leukotriene-b4 and 1-1-diphenyl-2-picrylhydrazyl

The polyherbal eye drop (Itone™) is a mixture of aqueous distillates of nineteen traditionally used ingredients that sum up to impart potency to the formulation and make it a useful adjunct in various ocular pathologies. However, as there have been no controlled experimental studies accounting to the above claim, therefore, the present study was designed to evaluate the polyherbal formulation (PHF) for antiangiogenic, anti-inflammatory, anticataract, antioxidant and cytotoxicity in addition to the evaluation of intraocular penetration of PHF in rabbit eyes using LC-MS/MS.. Antiangiogenic activity of the PHF was evaluated using in ovo chick chorio-allantoic membrane (CAM) assay and in vivo cautery induced corneal neovascularization assay in rats. Anticataract potential was evaluated using steroid induced cataract in developing chick embryos, sodium selenite induced cataract in rat pups and galactose induced cataract in rats. The antioxidant activity was evaluated using di-phenyl picryl hydrazyl (DPPH) radical scavenging assay. Anti-inflammatory activity was evaluated in vitro using inhibition of LTB4 formation in human WBCs and in vivo using carrageenan induced paw edema assay in rats. The cytotoxicity was evaluated against HeLa cancer cell lines using (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Furthermore evaluation of the intraocular penetration of the PHF was carried out in rabbit eyes via aqueous humor paracentesis and further analysis using LC-MS/MS.. PHF significantly inhibited VEGF induced proliferation of new blood vessels in CAM assay and inhibited the cautery induced corneal neovascularization in rats. Additionally, PHF showed noticeable delay in the progression of cataract in the selenite and galactose induced cataract models whereby the PHF treated lenses were graded for stages II and III respectively. However, the PHF did not show any anticataract activity in the hydrocortisone induced cataract model. Moreover, PHF exhibited anti-inflammatory activity whereby it showed 39.34% inhibition of LTB4 formation and significantly inhibited carrageenan induced paw edema in rats. Eight compounds of PHF viz. camphor, casticin, curcumin-II, quercetin, rosmarinic acid, γ-terpinene, β-pinene and dipentene exhibited transcorneal penetration in rabbit eyes.. The significant antiangiogenic and anti-inflammatory activities evinced by the PHF merits further investigation for ocular neovascular and inflammatory diseases in humans.

Topics: Angiogenesis Inhibitors; Animals; Anti-Inflammatory Agents; Aqueous Humor; Biphenyl Compounds; Blood Vessels; Carrageenan; Cataract; Chick Embryo; Cornea; Edema; Eye; Female; Galactose; HeLa Cells; Humans; Hydrocortisone; Inflammation; Lens, Crystalline; Leukocytes; Leukotriene B4; Male; Medicine, Ayurvedic; Models, Animal; Ophthalmic Solutions; Phytotherapy; Picrates; Plant Extracts; Rabbits; Rats; Rats, Wistar; Sodium Selenite; Steroids; Vascular Endothelial Growth Factor A

Erycristagallin, a pterocarpene isolated from Erythrina mildbraedii, was tested in vitro for its antioxidant properties on the stable 2,2-diphenyl-1-pycryl-hydrazyl (DPPH) free radical and on the arachidonic acid metabolism. In addition, erycristagallin was tested on different experimental models of inflammation, such as the acute and chronic inflammation induced by the application of 12-O-tetradecanoylphorbol 13-acetate (TPA) on mice and the phospholipase A(2)-induced mouse paw oedema test. In the carrageenan-induced mouse paw oedema test, the ethyl acetate extract obtained from E. mildbraedii showed anti-inflammatory activity, and erycristagallin was isolated as the active principle. In vivo, erycristagallin significantly inhibited the phospholipase A(2)-induced mouse paw oedema as well as the mouse ear oedema induced by TPA (ID(50)<10 microg/ear). Moreover, it significantly reduced the chronic inflammation and leukocyte infiltration induced by repeated application of TPA. In vitro, erycristagallin inhibited the arachidonic acid metabolism via the 5-lipoxygenase pathway in rat polymorphonuclear leukocytes (IC(50)=23.4 microM), but had no effect on cyclooxygenase-1 metabolism in human platelets, while showing antioxidant activity in the DPPH test. As with other phenolics, the anti-inflammatory activity of erycristagallin may be based on its capacity to inhibit the arachidonic acid metabolism via the 5-lipoxygenase pathway.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Arachidonic Acid; Biphenyl Compounds; Carrageenan; Ear; Edema; Erythrina; Female; Free Radical Scavengers; Heterocyclic Compounds, 4 or More Rings; Hindlimb; Isoflavones; Leukotriene B4; Mice; Phospholipases A; Picrates; Plant Extracts; Rats; Rats, Wistar

