WO2022145511A1 - Method for providing information for predicting diabetic cataract - Google Patents

Abstract

The present invention relates to a method for providing information for predicting diabetic cataract, the method measuring the concentration of 25-hydroxy vitamin D in aqueous humour obtained from an individual for the prediction. As higher concentration in comparison to that of a control group indicates a higher probability of diabetic cataract, the present invention can aid in diagnosing the disease.

Description

Information provision method for predicting diabetic cataract

The present invention relates to an information providing method for predicting diabetic cataract.

Vitamin D is a multifunctional molecule that plays important roles in a variety of biological functions, including the regulation of calcium homeostasis. For example, vitamin D has been shown to reduce inflammatory mediators and act as antioxidants. Numerous studies show an inverse relationship between vitamin D levels and chronic diseases associated with chronic inflammation, including diabetes, high blood pressure, heart disease, multiple sclerosis, schizophrenia, and rheumatoid arthritis.

Vitamin D is identified in two forms: D2 (ergocalciferol) and D3 (cholecalciferol). Vitamin D status is primarily determined by measuring serum concentrations of a total of 25-hydroxyvitamin D (25(OH)D), including two forms of vitamin D. In general, when the serum concentration is less than 20 ng/ml, it is judged to be vitamin D deficiency, and when the serum concentration is 20 to 30 ng/ml, it is judged to be vitamin D deficiency.

The aqueous humor is a clear liquid that occupies the anterior part of the eye, derived from plasma but differs in composition from serum, with lower protein and glucose concentrations and higher ascorbic acid concentrations than serum. In addition, the aqueous humor functions to supply oxygen and nutrients to the eye tissues and remove wastes. Since the lens is an avascular internal ocular organ that is supplied with oxygen and nutrients by aqueous humor, changes in the composition of the aqueous humor can lead to cataracts.

Previously, it was known that the serum 25-hydroxyvitamin D (25-hydroxyvitamin D, 25(OH)D) concentration was related to eye diseases, but the present invention measured the 25-hydroxyvitamin D concentration in the aqueous humor. The purpose of this study was to confirm the relationship with eye diseases.

1. A method of providing information for prediction of diabetic cataract comprising measuring the concentration of 25-hydroxyvitamin D in aqueous humor isolated from a subject with cataracts.

2. In the above 1, when the concentration is higher than that of the control, predicting that the likelihood of diabetic cataract is higher than that of the control; further comprising, an information providing method for predicting diabetic cataract.

3. The method of providing information for the prediction of diabetic cataract according to the above 2, wherein the control group is an senile cataract individual.

Through the present invention, it is possible to accurately diagnose a target patient by providing information on the predictability of diabetic cataract. Accordingly, it can further help to provide an accurate treatment method.

1 is a graphical representation of the correlation of 25(OH)D concentrations in serum and aqueous humor.

2 is a graph showing the concentration of 25(OH)D in aqueous humor in senile cataract patients and diabetic cataract patients.

Hereinafter, the present invention will be described in detail.

The present invention relates to an information providing method for prediction of diabetic cataract comprising the step of measuring the concentration of 25-hydroxyvitamin D (25-Hydroxyvitamin D, 25(OH)D) in aqueous humor isolated from a cataract individual. it's about

In the present invention, a cataract refers to a disease in which visual acuity is reduced due to a clouding of the sensory receptor, and includes, for example, diseases such as senile cataract and diabetic cataract.

The cataract individual refers to all individuals with cataract symptoms even if they are not diagnosed with cataracts or are not diagnosed with cataracts due to genetic or environmental factors, and may be, for example, mammals including humans, but is not limited thereto. .

In the present invention, the aqueous humor refers to a transparent liquid in the form of water that fills the space between the cornea and iris of the eye and between the iris and the lens, and is also called an aqueous humor or aqueous humor.

