The sensation of pain plays a critical role as an alert and as a protection system against tissue damage from mechanical, chemical, and thermal stimuli. Despite the protective role of pain, the severity of pain sensation is markedly attenuated by the endogenous pain inhibitory systems that predominantly originate at the brain stem. Both behavioral and in vivo extracellular recording studies have sought the loci producing analgesia and clarification of the anti-nociceptive actions. Among those loci, the main descending systems to the spinal dorsal horn are noradrenergic and serotonergic. Although, in vivo studies have provided basic knowledge of these systems, the precise synaptic mechanisms underlying the analgesic actions have not yet been elucidated until recently. The newly developed in vitro slice and in vivo patch-clamp recordings have disclosed the synaptic mechanisms of the noradrenergic and serotonergic effects at the level of spinal dorsal horn. This paper reviews the anti-nociceptive action of these systems, while particularly focusing on the electrophysiological aspects of the systems at the single neuron level in the spinal dorsal horn as well as their origins and responsible receptor subtypes.

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To evaluate the efficacy of three common antitussive Kampo formulas, eppikahangeto (EPP), bakumondoto (BAK), and shoseiryutogomakyokansekito (SGM), a new cough model of guinea pig was used, which could specifically induce a laryngeal cough by microinjection of citric acid solution into the larynx. Kampo extract was dissolved in water and the animals were given access ad libitum for 3 days, and then the number of coughs during 10 min was counted. EPP extract decreased the number of coughs dose-dependently (0.3% extract, −22.9 ± 6.6%, P<0.01; 1.0% extract, −32.4 ± 5.5%, P<0.01). BAK extract and SGM extract had no significant effect. Intraperitoneal injection of codeine (60 mg/kg) also decreased the number of coughs (−36.1 ± 9.1%, P<0.05). Furthermore, Ephedrae herba (EH) extract reduced the number of coughs (−18.3 ± 6.0%, P<0.05), but the extract of EPP without EH did not. These results suggest that EPP has an antitussive effect against laryngeally-induced cough in guinea pigs, and the crucial herbal medicine is EH.

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Nitric oxide (NO) induces apoptosis in various cells lines, while activation of the NO/cGMP signaling pathway prevents apoptosis induced by diverse stimuli, including NO. Here, we report the cytoprotective mechanisms of the NO/cGMP signaling pathway against NO-induced apoptosis in a mouse macrophage-like cell line, RAW264. Treatment with sodium nitroprusside (SNP), an NO donor, at a high-toxic concentration (4 mM) stimulated the N-terminal conformational change of Bax and its translocation to mitochondria followed by cytochrome c release and nuclear fragmentation in RAW264 cells. These changes of Bax were attenuated by pretreatment with SNP at a low-nontoxic concentration (100 μM) or dibutyryl cGMP (DBcGMP), a cell-permeable cGMP analogue. SB203580, a p38 mitogen-activated protein kinase (MAP kinase) inhibitor, blocked the effects of 4 mM SNP on Bax translocation and cell viability. Treatment with 4 mM SNP activated p38 MAP kinase and this effect was prevented by pretreatment with 100 μM SNP or DBcGMP. These findings suggest that the NO/cGMP signaling pathway inhibits NO-induced apoptosis of macrophages by suppressing the p38 MAP kinase activation, which results in N-terminal conformational change of Bax and its translocation to mitochondria.

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We investigated whether increased contractile responsiveness to epidermal growth factor (EGF) is associated with altered activation of mitogen-activated protein kinase (MAPK) in the aortic smooth muscle of deoxycorticosterone acetate (DOCA)-salt hypertensive rats. EGF induced contraction and MAPK activity in aortic smooth muscle strips, which were significantly increased in tissues from the DOCA-salt hypertensive rats compared with those from sham-operated rats. AG1478, PD98059, and LY294002, inhibitors of EGF receptor (EGFR) tyrosine kinase, MAPK/extracellular signal-regulated kinase (ERK) kinase, and phosphatidylinositol 3-kinase (PI3K), respectively, inhibited the contraction and the activity of ERK1/2 that were elevated by EGF. Y27632 and GF109203X, inhibitors of Rho kinase and protein kinase C, respectively, attenuated EGF-induced contraction, with no diminution of ERK1/2 activity. Although EGF also elevated the activity of EGFR tyrosine kinase in both sham-operated and DOCA-salt hypertensive rats, the expression and the magnitude of activation did not differ between strips. These results strongly suggest that EGF induces contraction by the activation of ERK1/2, which is regulated by the PI3K pathway in the aortic smooth muscle of DOCA-salt hypertensive rats.

