園芸学部研究業績紹介(目次)

<p>Multivariable analysis to determine prognostic factors for surgical failure of 2^nd operated trabeculectomy using Cox proportional hazards regression models among the successful 1^st operated eyes.</p

第1084回千葉医学会例会・第21回千葉精神科集談会

<p>The comparison of the successful 2^nd operated eyes versus failed 2^nd operated eyes among successful 1^st operated eyes, Criterion C: n = 30.</p

STRING protein interaction analysis.

STRING analysis of 11 genes, expression changes were confirmed by RT-qPCR, and their predicted functional partners. The color of each edge shows the type of relationship in the following manner: light blue = “from curated databases,” dark purple = “experimentally determined,” green = “text mining,” black = “co-expression,” and light purple = “protein homology”.</p

Flowchart for narrowing down the genes for which expression levels were altered upon 2ME2 addition.

Probes without gene names were removed (21,282 genes remaining), genes with signal values of <5 were excluded (6,640 genes remaining), genes exhibiting more than two-fold increase between the Control and Galactose groups as well as decrease between the Galactose and 2ME2 groups were included (80 genes remaining), and genes of unknown function, ribosomal-protein genes, and genes for which primer design was difficult were excluded (43 genes remaining). Of these, 11 genes exhibited more than 10% expression decrease between the Galactose and 2ME2 groups compared with amount of increase expression between the Control and Galactose groups by RT-qPCR, which were consistent with the microarray analysis.</p

RT-qPCR analysis of genes with altered expression levels.

RT-qPCR on 43 genes selected from the microarray analysis, Results are shown as target gene mRNA levels normalized by Gapdh mRNA levels. Data are expressed as the mean ± SE. Asterisks indicate more than a 10% decrease in expression between the galactose and 2ME2 groups compared with the amount of increase between the control and galactose groups.</p

List of 43 genes for which RT-qPCR was performed.

Cataract is an eye disease, in which the lens becomes opaque, causing vision loss and blindness. The detailed mechanism of cataract development has not been characterized, and effective drug therapies remain unavailable. Here, we investigated the effects of Hypoxia-inducible factor 1 (HIF-1) inhibitors using an ex vivo model, in which rat lenses were cultured in galactose-containing medium to induce opacity formation. We found that treatment with the HIF-1 inhibitors 2-Methoxyestradiol (2ME2), YC-1, and Bavachinin decreased lens opacity. Microarray analysis on 2ME2-treated samples, in which opacity was decreased, identified genes upregulated by galactose and downregulated by inhibitor treatment. Subsequent STRING analysis on genes that showed expression change by RT-qPCR identified two clusters. First cluster related to the cytoskeleton and epithelial-mesenchymal transition (EMT). Second cluster related to the oxidative stress, and apoptosis. ACTA2, a known marker for EMT, and TXNIP, a suppressor of cell proliferation and activator of apoptosis, were present in each cluster. Thus, suppression of EMT and apoptosis, as well as activation of cell proliferation, appear to underlie the decrease in lens opacity.</div

Results of protein-protein interaction analysis.

Results of STRING analysis of 11 genes whose expression variation was confirmed by RT-qPCR (https://version-11-5.string-db.org/). Organisms selected were Homo sapiens. The color of each edge shows the type of relationship in the following manner: light blue = "from curated databases"; dark purple = "experimentally determined"; green = "text mining"; black = "co-expression"; and light purple = "protein homology”. (PDF)</p

Effect of 2ME2 on lens opacity.

(A) Schematic representation of the SD rat lens experiment. (B) Rat lenses were cultured in medium containing 30 mM galactose for 2–3 days (upper panel). After image acquisition, DMSO as a vehicle control or 10 μM, 20 μM, or 40 μM 2ME2 in DMSO were added to the galactose-containing medium, and culture was continued for 2–3 days. The number of days indicated in each panel represents the total days of incubation, “q” indicates samples used for RT-qPCR, and “M” indicates samples used for microarray analysis. (C) The level of lens opacity with and without 2ME2, as well as the change in opacity before and after addition of the inhibitor, were calculated [17]. Data are expressed as the mean ± SE. The two additional samples used for the quantification are shown in S2 Fig.</p

Lens photographs used for opacity quantification.

Upper panel shows lens photographs before and lower panel shows lens photographs after addition of inhibitor. In addition to the lens photographs in Fig 1B, a total of three samples were used for opacity quantification. Days in the upper left panel indicates total incubation time, "q" indicates samples RT-qPCR, and "M" indicates samples used for microarray analysis. (PDF)</p

List of 186 genes that were increased more than 2-fold from control to galactose and decreased more than 1.5-fold from galactose to 2ME2.

The ten columns on the right of Gene name show the signal value (log2) in each sample. Average indicates the mean value of replicate samples. The column for Control vs Galactose indicates the number of fold increase in expression from Control to Galactose. The column of Galactose vs. 2ME2 shows the number of fold decrease in expression from Galactose to 2ME2. (XLSX)</p