In the present invention, 25-hydroxyvitamin D (25-Hydroxyvitamin D, 25(OH)D) is a metabolite of vitamin D, 25-hydroxyvitamin D2 or vitamin D3 (cholesterol), which is a metabolite of vitamin D2 (ergocalciferol) 25-hydroxyvitamin D3, which is a metabolite of calciferol). One of these concentrations may be measured, each concentration may be measured, or the total concentration thereof may be measured.

In the present invention, the step of measuring the concentration of 25(OH)D can be generally used as long as it is a method for measuring the concentration of a vitamin or its metabolite, for example, by fluorescence emission or a concentration measurement kit can be used. However, the present invention is not limited thereto.

The information providing method of the present invention may further include a step of measuring the 25(OH)D concentration of the aqueous humor isolated from the individual and predicting that the likelihood of diabetic cataract is higher than that of the control group if the concentration is higher than that of the control group. have.

The present inventors confirmed that the 25(OH)D concentration of the aqueous humor in diabetic cataract individuals was higher on average compared to the senile cataract individuals, so that the 25(OH)D concentration in the aqueous humor and diabetic cataracts was positively correlated. confirmed that there is. Therefore, by measuring the 25(OH)D concentration of aqueous humor isolated from the individual, if the concentration is higher than that of the control, it can be predicted that the likelihood of diabetic cataract is higher than that of the control.

The comparison of the concentration may be to compare the concentration of 25-hydroxyvitamin D2, vitamin 25-hydroxyvitamin D3, or a total concentration thereof, but is not limited thereto.

The control group may be any one cataract individual, any one diabetic cataract or senile cataract individual, may be an average value of concentration measurements of one or more cataract individuals, and may be a concentration measurement of one or more senile cataract individual It may be an average value of the values, and may be an average value of concentration measurement values of one or more diabetic cataract individuals, but is not limited thereto.

Hereinafter, examples will be given to describe the present invention in detail.

Experimental subjects and methods

1. Experimental design

This study is a prospective study conducted at Gyeongsang National University Hospital in Jinju. From January to November 2017, fluid and serum samples were collected from 136 patients who underwent cataract surgery. This study was conducted with the approval of Gyeongsang National University IRB (2017-01-011), and all patients obtained written consent prior to participation.

2. Identification of patient groups and experimental conditions

After collecting the patient's medical history, all patients underwent ophthalmologic examinations, including best-corrected visual acuity and intraocular pressure measurements, slit lamp biomicroscopy, and fundus examination. The selected patients were divided into senile cataract and diabetic cataract patients. Patients with combined cataracts, including those with ocular trauma, uveitis, and cataracts due to long-term steroid use, and those who had previously undergone ophthalmic surgery were excluded. The diabetic cataract group included patients with well-controlled diabetes mellitus (hemoglobin A1C<7%) with nonproliferative diabetic retinopathy. Patients with proliferative diabetic retinopathy and those who underwent vitrectomy or intravitreal injection were excluded from the group, and patients with diabetes and other ocular diseases (eg, glaucoma or retinal disease) were excluded from the senile cataract group.

Cataract type and severity were graded using the Lens Opacity Classification System III. Anterior volume (ACV), axial anterior depth (ACD), and central corneal thickness (CCT) were measured using a Pentacam system (Oculus Inc., Wetzlar, Germany), and ocular vitality was measured with IOL Master.

The system (IOL Master 500, Carl Zeiss Meditec, Jena, Germany) measured the axial length (AL) using signals from the tear film and retinal pigment epithelium. Corneal endothelial cell number and morphology were investigated by CellChek XL (Konan Medical, Irvine, CA, USA), and retinal macular thickness was measured using spectralis OCT volume scan (Heidelberg Engineering, Heidelberg, Germany). Patients underwent a slit lamp test for tear film break-up time (TBUT) and Schirmer's test, and questionnaires were performed to assess occupation, average daily activity level, and vitamin D supplementation use.

