Protective effect of bilberry (Vaccinium myrtillus L.) extracts on cultured human corneal limbal epithelial cells (HCLEC)

The use of bilberry (Vaccinium myrtillus L.) as a food and medicine for improving human vision has a long history all over the world. However, there is lack of convincing evidence from rigorous clinical trials or scientific research. This study investigated the effects of different concentrations of bilberry extracts on the cell viability, cell cycle and the expression of hyaluronic acid and glycosaminoglycans of cultured human corneal limbal epithelial cells. The data showed that bilberry extracts had no cytotoxicity to the corneal limbal epithelial cells at a wide range of concentrations (10−9–10−4 M, equalized to the content of cyanidin‐3‐O‐glucoside). Bilberry extract (10−6, 10−5 and 10−4 M) increased cell viability after 48 h incubation. The number of cells decreased in G0/G1 phase and increased prominently in S and G2/M phases after treatment with bilberry extracts at a high concentration (10−4 M). The expression of glycosaminoglycans increased prominently after incubation with bilberry extracts (10−7 and 10−4 M) for 48 h while no significant changes were observed for the expression of hyaluronic acid. The results indicated that bilberry extract may be beneficial for the physiological renewal and homeostasis of corneal epithelial cells. Copyright © 2010 John Wiley & Sons, Ltd.

[1]  M. Sasaki,et al.  Enhancing effects of sericin on corneal wound healing in rat debrided corneal epithelium. , 2009, Biological & pharmaceutical bulletin.

[2]  M. Dutot,et al.  High molecular weight hyaluronan decreases UVB-induced apoptosis and inflammation in human epithelial corneal cells , 2009, Molecular vision.

[3]  L. Zanatta,et al.  Flavonoids: prospective drug candidates. , 2008, Mini reviews in medicinal chemistry.

[4]  T. Linsenmayer,et al.  Nuclear ferritin‐mediated protection of corneal epithelial cells from oxidative damage to DNA , 2008, Developmental dynamics : an official publication of the American Association of Anatomists.

[5]  F. Tost,et al.  Lubricating agents differ in their protection of cultured human epithelial cells against desiccation. , 2008, Medical science monitor : international medical journal of experimental and clinical research.

[6]  L. Morbidelli,et al.  Hydrocaffeic and p-coumaric acids, natural phenolic compounds, inhibit UV-B damage in WKD human conjunctival cells in vitro and rabbit eye in vivo , 2008, Free radical research.

[7]  Z. Wang,et al.  Potassium-Chloride Cotransporter Mediates Cell Cycle Progression and Proliferation of Human Corneal Epithelial Cells , 2007, Cell cycle.

[8]  H. Navsaria,et al.  Hyaluronic acid: the scientific and clinical evidence. , 2007, Journal of plastic, reconstructive & aesthetic surgery : JPRAS.

[9]  Brigitte Graf,et al.  Bilberry (Vaccinium myrtillus) anthocyanins modulate heme oxygenase-1 and glutathione S-transferase-pi expression in ARPE-19 cells. , 2007, Investigative ophthalmology & visual science.

[10]  J. Ge,et al.  Establishment of a corneal epithelial cell line spontaneously derived from human limbal cells. , 2007, Experimental eye research.

[11]  J. Ulrichová,et al.  Cytoprotective effect of a bilberry extract against oxidative damage of rat hepatocytes , 2007 .

[12]  Yu Yao,et al.  Protective activities of Vaccinium antioxidants with potential relevance to mitochondrial dysfunction and neurotoxicity. , 2007, Neurotoxicology.

[13]  A. Theocharis,et al.  Wild blueberry (Vaccinium angustifolium) consumption affects the composition and structure of glycosaminoglycans in Sprague-Dawley rat aorta. , 2006, The Journal of nutritional biochemistry.

[14]  R. Prior,et al.  Fate of anthocyanins and antioxidant capacity in contents of the gastrointestinal tract of weanling pigs following black raspberry consumption. , 2006, Journal of agricultural and food chemistry.

[15]  M. Rahman,et al.  Superoxide radical- and peroxynitrite-scavenging activity of anthocyanins; structure-activity relationship and their synergism , 2006, Free radical research.

[16]  N. Kolosova,et al.  [Dietary supplementation with bilberry extract prevents macular degeneration and cataracts in senesce-accelerated OXYS rats]. , 2008, Advances in gerontology = Uspekhi gerontologii.

