Improvement effect of gemigliptin on salivary gland dysfunction in exogenous methylglyoxal-injected rats.

The symptom of hyposalivation associated with hypofunction of the salivary glands is a common feature of diabetes. Inadequate saliva production can cause tissue damage in the mouth, making it susceptible to infections and leading to oral health diseases. Previous studies have highlighted the harmful effects of methylglyoxal (MGO) and MGO-derived advanced glycation end products (AGEs) in diabetes. In this study, we investigated the protective effects of gemigliptin, a dipeptidyl peptidase-4 (DPP-4) inhibitor, against MGO-induced salivary gland dysfunction. MGO treatment of immortalized human salivary gland acinar cells induced apoptosis via reactive oxygen species (ROS)-mediated pathways, but this effect was mitigated by gemigliptin. In vivo experiments involved the simultaneous administration of MGO (17.25 mg/kg) with aminoguanidine (100 mg/kg) and gemigliptin (10 and 100 mg/kg) daily to rats for two weeks. Gemigliptin increased the saliva volume and amylase levels in MGO-injected rats. Gemigliptin reduced the DPP-4 activity in both the salivary glands and serum of MGO-injected rats. Furthermore, gemigliptin exerted anti-glycation effects by reducing the accumulation of AGEs in the saliva, salivary glands, and serum and suppressing the expression of the receptor for AGEs. These actions protected the salivary gland cells from ROS-mediated apoptosis. Overall, gemigliptin protected the salivary gland cells from ROS-mediated cell death, reduced the accumulation of amylase and mucins in the salivary glands, and enhanced the salivary function by upregulating aquaporin 5 expression, and it exerted protective effects against MGO-induced salivary gland dysfunction by enhancing the anti-glycation, antioxidant, and salivary secretion activities. Our findings suggest gemigliptin as a potential therapeutic for patients with salivary gland dysfunction caused by the complications of diabetes.

Effect of YC-1102 on the Improvement of Obesity in High-Fat Diet-Induced Obese Mice.

Obesity is one of the major risk factors for metabolic diseases worldwide. This study examined the effects of YC-1102, an extract derived from the roots of Rosa multiflora, on 3T3-L1 preadipocytes and high-fat diet (HFD)-induced obese mice. In vivo experiments involved the oral administration of YC-1102 (100, 150, and 200 mg/kg body weight) daily to mice for eight weeks. YC-1102 was found to downregulate the expressions of PPARγ and C/EBPα during adipogenesis, inhibiting adipocyte differentiation and upregulating the expression of PGC-1α for energy metabolism to enhance mitochondrial biogenesis and fatty acid oxidation. It has been shown that daily administration of YC-1102 to mice receiving a HFD prevented an increase in body weight and the accumulation of body fat. YC-1102 administration also reduced TG, TC, and LDL cholesterol levels, as well as glucose and leptin levels, and increased adiponectin levels, thus effectively inhibiting the metabolism of lipids. YC-1102-treated mice showed significant reductions in the mRNA expression of PPARγ and C/EBPα. The levels of PGC-1α involved in energy metabolism increased significantly in the YC-1102-treated mice when compared to the HFD-treated mice. According to the findings of this study, YC-1102 has a dual mechanism that reduces transcription factors that promote the differentiation of adipocytes and increases transcription factors that promote energy consumption.

The Effects of Sargassum horneri Extract and Fucoidan on Tear Hyposecretion and Ocular Surface Injury in Rats with Dry Eye Diseases.