All cataract surgeries were performed by experienced surgeons. After performing local anesthesia, before completing the cataract surgery before the main incision (2.2 m), about 150-200 μL of aqueous humor was collected according to the depth of the anterior chamber with a 30-gauze tuberculin syringe through the transparent cornea near the peritoneum.

experimental analysis

1. Experimental analysis

All aqueous humor samples were taken to the laboratory immediately after collection to measure vitamin D concentration, and serum samples were stored at -70°C until analysis. A total concentration of 25 (OH) D was measured using Cobas e602 and Elecsys Vitamin D Total Kits. The module (Roche Diagnostics, Mannheim, Germany) is an electrochemiluminescence assay that measures two forms of 25(OH)D (D2 and D3) in three steps. 25(OH)D bound in the first step is released from Vitamin D Binding Protein (VDBP), and 25(OH)D released in the second step binds to additional ruthenium labeled VDBP to form a complex to form In the third step, unbound ruthenium-labeled VDBP is removed by adding streptavidin-coated microparticles, and 25(OH)D is labeled with biotin. Applying a voltage to the electrode induces the ruthenium labeled VDBP-25(OH)D complex to emit chemiluminescence, which is measured with a photomultiplier tube. The signal generated was inversely proportional to the 25(OH)D concentration of the original sample. The measured concentration was standardized with the instrument-generated calibration curve, and the volume of each aqueous humor collected from the anterior chamber was about 150-200uL, and the actual volume analyzed with the Elecsys kit was 40uL.

2. Statistical analysis

Since most continuous data did not meet the assumptions of normality and equality of variance of the Shapiro-Wilk test and Leven's F test, respectively, categorical data are presented as numbers (%) and continuous data as medians (interquartile range, IQR). . Spearman's rank correlation analysis was used to evaluate the correlation between serum vitamin D concentration and aqueous humor. The aqueous humor and serum vitamin D concentrations were compared in the geriatric and diabetic cataract groups using the Mann-Whitney U test or the Kruskal-Wallis test.

Proportions of variables in the geriatric and diabetic cataract groups were compared using Fisher's exact test. To calculate the adjusted odds ratio (OR) of diabetic cataract to senile cataract, multivariate logistic regression analysis was performed. The adjustment variable selected for multivariate analysis is a variable with a P value of less than 0.15 and no multicollinearity in univariate analysis. In addition, clinically significant variables were included in the multivariate analysis for adjustment. The adjusted OR of aqueous humor vitamin D was calculated separately according to quartiles (Model 1) and continuous variables (Model 2).

Factors affecting the aqueous humor and serum vitamin D concentrations were evaluated by multivariate linear regression analysis using covariates with p values less than or equal to 0.15 in univariate analysis and those related to serum vitamin D concentrations. Since the concentrations of aqueous humor and serum vitamin D did not satisfy the normality assumption, the data were transformed to a partial rank probability method. All statistical analyzes were performed with SAS version 9.4 software (SAS Institute Inc., Cary, NC, USA), and bilateral p values <0.05 were considered statistically significant.

Experiment result

1. Characteristics of the test subject

A total of 136 patients were included in this study, including 87 geriatric cataract patients and 49 diabetic cataract patients. Their demographic and clinical characteristics are shown in Table 1.

[Table 1]

Vitamin D concentrations in aqueous humor (11.5 vs. 7.5 ng/mL, P = 0.010) and serum (20.1 vs. 14.2 ng/mL, P = 0.02) were higher in males than females (p=0.02). Also, the vitamin D concentration in the aqueous humor was higher in diabetic cataract patients than in geriatric cataract patients (12.6 vs. 8.8 ng/mL, P = 0.006). Tables 2 and 3 summarize the differences in each factor by cataract type. Most clinical and ophthalmic characteristics, such as gender, age, average daily outdoor time, occupation, and use of vitamin D supplementation, were associated with geriatric and diabetic cataract There were no significant differences between the patient groups (Table 2).

