研究目的
To determine the effects of lipoic acid (LA)-dependent disulfide reduction on mouse lens elasticity, to synthesize the choline ester of LA (LACE), and to characterize the effects of topical ocular doses of LACE on mouse lens elasticity.
研究成果
LA treatment led to a concentration-dependent decrease in lens protein disulfides concurrent with an increase in lens elasticity. Topical ocular treatment with LACE increased lens elasticity in mice, suggesting a potential treatment to restore accommodative amplitude in presbyopia.
研究不足
The study is limited to mouse models, and the applicability to human lenses and presbyopia treatment requires further investigation.
1：Experimental Design and Method Selection:
The study involved incubating mouse lenses in medium supplemented with selected levels of LA, measuring lens elasticity, and determining P-SH and PSSP levels in homogenates. LACE was synthesized and characterized by mass spectrometry and HPLC. Mice were treated with LACE, and lens elasticity was measured.
2：Sample Selection and Data Sources:
Eight-month-old mouse lenses (C57BL/6J) were used.
3：List of Experimental Equipment and Materials:
Includes coverslip method for elasticity measurements, HPLC system, mass spectrometry, and computer-controlled instrument for elasticity measurements.
4：Experimental Procedures and Operational Workflow:
Detailed steps include incubation of lenses, elasticity measurements, synthesis and characterization of LACE, and topical ocular treatment of mice.
5：Data Analysis Methods:
Data were analyzed using PRISM6 for Mac OS 10, including nonlinear regression analysis.
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Hank’s balanced salt solution
Invitrogen
Used for rinsing enucleated eyes.
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Retiga Exi camera
QImaging
Used for photographing lenses.
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Nikon TE200-U microscope
Nikon
Used for photographing lenses.
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Olympus E330 camera
Olympus
Used for photographing lenses.
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Adobe Photoshop
Adobe Systems, Inc.
Used for preparing composite contact prints of the photographs.
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PRISM6
GraphPad Software, Inc.
Used for statistical analysis.
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Agilent 1050 HPLC system
Agilent
Used for HPLC analysis.
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C-18 column
250 3 4.6 mm, 5 um
Alltech Associates
Used for sample separation in HPLC analysis.
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Coulochem II multielectrode electrochemical detector
ESA, Inc.
Used for detecting and measuring concentrations of redox active compounds.
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TSQ Quantum Discovery MAX triple stage mass spectrometer
Thermo Fisher Scientific
Used for tandem mass spectrometry studies.
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