[17]  P. Maher,et al.  Flavonoids protect retinal ganglion cells from oxidative stress-induced death. , 2005, Investigative ophthalmology & visual science.

[18]  C. Santos-Buelga,et al.  Antioxidant properties of prepared blueberry (Vaccinium myrtillus) extracts. , 2005, Journal of agricultural and food chemistry.

[19]  C. Joo,et al.  Wnt-7a Up-regulates Matrix Metalloproteinase-12 Expression and Promotes Cell Proliferation in Corneal Epithelial Cells during Wound Healing* , 2005, Journal of Biological Chemistry.

[20]  K. Nakanishi,et al.  Anthocyanins Protect Against A2E Photooxidation and Membrane Permeabilization in Retinal Pigment Epithelial Cells¶ , 2005, Photochemistry and photobiology.

[21]  M. Gnädinger,et al.  Synthesis of sulfated glycosaminoglycans by the three cell types of the rabbit cornea in culture , 1976, Experientia.

[22]  E. Ernst,et al.  Anthocyanosides of Vaccinium myrtillus (bilberry) for night vision--a systematic review of placebo-controlled trials. , 2004, Survey of ophthalmology.

[23]  C. Sen,et al.  Anti-angiogenic, Antioxidant, and Anti-carcinogenic Properties of a Novel Anthocyanin-Rich Berry Extract Formula , 2004, Biochemistry (Moscow).

[24]  E. Kowalczyk,et al.  Anthocyanins in medicine. , 2003, Polish journal of pharmacology.

[25]  S. Jockusch,et al.  A2E-epoxides Damage DNA in Retinal Pigment Epithelial Cells , 2003, The Journal of Biological Chemistry.

[26]  M. Hirayama,et al.  Stimulatory effect of cyanidin 3-glycosides on the regeneration of rhodopsin. , 2003, Journal of agricultural and food chemistry.

[27]  F. Guglielmi,et al.  Protection against ultraviolet B-induced oxidative DNA damage in rabbit corneal-derived cells (SIRC) by 4-coumaric acid. , 2003, Toxicology.

[28]  C. Sen,et al.  Anti-angiogenic Property of Edible Berries , 2002, Free radical research.

[29]  R. Prior,et al.  Anthocyanins are absorbed in glycated forms in elderly women: a pharmacokinetic study. , 2001, The American journal of clinical nutrition.

[30]  B. Shukitt-Hale,et al.  Polyphenolics enhance red blood cell resistance to oxidative stress: in vitro and in vivo. , 2000, Biochimica et biophysica acta.

[31]  S. Saika,et al.  Role of Lumican in the Corneal Epithelium during Wound Healing* , 2000, The Journal of Biological Chemistry.

[32]  I. Zagon,et al.  Cellular dynamics of corneal wound re-epithelialization in the rat III. Mitotic activity , 1999, Brain Research.

[33]  H. Durchschlag,et al.  Radiation effects on eye components , 1999 .

[34]  T. Chikama,et al.  Hyaluronan facilitates corneal epithelial wound healing in diabetic rats. , 1997, Experimental eye research.

[35]  M. Chapman,et al.  Antioxidant action of Vaccinium myrtillus extract on human low density lipoproteins in vitro: initial observations , 1997, Fundamental & clinical pharmacology.

[36]  T. Tsuru,et al.  [Quantitative analysis of glycosaminoglycans in tear fluids during corneal epithelial wound healing in rabbits]. , 1994, Nippon Ganka Gakkai zasshi.

[37]  U. Stenevi,et al.  Endogenous hyaluronan in corneal disease. , 1994, Investigative ophthalmology & visual science.

[38]  W. Knudson,et al.  Hyaluronan‐binding proteins in development, tissue homeostasis, and disease , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[39]  U. Stenevi,et al.  Evidence of hyaluronic acid and hyaluronic acid binding sites on human corneal endothelium , 1992, Journal of cataract and refractive surgery.

[40]  R. Beuerman,et al.  Flow cytometry measurements of the DNA content of corneal epithelial cells during wound healing. , 1991, Investigative ophthalmology & visual science.

[41]  T. Mosmann Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[42]  B. Gabetta,et al.  Qualitative and quantitative evaluation of vaccinium myrtillus anthocyanins by high-resolution gas chromatography and high-performance liquid chromatography , 1983 .

[43]  I. Constable,et al.  Vitreous structure. I. Distribution of hyaluronate and protein. , 1972, Investigative ophthalmology.