[Table 2]

Also, the prevalence of hypertension was higher in diabetic cataract patients than in geriatric cataract patients (p=0.012), and the proportion of axial length (AL) >24 mm patients was higher in the elderly cataract group than in the diabetic cataract group (p=0.012). 0.003, Table 3).

[Table 3]

2. Comparison of total 25(OH) concentration in aqueous humor and serum

The mean value (IQR) of the 25(OH)D concentration was 10.3 ng/mL in aqueous humor and 15.6 ng/mL in serum (Table 1). Measurements using the Cobas e602 module were possible when the sample volume was greater than 400uL.

There was no significant correlation between aqueous humor and serum 25(OH)D concentrations (p=0.381; FIG. 1). Aqueous 25(OH)D concentration was related to gender and cataract type, and serum 25(OH)D concentration was related to gender (Table 1). Mean concentration values were 4.0 ng/mL lower in women than in men (p=0.010), and 3.8 ng/mL lower in elderly patients than in diabetic cataract patients (p=0.006). The median values (IQR) of the aqueous humor vitamin D concentrations according to stages 0-1, 2-3, and 4-5 of nuclear color (NC) were 12.8, 10.4, and 8.2, respectively. In addition, the median values (IQR) of the aqueous humor vitamin D concentrations according to nuclear opalescence (NO) 0-1, 2-3, and 4-5 stages were 12.9, 10.4, and 8.4, respectively.

3. Comparison of ophthalmic test results according to types of cataracts and 25(OH) concentration in aqueous humor and serum

The ophthalmic test results and total 25(OH) concentrations in aqueous humor and serum were compared in diabetic cataract and senile cataract patient groups (Table 3). The ophthalmic test results were similar between the two groups, but the aqueous 25(OH)D concentration in the aqueous humor and serum showed that the aqueous 25(OH)D concentration was higher in diabetic cataracts (FIG. 2).

4. Multivariate Logistic Regression Analysis of Factors Associated with Diabetic Cataract

Factors included in multivariate logistic regression analysis were hypertension, AL, and 25(OH)D concentration in aqueous humor, all of which were significant in univariate analysis. As a result of analyzing the 25(OH)D concentration in the aqueous humor by quartiles, diabetic cataracts were significantly more frequent in Q4 than in Q1 subjects (Model 1: adjusted OR = 4.36, 95% confidence interval [CI] = 1.33-14.34, P = 0.015). Diabetic cataracts occurred less frequently in subjects with AL greater than 24 mm than in subjects with AL less than 24 mm (Model 1: adjusted OR = 0.24, 95% CI = 0.08-0.70, P = 0.009). In model 2, where 25(OH)D concentration was analyzed as a continuous variable, an increase in 25(OH)D concentration in the aqueous humor of 5 ng/ml increased OR of diabetic cataract patients by 1.2 (95% Cl=0.38-1.91). ; Table 4). There were no other factors associated with diabetic cataract.

[Table 4]

5. Results of Multivariate Linear Regression Analysis to Determine Factors Affecting Aqueous Aqueous and Serum 25(OH)D Concentrations

Factors affecting the 25(OH)D concentration in aqueous humor were gender and type of cataract. The 25(OH)D concentration in the aqueous humor was 2.79 ng/ml higher in men than in women and 2.68 ng/ml higher in diabetic cataract patients than in senile cataract patients (Table 5).

[Table 5]

In addition, patients who took vitamin D supplements had 7.8 ng/ml higher serum vitamin D concentrations than those who did not (Table 6).

[Table 6]

Claims (3)

1. Measuring the concentration of 25-hydroxyvitamin D (25-Hydroxyvitamin D, 25(OH)D) in aqueous humor isolated from a subject with cataracts; an information providing method for prediction of diabetic cataract comprising a.
2. The method according to claim 1, further comprising the step of predicting that the likelihood of diabetic cataract is higher than that of the control group when the concentration is higher than that of the control group.
3. The method according to claim 2, wherein the control group is an senile cataract individual.

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