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Overexpression of constitutively active (CA)-Gα₁₃ significantly increased the expression of interleukin (IL)-1β and IL-6 mRNAs and proteins in rat cardiac fibroblasts. IL-1β mRNA induction by CA-Gα₁₃ was suppressed by diphenyleneiodonium (DPI), an NADPH oxidase inhibitor, but not by BAPTA-AM, an intracellular Ca²⁺ chelator. In contrast, IL-6 mRNA induction by CA-Gα₁₃ was suppressed by BAPTA-AM but not by DPI. However, both IL-1β and IL-6 mRNA induction was suppressed by nuclear factor κB (NF-κB) inhibitors. The CA-Gα₁₃-induced NF-κB activation was suppressed by DPI and BAPTA-AM, but not C3 toxin and the Rho-kinase inhibitor Y27632. IL-6 mRNA induction by CA-Gα₁₃ was suppressed by SK&F96365 (1-[β-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole hydrochloride), an inhibitor of receptor-activated nonselective cation channels, and the expression of CA-Gα₁₃ increased basal Ca²⁺ influx. These results suggest that Gα₁₃ regulates IL-1β mRNA induction through the reactive oxygen species-NF-κB pathway, while it regulates IL-6 mRNA induction through the Ca²⁺-NF-κB pathway.

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Doxorubicin (DOX) is widely used to treat patients suffering from cancer, but the usage for patients is limited because of the dose-dependent cardiotoxicity. We hypothesized that DOX induces apoptosis through caspase activation in cardiomyocytes, and we examined this hypothesis using both rat primary cultured cardiomyocytes and rat hearts from an animal model. Cardiomyocytes were treated with DOX for 24 h. The activity of caspase-3 was significantly increased by DOX treatment. In rats with DOX injected intravenously once a week for 5 weeks, left ventricular fractional shortening evaluated by echocardiography was significantly decreased at age 14 weeks, 2 weeks after the end of DOX-administration. At 16 weeks of age, endothelin-1 mRNA and atrial natriuretic peptide mRNA were also significantly increased, likewise, and TUNEL positive cells were significantly increased in the ventricles of DOX-treated rats. The activity of caspase-3 in the ventricles was also significantly increased compared to that of untreated rats at 16 weeks. However, the activity of caspase-8 and the expression level of Fas-ligand mRNA were comparable with those of the untreated rats. In conclusion, DOX induces apoptosis through the activation of caspase-3, suggesting that apoptosis has an important role in the progression of cardiomyopathy due to DOX.

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We investigated whether long-term treatment with the angiotensin-converting enzyme (ACE) inhibitor imidapril or the calcium channel antagonist nifedipine increases systemic nitric oxide (NO) production in patients with essential hypertension. Twenty-nine patients with essential hypertension were randomly divided into two groups, and they were treated with either imidapril or nifedipine once daily p.o. for 4 weeks. Long-term treatment with imidapril significantly decreased blood pressure and increased plasma NOx concentration. Long-term treatment with nifedipine also caused a comparable extent of significant decrease in blood pressure, but failed to alter plasma NOx levels. The imidapril treatment significantly inhibited serum ACE activity and increased plasma bradykinin concentration. Furthermore, the extent of inhibition of serum ACE activity and the extent of increase in plasma bradykinin concentration in response to the imidapril treatment were both significantly correlated with the extent of increase in plasma NOx concentration. In contrast, no such changes were noted after the nifedipine treatment. These results provide the first evidence that long-term treatment with imidapril enhances plasma NOx concentration in patients with essential hypertension. This effect does not seem to be due to the decrease in blood pressure. The increase in bradykinin concentration may be involved in the enhancing effect of the ACE inhibitor on NOx production in vivo.

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It has been known that oxidative stress plays an important role in the development of diabetic nephropathy (DN). The antioxidative effects of Astragalus saponin I (AS I) were studied in vitro and in vivo. In the presence of high glucose and H₂O₂, the total antioxidative capability, catalase, reduced glutathione, and superoxide dismutase level of rat mesangial cells were significantly decreased, and transforming growth factor β1 (TGF-β1) mRNA level, collagen IV, and laminin level were significantly increased. When compared with those in the high glucose group, these 4 indexes of cells incubated in 2.0 and/or 20 μmol/L of AS I were significantly enhanced, and levels of TGF-β1 mRNA, collagen IV and laminin were statistically decreased. By flowcytomery, percentages of S phase of cells incubated in high glucose and H₂O₂ were lowered, while those in AS I were increased. Furthermore, the physical behaviors of rats treated with 12 mg/kg of AS I restored with vigor and weight gaining, while the level of HbAlC was significantly reduced. Thus, AS I has antioxidative effects and is a potential compound worth further study because it may prevent the development of DN.

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Intracellular ADP-ribose is an activator of TRPM2, which is a Ca²⁺-permeable channel and mediates H₂O₂-induced cell death, in the TRPM2-expressing rat β-cell line RIN-5F. We examined the effect of extracellular-added ADP-ribose on intracellular Ca²⁺ concentration in RIN-5F cells. ADP-ribose induced Ca²⁺ release from the thapsigargin-sensitive Ca²⁺ store, but not Ca²⁺ entry across the plasma membrane. A phospholipase C (PLC) inhibitor and a non-specific IP₃ receptor inhibitor blocked its Ca²⁺ release. H₂O₂-induced Ca²⁺ entry through TRPM2 was not affected by extracellular ADP-ribose. These findings suggest that extracellular-added ADP-ribose induces Ca²⁺ release through the PLC-IP₃ pathway and does not act as a TRPM2 activator.

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Wrong:Proteosome
Right:Proteasome

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