Corneal protective effects of novel tear substitutes containing sodium hyaluronate and dodecahydrosqualene, squalane, in a porcine dry eye model.

To develop a novel tear substitute (TS) containing sodium hyaluronate (SH) and dodecahydrosqualene (DHS, squalane), we improved the prescription of a previously developed TS containing saline, 0.5% SH and 1% castor oil (CO), which had corneal protective effects against 60-min desiccation in a porcine dry eye model and viscosity of 106.8 mPa·S. Fresh porcine eyes were treated with a TS containing saline, 0.1%, 0.25%, 0.3% or 0.5% SH, and 1% CO or 1%, 2.5% or 5% DHS, and TS-treated eyes were desiccated for up to 180 min. The corneal damage was evaluated by the staining score of methylene blue (MB), absorbance of MB extracted from the cornea, the staining density of lissamine green (LG) and histopathology. The viscosities of the examined TS were also measured. A saline/0.5% SH/1% DHS solution had corneal protective effects for 90 min under desiccation and a viscosity of 110.0 mPa·s. A TS with saline, 0.1%, 0.25% or 0.3% SH and 1% or 2.5% DHS did not have better protective effects than a saline/0.5% SH/1% DHS solution, although a saline/0.3% SH/5% DHS solution exhibited greater corneal protection against 180-min desiccation on MB and LG staining and histopathological examination, and its viscosity was 34.5 mPa·s, which was similar to the 29.5 mPa·s of 0.3% SH. The saline/0.3% SH/5% DHS solution is available as a novel 3-hr long-lasting TS containing mucinomimetic and liquid oil components to treat and relieve dry eye symptoms in animals and humans.

Involvements of PHI-nitric oxide and PACAP-BK channel in the sustained relaxation of mouse gastric fundus.

The roles of nitric oxide (NO) and K(+) channels in sustained relaxation induced by electrical field stimulation (EFS) in the presence of atropine and guanethidine were studied in circular muscle strips of mouse gastric fundus. In the wild-type mouse, N(G)-nitro-l-arginine (l-nitroarginine), a nitric oxide synthase inhibitor, significantly inhibited the sustained relaxation in addition to the rapid relaxation. The sustained relaxation in pituitary adenylate cyclase-activating peptide (PACAP)-knockout mouse, which was smaller than that of the wild-type mouse, was also inhibited by l-nitroarginine. l-Nitroarginine inhibited the relaxation induced by the peptide histidine isoleucine (PHI), but not that induced by PACAP. S-Nitroso-N-acetyl-dl-penicillamine (SNAP), a NO donor, -induced relaxation was not affected by PACAP(6-38). EFS-induced sustained relaxation was inhibited by iberiotoxin, a big conductance calcium-activated K(+) (BK) channel inhibitor, but not by apamin, a small conductance calcium-activated K(+) (SK) channel inhibitor, and glibenclamide, an ATP-sensitive K(+) channel inhibitor. The relaxation that remained after the iberiotoxin-treatment was significantly inhibited by l-nitroarginine. Iberiotoxin inhibited PACAP-induced relaxation, while it had no effect on both PHI- and SNAP-induced relaxation. Immunoreactivities to anti-BK channel and anti-PHI antibodies were found in the circular muscle and the myenteric plexus layers, respectively. These results suggest interplay between PHI and NO in the sustained relaxation of the mouse gastric fundus, and that BK channels are involved in the PACAP-component of the sustained relaxation.

PACAP provides colonic protection against dextran sodium sulfate induced colitis.

Pituitary adenylate cyclase-activating polypeptide (PACAP) plays a crucial role in immunity and inflammation. Our aim was to obtain insight in the role of PACAP in experimental colitis in mice and thus its possible role in inflammatory bowel disease. PACAP-deficient (PACAP-/-) mice and wild-type control mice were challenged by colitis-inducing agent, dextran sulfate sodium (DSS). We monitored clinical symptoms, intestinal morphology, and difference of cytokine production in the proximal and distal colon. After DSS administration, mortality was more severe in PACAP-/- mice versus wild-type control mice. The histological score and the disease activity index of PACAP-/- mice were significantly higher than those of wild-type control mice. In proximal colon, production of IL-1beta and IL-6 in PACAP-/- mice were significantly upregulated on day 8 after DSS administration, compared to wild-type control mice. In distal colon, furthermore, production of IFNgamma, IL-1beta, IL-6, IL-12, and KC were significantly higher in PACAP-/- mice than in wild-type control mice on day 4. Our findings indicate that PACAP regulates the production of pro-inflammatory cytokine in the experimental colitis.