Experimental Sciences
Distribution of lanosterol synthase and lanosterol in cornea, lens and retina tissue of rats
Lihua Kang, Xinyue Shen, Mei Yang, Guowei Zhang, Junfang Zhang, Bai Qin, Ling Yang, Nan Hu, Huaijin Guan
Published 2017-03-10
Cite as Chin J Exp Ophthalmol, 2017, 35(3): 201-206. DOI: 10.3760/cma.j.issn.2095-0160.2017.03.003
Abstract
BackgroundResearches showed that triterpenoids, with a similar structure to lanosterol, has therapeutical effect on many systemic diseases, and lanosterol was determined to have a therapeutical effect on cataract recently.However, how the lanosterol plays effects on other eye diseases is still unelucidated.Understanding the distribution of lanosterol in ocular tissue is helpful for us to elucidate the relationship of lanosterol with eye diseases.
ObjectiveThis study attempted to investigate the distribution of lanosterol synthase (LSS) and lanosterol in cornea, lens and retina tissue of rats and offer a basis for the targeting treatment of eye diseases.
MethodsFifteen SPF male SD rats were sacrificed by excessive anesthesia to obtain the eyeballs.The relative expressions of LSS protein and gene in the cornea, lens and retina tissue of the rats were detected by Western blot and reverse transcription (RT)-PCR, respectively.Immunofluorescence staining technology was used to locate the distribution of LSS in cornea, lens and retina tissue.The contents of lanosterol in the cornea, lens and retina tissue were analyzed by liquid chromatograph mass spectrometer (LC-MS).
ResultsNo LSS protein and mRNA was expressed in the retinal tissue in normal rats.The mean relative expression of LSS protein in the lens and cornea was 0.43±0.05 and 0.25±0.03, respectively, showing a significant difference between them (t=-5.35, P<0.01). The relative expression of LSS mRNA was 0.51±0.04 and 0.29±0.02 in the lens and cornea, respectively, with a stronger expression in the lens in comparison with the cornea (t=-8.34, P<0.01). Immunofluorescence staining showed that LSS primarily located in corneal epithelial layer, stromal layer and endothelial layer as well as lens epithelial cells and shallow cortex layer and hardly expressed in retina, and no co-expression of LSS with the neuron marked by NeuN and the Müller cell marked by glutamine synthetase (GS) in retinal tissue.LC-MS analysis revealed that the contents of lanosterol in lens and cornea was (24.37±2.91)ng/mg and (5.31±0.58)ng/mg, respectively, with a significant difference between them (t=-11.13, P<0.01).
ConclusionsLSS and lanosterol extensively distribute in cornea and lens of normal rats, but not in retina tissue.These results offer new strategies for the target treatment of relevant eye diseases.
Key words:
Lanosterol/metabolism; Lanosterol synthase; Cornea; Lens, crystalline; Retina; Cataract; Rat, SD
Contributor Information
Lihua Kang
Eye Institute, Affiliated Hospital of Nantong University, Nantong 226001, China
Xinyue Shen
Mei Yang
Guowei Zhang
Junfang Zhang
Bai Qin
Ling Yang
Nan Hu
Huaijin Guan