Of several lichen metabolites isolated from Parmelia nepalensis and Parmelia tinctorum, the didepsides atranorin (4) and diffractaic acid (5), as well as (+)-protolichesterinic acid (7), inhibited LTB4 biosynthesis in polymorphonuclear leukocytes. Ethyl hematommate (3) and (+)-usnic acid (1) were only weak inhibitors, while methyl beta-orcinolcarboxylate (2) and gyrophoric acid (6) were inactive at concentrations up to 60 microM. Redox properties of the compounds were evaluated in terms of inhibition of nonenzymatic lipid peroxidation in model membranes, reactivity against the stable free radical 2,2-diphenyl-1-picrylhydrazyl, and deoxyribose degradation as a measure of hydroxyl-radical generation. The results revealed that lichen metabolites neither acted as antioxidants against the peroxidation process in model membranes nor did they scavenge or produce free radicals, suggesting that the inhibitory effects on LTB4 biosynthesis was due to specific enzyme interaction rather than a nonspecific redox mechanism.

Topics: Animals; Bepridil; Biphenyl Compounds; Cattle; Deoxyribose; Free Radicals; In Vitro Techniques; Leukocytes; Leukotriene B4; Lichens; Lipid Peroxidation; Oxidation-Reduction; Phospholipids; Picrates

The effect of phenolic compounds in foodstuffs on histamine and leukotriene B4 (LTB4) release from rat peritoneal exudate cells and their antioxidative activity were examined to assess their antiallergenic activities. Among them, triphenols such as pyrogallol and gallic acid inhibited histamine release from the cells, but diphenols did not. On the other hand, o- and p-diphenols such as catechol and hydroquinone with strong antioxidative activity inhibited LTB4 release as strongly as pyrogallol, but an m-derivative resorcinol with weak antioxidative activity did not. Though carboxylated compounds and their noncarboxylated counterparts were antioxidative, the former exerted a much weaker inhibitory effect on the LTB4 release than the latter. In flavonols, only myricetin with a triphenolic B ring strongly inhibited histamine release, but all flavonols strongly suppressed LTB4 release irrespective of the number of OH groups in the B ring. Among flavonoids with an o-diphenolic B ring, flavonol and flavone with a C4-carbonyl group strongly inhibited LTB4 release, whereas the activity of anthocyan without C4-carbonyl was much weaker than the above compounds. These results suggest that triphenolic structure is essential for the inhibition of histamine release. On the other hand, antioxidative activity and membrane permeability of phenolic compounds seemed to be essential for the inhibition of LTB4 release. In addition, the C4-carbonyl group seemed to be important for strongly inhibiting LTB4 release.

Topics: Animals; Antioxidants; Bepridil; Biphenyl Compounds; Butylated Hydroxytoluene; Exudates and Transudates; Flavonoids; Free Radicals; Histamine Release; Leukotriene B4; Male; Peritoneum; Phenols; Picrates; Polymers; Rats; Rats, Wistar; Structure-Activity Relationship; Vitamin E