Evaluation of pharmacological activities and assessment of intraocular penetration of an ayurvedic polyherbal eye drop (Itone™) in experimental models

Abstract

BackgroundThe 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.Materials and methodsAntiangiogenic 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.ResultsPHF 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.ConclusionThe significant antiangiogenic and anti-inflammatory activities evinced by the PHF merits further investigation for ocular neovascular and inflammatory diseases in humans.

Antiangiogenic, wound healing and antioxidant activity ofCladosporium cladosporioides(Endophytic Fungus) isolated from seaweed (Sargassum wightii)

“…The different level of activities of the two fungi isolated from the same species may be attributed to the different chemical composition of each extract. This is in agreement with the observations of Gul et al (2013). …”

Molecular Mechanisms of Casticin Action: an Update on its Antitumor Functions

“…As shown in Figure 1, accumulated data indicate that casticin was isolated from many plant species such as, namely Vitex agnus castus (Choudhary et al, 2009;Mesaik et al, 2009;Webster et al, 2011;Righeschi et al, 2012;Hogner et al, 2013), Daphne genkwa (Xie et al, 2011), Achillea millefolium (Haidara et al, 2006;Csupor-Loffler et al, 2009), Ficus microcarpa , Vitex rotundifolia (Ono et al, 2002;Hu et al, 2007b;Shen et al, 2009;Ye et al, 2010;Koh et al, 2011), Fructus viticis (Hu et al, 2007b;Guan et al, 2010;Chen et al, 2011b;Yang et al, 2011;Zeng et al, 2012;Zhou et al, 2013a), Vitex negundo (Diaz et al, 2003;Kunwar et al, 2010;Velpandian et al, 2013), Crataegus pinnatifida (Kao et al, 2005), Pavetta crassipes (Mali and Dhake, 2011), Nelsonia canescens, Butea frondosa Koen , Dalbergia odorifera (Mali and Dhake, 2011), Bryonia laciniosa (Aggarwal et al, 2011), Citrus unshu (Mali and Dhake, 2011;Nagoor et al, 2011), Centipeda minima (Mali and Dhake, 2011), Clausena excavate (Manosroi et al, 2005), Croton betulaster (de Sampaio e Spohr et al, 2010;Freitas et al, 2011), Dimorphandra mollis (Freitas et al, 2011), Artemisia abrotanum L. (Hernandez et al, 1999), Artemisia annua L …”

“…It inhibited eosinophil migration and activity of chemokines and adhesion of molecules involved in the inflammatory process of asthma by suppressing the NF-κB pathway (Koh et al, 2011). Casticin's biological effects have been reported in wide spectrum of indications (Table 1), including inflammation (Lin et al, 2007b;Koh et al, 2011;Lee et al, 2012), asthma (Koh et al, 2011), tumor (Ono et al, 2002;Hu et al, 2007b;Shen et al, 2009;Ye et al, 2010;Koh et al, 2011), pre-menstrual syndrome (Hu et al, 2007b;Webster et al, 2011), immunomodulation (You et al, 1998), headache (Choudhary et al, 2009;Mesaik et al, 2009;Webster et al, 2011;Righeschi et al, 2012;Hogner et al, 2013), rheumatoid arthritis (You et al, 1998), conjunctivitis (Remberg et al, 2004), trachoma, gonorrhoea, and toothache (Diaz et al, 2003;Kunwar et al, 2010;Velpandian et al, 2013).…”

“…Casticin (3', 5-dihydroxy-3, 4', 6, 7-tetramethoxyflavone) is one of the bioactive flavonoids obtained from polyphenol plants, which are composed of a wide variety of molecules that are classified into several categories, according to their chemical type, such as phenolic acids, flavonoids, stilbenes, and lignans (Siasos et al, 2013 ). Casticin is a main active compound in roots, aerial parts, seed, wood, stems, leaves and fruits of variety of plants ( Figure 1 and Table 1), has been reported to be responsible for a wide spectrum of biological and pharmacological activities including immunomodulatory (Mesaik et al, 2009;Ling et al, 2012), anti-hyperprolactinemia (Hu et al, 2007b;Ye et al, 2010), anti-tumor (Haidara et al, 2006;Shen et al, 2009;Ling et al, 2012;Zeng et al, 2012), neuroprotetective (Ling et al, 2012), anti-inflammatory (Lin et al, 2007b;Choudhary et al, 2009;Velpandian et al, 2013) and analgesic activities (Lee et al, 2012).…”

Role of Apo E and superoxide dismutase in patients with obstructive lung diseases

“…In fact, when this homeostasis is disrupted and there are more reactive species than antioxidants several diseases can develop. 15 Recent studies indicate that an increased oxidative stress is related to the pathogenesis of COPD, 1,[16][17][18] and contribute this to the fact that COPD patients produce more H 2 O 2 compared to "healthy". 19 Another study hypothesized that loss of asthmatic SOD activity is due to greater susceptibility to oxidative inactivation.…”