BRIEF DESCRIPTIONS OF DRAWINGS
FIG. 1 shows the effects of topical application of NOS inhibitors in hairless mice. Topical application of L-NAME (NOS inhibitor) or nNOS inhibitor accelerated the barrier recovery of hairless mice after tape stripping. One micromolar solution of each reagent was applied on one flank, using two points per flank and four animals per treatment. **p<0.001, ***p<0.0001 compared with control. The error bar shows SD.
FIG. 2 shows the barrier recovery in nNOS knock-out mice after barrier disruption. After tape stripping, the barrier recovery in nNOS−/− mice (n=4) was significantly faster than in wild-type mice (n=4). ***p<0.0001 compared with wild-type. The error bar shows SD.
FIG. 3 shows the effect of topical application of NO donor in hairless mice. Topical application of SNAP delayed the barrier recovery of hairless mice after tape stripping. One millimolar solution of SNAP was applied on one flank, using two points per flank and four animals per treatment. **p<0.001, ***p<0.0001 compared with control. The error bar shows SD.
FIG. 4 shows that NO was released from skin of hairless mice immediately after barrier disruption. The NO level was significantly increased by tape stripping. The increase was blocked by pre-incubation with nNOS inhibitor (100 μM), but not with iNOS inhibitor (100 μM). Number of animals: n=14 (control and tape) and n=12 (nNOS inhibitor and iNOS inhibitor). *p<0.05, **p<0.005, ***p<0.0005 compared with control. The error bar shows SD.
FIG. 5 shows the effects of topical application of inhibitors and activator of guanylyl cyclase (GC) in hairless mice. Topical application of a GC inhibitor and an NO-sensitive GC inhibitor accelerated the barrier recovery of hairless mice after tape stripping. One micromolar solution of each reagent was applied on one flank, using two points per flank and four animals per treatment. ***p<0.0001 compared with control. The error bar shows SD.
FIG. 6 shows that SNAP increased the intracellular calcium concentration in cultured keratinocytes. The increase was blocked by ODQ, while nifedipine (NIF) had no effect. The vertical axis shows the ratio of relative intensity (340 nm/380 nm) after treatment to that before treatment. The number of cells for each measurement was 50. ***p<0.0001 compared with control. The error bar shows SD.
FIG. 7 shows that SNAP increased the intracellular calcium concentration in the cultured keratinocytes in calcium-free medium. The vertical axis shows the ratio of relative intensity (340 nm/380 nm) after treatment to that before treatment. The number of cells for each measurement was 50. ***p<0.0001 compared with control. The error bar shows SD.
FIG. 8 shows the effect of topical application of nNOS inhibitor and ODQ on epidermal hyperplasia of hairless mice induced by barrier disruption under low environmental humidity. In (a), (b) and (c), representative sections are shown. Bars: 20 μm. Acetone treatment increased DNA synthesis in the epidermal basal layer (a: dark cells are BrdU-positive) and the increase was blocked by topical application of nNOS inhibitor (b) or ODQ (c). Bars: 20 μm. The levels of epidermal DNA synthesis and the epidermal thickness in the same experiment are shown in (d and e). The number of BrdU-positive cells and the epidermal thickness was increased by acetone treatment under dry conditions. The increase was blocked by the topical application of nNOS inhibitor and ODQ. ***p<0.0001 compared with control. The error bar shows SD.
Patent Application
20070232595
