binding assay, taken together, seem to show that the antinociceptive effect of (?)-(R)-tocainide, like the analgesia induced by lidocaine, procaine, and mexiletine, is due to a central presynaptic cholinergic mechanism of action. © 1993 Wiley-Liss, Inc.     

Synthesis of some imidazolyl-thioacetyl-pyrazolinone derivatives and their antinociceptive and anticancer activities

In this study, some 4-(1,5-diarylimidazol-2-yl)thioacetyl-1-phenyl-2,3-dimethyl-3-pyrazoline-5-one derivatives were prepared by reacting 4-(2-chloroacetyl)-1-phenyl-2,3-dimethyl-3-pyrazoline-5-one and 2-mercapto-1, 5-diarylimidazole derivatives. The antinociceptive and anticancer activities of the compounds obtained were investigated. It was observed that some of the compounds, 2a, 2d, 2g, and 2j, showed remarkable antinociceptive activity, and one of the compounds, 2i, showed weak anticancer activity.     

Microwave-assisted extraction and pharmacological evaluation of polysaccharides from Posidonia oceanica

Microwave-assisted extraction was employed for the isolation of polysaccharides from Posidonia oceanica (PPO). The extracting parameters were optimized adopting response surface methodology. The highest polysaccharide yield (2.55 ± 0.09%), which is in concordance with the predicted value (2.76%), was obtained under the following conditions: extraction time 60 s, liquid–solid ratio of 50:1 (mL/g) and power of 800 W. This polysaccharide, with molecular weight of 524 KDa, characterized by gas chromatography–mass spectrometry showed that PPO was mainly composed of galactose, glucose, and arabinose with molar percentages 25.38, 24.37, and 21.64%, respectively. The pharmacological evaluation of PPO using animal models at the dose of 100 mg/kg indicated a significant anti-inflammatory activity with a percentage of inhibition of edema of 54.65% and a significant antinociceptive activity with 78.91% inhibition of writhing for peripheral analgesic activity and an increase in the hot plate reaction time for central analgesic activity.     

臭灵丹水提取物的急性毒性及镇痛作用的实验研究

采用上下移动法,醋酸扭体法、热板法、福尔马林致痛试验对臭灵丹水提取物的急性毒性及镇痛作用进行了研究。结果显示：臭灵丹水提取物LD50（腹腔注射）为1．19g／kg;臭灵丹水提取物明显抑制醋酸所致的小鼠扭体数,显著减少福尔马林致痛试验后期小鼠舔足行为;而对热板法所致疼痛无明显作用。表明臭灵丹水提取物具有一定的外周镇痛作用。     

Synthesis and antinociceptive-antimicrobial activities of some new amide derivatives of 3,5-di/-and 1,3,5-trimethylpyrazoles

Some N-(3,5-di-/1,3,5-trimethylpyrazole-4-yl)-4-substitutedbenzamide derivatives were prepared as possible antiociceptive-antimicrobial agents. New amide derivatives (3–12) were synthesized by reacting 4-amino-3,5-di and 1,3,5-trimethylpyrazoles with 4-substitutedbenzoyl chlorides. Hotplate and tail-immersion tests were used for the determination of the antinociceptive activity. Morphine, was used as a standard test drug. All compounds were administered at a dose of 100 mg/kg ip and some of them had significant antinociceptive activity in both tests. Compound 10 (N-(1,3,5-trimethylpyrazole-4-yl)-4-bromobenzamide), was the most active one in both tests among the compounds. The antinociceptive activity of the compounds 10, 11 (N-(1,3,5-trimethylpyrazole-4-yl)-4-chlorobenzamide), and 12 (N-(1,3,5-trimethylpyrazole-4-yl)-4-fluorobenzamide), started at 30 minutes and continued up to 150 minutes in the hotplate test. Also compounds were tested for their in vitro antimicrobial activity, but exhibited weak antibacterial activity.     

Roles of different neurochemical systems in mechanisms underlying the antinociceptive effect of extrahigh-frequency electromagnetic radiation

We studied the effect of low-intensity extrahigh-frequency (EHF) electromagnetic radiation (EMR) on the duration of a pain behavioral reaction in rats under conditions of experimental induction of tonic pain (formalin test). The antinociceptive effect of EHF irradiation was modulated by suppression of the activity of a few neurochemical systems resulting from the blockade of receptors of opioid peptides, α-and β-adrenoreceptors, receptors of dopamine and melatonin, as well as from inhibition of serotonin synthesis. We demonstrated that all the respective neurochemical systems are to a certain extent involved in the mechanisms underlying the analgesic action of EHF EMR. Within an early phase of pain stress, functioning of the opioidergic and noradrenergic systems and the effects of melatonin play leading roles, while the activity of the serotonergic system plays such a role within the second (tonic) phase. Neirofiziologiya/Neurophysiology, Vol. 39, No. 2, pp. 165–173, March–April, 2007.     

大黄总蒽醌对小鼠扭体模型的镇痛作用

目的:观察大黄总蒽醌对小鼠扭体模型的镇痛作用。方法:60只小鼠随机分为对照组、大黄总蒽醌组、地塞米松组(n=20),给予各组小鼠灌胃相应药物,1次/d,给药5天。于第5天给药后30min用醋酸扭体法测定各组扭体次数。结果:大黄总蒽醌具有较强的镇痛活性,而且呈现出剂量依赖性关系,最大的剂量600mg/kg,可达到55.56%的镇痛效果,大黄总蒽醌组和地塞米松组小鼠的扭体次数少于对照组(P<0.05,P<0.01)。结论:大黄总蒽醌对冰醋酸所致小鼠扭体模型有一定的镇痛作用。