Inhibition of the CXCR3-mediated pathway suppresses ultraviolet B-induced pigmentation and erythema in skin.

BACKGROUND: Skin pigmentation by ultraviolet (UV) B radiation is caused in part by inflammation mediated by chemokines and cytokines secreted by keratinocytes in the irradiated area. However, such inflammatory processes have not been well documented. OBJECTIVES: To elucidate the inflammation processes caused by UVB irradiation using skin-lightening agents that suppress melanin synthesis after UVB irradiation. METHODS: Utilizing a three-dimensional (3D) skin model, agents that suppressed formation of sunburn cells (SBC) after UVB irradiation were screened. Molecules whose expression was upregulated by UVB irradiation and attenuated by pretreatment with the agent were then screened by gene microarray to explore the mechanism of UVB irradiation. Messenger RNA expression of the molecules identified to be responsible for melanin biosynthesis was knocked down with a Tet-off shRNA lentivirus construct to confirm the involvement of the molecule in the pigmentation pathway following UVB irradiation. RESULTS: Paeonia suffruticosa Andrews (PSA) pretreatment suppressed SBC formation in the 3D skin model, and erythema formation and pigmentation in volunteers exposed to UVB irradiation. Comprehensive gene analysis after UVB irradiation showed upregulation of CXCR3 and its ligands, CXCL9/monokine induced by interferon (IFN)-γ (MIG), CXCL10/10-kDa IFN-γ-induced protein (IP-10) and CXCL11/inducible T-cell α-chemoattractant (I-TAC). Upregulation of these genes was partially suppressed by PSA pretreatment. Melanin biosynthesis increased upon stimulation of CXCR3 ligands (MIG, IP-10 or I-TAC) and decreased following CXCR3 downregulation by shRNA knockdown. CONCLUSIONS: UVB irradiation activates CXCR3-mediated signalling that leads to melanin synthesis. PSA pretreatment shows a lightening effect partly by attenuating CXCR3-mediated signalling at the transcriptional level.

Eicosanoid-induced Ca2+ release and sustained contraction in Ca(2+)-free media are mediated by different signal transduction pathways in rat aorta.

1. The effects of 12-O-tetradecanoyl 4 beta-phorbol 13-acetate (beta-TPA) on the inositol 1,4,5-trisphosphate (IP3) production, Ca2+ release from the intracellular Ca2+ stores and sensitization of contractile apparatus, induced by prostaglandin F2 alpha (PGF2 alpha) and U46619, a thromboxane A2-mimetic, were studied, using fura-2-loaded and -unloaded rat thoracic aortic strips. 2. Both eicosanoids had characteristic patterns of responses in Ca(2+)-free, 2 mM EGTA-containing solution (Ca(2+)-free solution). They induced transient increases in intracellular Ca2+ concentration ([Ca2+]i) without corresponding transient contraction, but produced delayed, sustained contraction, where [Ca2+]i was returned to the basal level. 3. Treatment with beta-TPA for 60 min reduced the eicosanoids-induced IP3 production, suggesting that the treatment inhibits PIP2 breakdown. 4. The treatment also attenuated [Ca2+]i transient induced by the eicosanoids, but not by caffeine (an IP3-independent releaser of stored Ca2+), in fura-2-loaded preparations incubated in Ca(2+)-free solution. 5. In contrast in the presence of beta-TPA, the sustained contractions evoked by the eicosanoids in Ca(2+)-free solution were potentiated, suggesting that the sites of actions of beta-TPA and the eicosanoids may differ from each other. 6. PGF2 alpha and U46619 utilize different and parallel signal transduction pathways to release Ca2+ by IP3 produced by PIP2 breakdown (beta-TPA-sensitive), and to increase the sensitivity of contractile apparatus, in which protein kinase C may not be involved (beta-TPA-insensitive).

Effects of ageing on nicotine-induced contraction and substance P-like materials release in guinea-pig bronchus.

1. Effects of ageing on nicotine-induced contraction and release of substance P-like materials in the bronchial preparations from guinea-pigs of different ages were studied. 2. The pD2 value (potency) of nicotine decreased with age from 10 to 100 weeks. The pD2 value of substance P did not change with age suggesting that substance P receptor mechanisms do not alter with age. 3. The amount of substance P-like materials released by nicotine (10(-4) M) decreased with age from 10 to 100 weeks, supporting our previous findings that nicotine contracts the guinea-pig bronchus through the release of substance P-like materials. 4. These results suggest that the age-related decrease in the pD2 value (potency) of nicotine is due to the reduction in the amount of substance P-like materials released by nicotine.

Ca2(+)-dependence of endothelin-1 induced contraction of rat tail artery.

1. In rat tail artery, endothelin-1(ET-1) caused an increase in the cytosolic free Ca2+ level ([Ca2+]i) followed by a relatively sustained but not steady-state contraction in Ca2+ containing solution. 2. In the early phase of the contraction, the rate of increase in [Ca2+]i, was much faster than that in muscle tension. However, after the increases in [Ca2+]i and muscle tension reached at their peaks, there was a good correlation between the changes of the two parameters. 3. ET-1 could not induce an apparent contractile response in Ca2(+)-free medium, notwithstanding it evoked a [Ca2+]i transient in this medium. 4. The results indicate that ET-1 induce a contraction of rat tail artery which is almost fully dependent on the [Ca2+]i changes, and may inhibit the Ca2(+)-sensitivity of the contractile filaments in the early phase of the contraction.

Effect of ageing on nicotine-induced contraction of guinea-pig bronchial preparation.

1. Nicotine-induced contraction of bronchial preparations isolated from 3, 6, 10, 50 and 100 week old guinea-pigs were studied. 2. Contractile responses to nicotine were inhibited by hexamethonium and a substance P antagonist but not by atropine or tetrodotoxin, suggesting that the nicotine-contraction was mediated through a release of substance P-like material(s). 3. The pD2 value (potency) of nicotine did not change with age from 3 to 10 weeks but decreased thereafter to 100 weeks, while no age-related change was found in the pD2 value of substance P. The pA2 value of hexamethonium did not change with age, suggesting that the affinity of drugs to nicotine receptors does not change with age. 4. These results suggest that possibility that age-related change in the pD2 value of nicotine is due to a change in the amount of nicotine receptors but not to a change in the affinity of nicotine to its receptors.

Stimulation of rat liver growth by a 1,3-dithiole derivative, KZ-1026.

The oral administration of 2-(1,3-dithiol-2-ylidene)-1-phenyl-1,3-butanedione (KZ-1026) to normal rats at doses of 50, 100 and 200 mg/kg/day for 3 days accelerated liver enlargement in association with a dose-dependent increase in the total amounts of protein, RNA and DNA in the liver. The liver weight at 24 hr after the third dose of 200 mg/kg reached 174% of the control. With respect to the effect on liver enlargement, KZ-1026 differed from phenobarbital, since KZ-1026, unlike phenobarbital, increased hepatic DNA content without significant effects on P-450 and aminopyrine-N-demethylase. Incorporation of [3H]thymidine into liver DNA was stimulated by a single dose of KZ-1026 (200 mg/kg), and it peaked at 24 hr post dose (18 times the control), followed by an increase in the number of liver nuclei. Liver growth was also accompanied by an increasing hepatic reserve cell mass, expressed by the capacity of eliminating exogenous galactose from the blood stream. Pretreatment with KZ-1026 (200 mg/kg/day) for 3 days significantly improved the survival rate of subtotally hepatectomized rats from 39% to 78%. These findings indicate that KZ-1026 accelerates hepatocyte proliferation, resulting in an enhancement of liver functional mass in normal rats.