Biophysics, Poster

INVESTIGATIONS OF HUMAN EYE LENS BY IMMUNOHYSTOLOGICAL AND IMMUNOCYTOCHEMICAL METHODS IN PATIENTS WITH SENILE AND DIABETIC CATARACT

T.G.Kamenskih, A.S.Tishkova, V.A.Galanzha, Clinic of Eye Diseases and Department of Ophthalmology of Saratov State Medical University n.a. V.I.Razumovsky
A.B.Bucharskaya, G.N.Maslyakova, Saratov State Medical University n.a. V.I.Razumovsky
A.A.Shirokov, A.M.Burov, Institute of Biochemistry and Physiology Plants and Microorganisms of Russian Akademy of Sciense
A.N.Bashkatov, Saratov State University
E.A.Genina, Saratov State University

ABSTRACT

Abstract
Human lens pathology in different forms of the cataract was studied. 220 patients with a cataract (220 eyes) were included in this study. Morphological, immunohystological and immunocytochemical investigations of the lens capsule and lens substance (antibodies to α- (α-А and α-В), β-, γ- crystallines), spectroscopic analyses of the smears and morphometric analyses of the slices were performed.
More significant expression of alfa-B and beta-crystallines was found in the patients with diabetes mellitus comparatively with the patients with age-related cataract. Morphological differences were founded in the patients with cortical and nuclear forms of senile cataract and the diabetic cataract.
Material and Methods.
220 patients with a cataract (220 eyes): the main group is 120 patients with diabetes mellitus (120 eyes) including 11 patients with diabetic cataract (11 eyes), 109 patients with senile cataract (109 eyes) with diabetes mellitus; the control group is 100 patients (100 eyes) with senile cataract with no diabetes were examined
All patients underwent the morphological, immunohystological and immunocytochemical investigations of the lens capsule and lens substance (antibodies to α- (α-А and α-В), β-, γ- crystallines).Spectroscopic analyses of the smears of the lens substance using spectrophotometer Lambda-950 (Perkin Elmer USA) was held. Morphometric analyses of the lens slices with the help of an automatic colorimetric system for image analyses of histological slices Ariol SL50 (Genetix,UK).was performed..
The images were obtained using microscope Olympus BX 61 with digital camera U-CMAD3 with lenses PlanApo N 1.25х/0,04 and MPlanFL N 5х/0,15 for choosing working areas, as well as UPlanFL N 20х/0,5 for estimating immunopositive areas. Fence of the lens material and its capsule was made during microsurgical cataract operations : ultrasonic, microcoacsial phacoemulsification extracapsular and intracapsular cataract extraction, Intraocular lenses were implanted in patients during the operations.
Conclusion
During the spectroscopic and morphometric analyses immunohystological and immunocytochemical investigations of lens preparation of patients with diabetes mellitus with complicated diabetic cataract as well as senile cataract with diabetes mellitus more significant expression of alfa-B and beta-crystallines was found in the patients with diabetes mellitus comparatively with the patients with age-related cataract.
Morphological differences were founded in the patients with cortical and nuclear forms of senile cataract and the diabetic cataract. The necessity of investigation of the cataractogenes molecular mechanism with diabetes mellitus to search anticataract drugs was proved.
Results
Patients’ senile cataract thickness of the capsule with epithelium was 12-16 mkm: The capsule was lined with single layer flattened epithelium with large nuclei. The capsule had a homogenous appearance and a smooth outer layer.
Patients’ diabetic cataract capsule side was irregularly thickened. Its thickness was 16-20 mkm, the capsule epithelium was increased in size and its shape was approaching to cubic, had a polygonal form. In the epithelial cells one could see vacuolization. Schick-test performed more bright colour of lens capsule preparation of diabetes mellitus patients This indicates the increased accumulation of carbohydrates in it. Morphometric analyses of the slices using an automatic colorimetric system for image analyses of histological slices Ariol SL50 (Genetix,UK) was performed
The increase of positive β- crystallines (75,9%) and α-В crystallines (56,4%) expression areas was found with the diabetes mellitus patients comparatively with the patients with age-related cataract (47,1% и 37% accordingly). More significant expression of alfa-B and beta-crystallines was found in the patients with diabetes mellitus during immunohystological and immunocytochemical investigations. All patients showed negative or very weak γ- crystallines expression

Representing author

photo

Mrs. Antonina Tishkova

Saratov State Medical University n.a.V.I.Razumovsky
Russia

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