Effect of Hesperidin on Expression of Inducible Nitric Oxide Synthase in Cultured Rabbit Retinal Pigment Epithelial Cells

  • Luo Xiaoting
  • Zeng Xiangyun
  • Li Shumei
  • Dong Minghua
  • Xiong Liang
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 664)

Abstract

Objective: To study the effect of hesperidin on expression of inducible nitric oxide synthase (iNOS) in cultured rabbit retinal pigment epithelial (RPE) cells under the condition of high glucose in vitro.

Method: Hesperidin was extracted from Pericarpium Citri Reticulatae by ultrasound and ethanol precipitation and was detected qualitatively by high performance liquid chromatogram. The third to fifth primary cultured rabbit RPE were selected. The cells were divided into 6 groups including the control group cultured in DMEM, the model group cultured in DMEM containing 33 mmol/L glucose without any drug and four experimental groups which were exposed to hesperidin at the concentration of 10, 20, 40 and 80 mg/L at 37°C under 5% CO2 for 2 h and then cultured in DMEM containing 33 mmol/L glucose. The proliferation of RPE was measured by the MTT assay. The levels of NO produced were measured by spectrophotometry. The changes of iNOS expressed in RPE cells were determined with immunohistochemistry.

Results: The growth rate of RPE cells was associated with the concentration of hesperidin. NO production induced by high glucose was significantly inhibited by hesperidin. iNOS expression in hesperidin-treated group was decreased compared with the control group (p <0.001).

Conclusion: Hesperidin can increase the proliferation of rabbit RPE cells, and inhibit the level of NO and iNOS expression, so hesperidin can protect rabbit RPE cells.

Keywords

Estrogenic Respiration Retina Nitrite Trypsin 

References

  1. Civan MM, Marano CW, Matschinsky FW et al (1994) Prolonged incubation with elevated glucose inhibits the regulatory response to shrinkage of cultured human retinal pigment epithelial cells. J Membr Biol 139(1):1–13PubMedGoogle Scholar
  2. Hong J, Yuan Z, Shuai J et al (2007) High glucose induce production of iNOS by human retinal pigment epithelium cells through activation of the p38 signal pathway. Acta Universitatis Medicinalis Nanjing (Natural Science) 27(9):970–973Google Scholar
  3. Izumi N, Nagaoka T, Mori F et al (2006) Relation between plasma nitric oxide levels and diabetic retinopathy. Jpn J Ophthalmol 50(5):465–468CrossRefPubMedGoogle Scholar
  4. Kanaze FI, Gabrieli C, Kokkalou E et al (2003) Simultaneous reversed-phase high-performance liquid chromatographic method for the determination of diosmin, hesperidin and naringin in different citrus fruit juices and pharmaceutical formulations. J Pharm Biomed Anal 33(2):243–249CrossRefPubMedGoogle Scholar
  5. Leal EC, Manivannan A, Hosoya K et al (2007) Inducible nitric oxide synthase isoform is a key mediator of leukostasis and blood-retinal barrier breakdown in diabetic retinopathy. Invest Ophthalmol Vis Sci 48(11):5257–5265CrossRefPubMedGoogle Scholar
  6. Luo X, Li S (2008) Progress in Biologic Activity Effects of Hesperidin. J Gannan Med Univ 28(2):301–304Google Scholar
  7. Madsen-Bouterse SA, Kowluru RA (2008) Oxidative stress and diabetic retinopathy: pathophysiological mechanisms and treatment perspectives. Rev Endocr Metab Disord 9(4):315–327CrossRefPubMedGoogle Scholar
  8. Rao YK, Fang SH, Tzeng YM (2008) Antiinflammatory activities of flavonoids and a triterpene caffeate isolated from Bauhinia variegata. Phytother Res 22(7):957–962CrossRefPubMedGoogle Scholar
  9. Roufail E, Soulis T, Boel E et al (1998) Depletion of nitric oxide synthase-containing neurons in the diabetic retina: reversal by aminoguanidine. Diabetologia 41(12):1419–1425CrossRefPubMedGoogle Scholar
  10. Sakata K, Hirose Y, Qiao Z et al (2003) Inhibition of inducible isoforms of cyclooxygenase and nitric oxide synthase by flavonoid hesperidin in mouse macrophage cell line. Cancer Lett 199(2):139–145CrossRefPubMedGoogle Scholar
  11. Shashi W, Tapas CN (1999) Nitric oxide synthase immunoreactivity in the developing and adult human retina. J Biosci 24(4):483–490CrossRefGoogle Scholar
  12. Toda N, Nakanishi-Toda M (2007) Nitric oxide: ocular blood flow, glaucoma, and diabetic retinopathy. Prog Retin Eye Res 26(3):205–238CrossRefPubMedGoogle Scholar
  13. Tommasini S, Calabrò ML, Stancanelli R et al (2005) The inclusion complexes of hesperetin and its 7-rhamnoglucoside with (2-hydroxypropyl)-beta-cyclodextrin. J Pharm Biomed Anal 39(3–4):572–580CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Luo Xiaoting
    • 1
  • Zeng Xiangyun
    • 2
  • Li Shumei
    • 3
  • Dong Minghua
    • 3
  • Xiong Liang
    • 3
  1. 1.Department of Biochemistry and Molecular BiologyGannan Medical CollegeGanzhouChina
  2. 2.Department of Ophthalmology of the 1st Affiliated HospitalGannan Medical CollegeGanzhouChina
  3. 3.Department of Preventive MedicineGannan Medical CollegeGanzhouChina

Personalised recommendations