Hyperosmotic stress caused by tear hyposection is a leading cause of dry eye disease. We investigated the prevention of dry eye disease in corneal epithelial cells and in rats that were induced to develop dry eye disease via unilateral excision of their exorbital lacrimal gland using Sargassum horneri extract (AB_SH) and its bioactive component fucoidan. Oral administration of AB_SH (250 mg/kg and 500 mg/kg) and fucoidan (100 mg/kg) was conducted for 7 days. In order to measure tear secretion, phenol red thread tear tests were performed along with corneal irregularity measurements. The apoptotic injury in the cornea and the lacrimal gland was evaluated using TUNEL staining. AB_SH and fucoidan were shown to suppress apoptosis and the expression of apoptosis-related proteins in human corneal epithelial cells under hyperosmotic conditions. Oral administration of AB_SH and fucoidan attenuated tear hyposecretion and corneal irregularity in the lacrimal gland-excised rats. In addition, AB_SH and fucoidan also reduced apoptosis in the cornea and lacrimal gland. This study suggests that S. horneri extract and fucoidan can effectively ameliorate dry eye disease by suppressing the apoptosis of ocular tissues.

A comparison of peripapillary vessel density between subjects with normal-tension glaucoma and primary open-angle glaucoma with similar extents of glaucomatous damage.

The purpose of this study was to identify differences in retinal microvasculature impairments between patients with normal-tension glaucoma (NTG) and those with primary open-angle glaucoma (POAG) with similar extents of structural and visual field damage. Participants with glaucoma-suspect (GS), NTG, POAG, and normal controls were consecutively enrolled. Peripapillary vessel density (VD) and perfusion density (PD) were compared among the groups. Linear regression analyses were performed to identify the relationship between VD, PD and visual field parameters. The VDs of the full areas were 18.3 ± 0.7, 17.3 ± 1.7, 16.5 ± 1.7, and 15.8 ± 2.3 mm-1 in the control, GS, NTG, and POAG groups, respectively (P < 0.001). The VDs of the outer and inner areas and the PDs of all areas also differed significantly among the groups (all P < 0.001). In the NTG group, the VDs of the full, outer, and inner areas were significantly associated with all visual field parameters including the mean deviation (MD), pattern standard deviation (PSD), and visual field index (VFI). In the POAG group, the VDs of the full and inner areas were significantly associated with PSD and VFI but not with MD. In conclusion, with similar degrees of retinal nerve fiber layer thinning and visual field damage in both groups, the POAG group showed a lower peripapillary VD and PD than the NTG group. VD and PD were significantly associated with visual field loss.

CHARACTERISTICS OF THE MACULAR MICROVASCULATURE IN IDIOPATHIC EPIRETINAL MEMBRANE PATIENTS WITH AN ECTOPIC INNER FOVEAL LAYER.

PURPOSE: To identify the characteristics of the retinal microvasculature in epiretinal membrane patients with ectopic inner foveal layer (EIFL). METHODS: Patients were classified into two groups: those without EIFL (Group 1) and those with EIFL (Group 2). The vessel density (VD), perfusion density (PD), and the foveal avascular zone (FAZ) parameters were compared using optical coherence tomography angiography. Linear regression analysis was performed to identify the optical coherence tomography angiography parameters associated with best-corrected visual acuity. RESULTS: The VD of the central area in Group 1 and Group 2 was 11.6 ± 3.3 and 17.2 ± 2.8 mm -1 , respectively ( P < 0.001), the PD of the central area was 21.7 ± 6.2 and 32.0 ± 5.5%, respectively ( P < 0.001), and the FAZ area was 0.24 ± 0.11 and 0.09 ± 0.08 mm 2 , respectively ( P < 0.001). Based on the linear regression analysis, the VD of the central area (B = 0.018, P = 0.003), the PD of the central area (B = 0.009, P = 0.004), and FAZ area (B = -0.489, P = 0.013) were significantly associated with best-corrected visual acuity in patients with epiretinal membrane. CONCLUSION: The VD and PD of the foveal area were significantly higher in patients with EIFL, and the FAZ area was lower in patients with EIFL than in those without EIFL. In addition, the VD and PD of the foveal area were negatively associated with best-corrected visual acuity, and the FAZ area was positively associated with best-corrected visual acuity in patients with epiretinal membrane.

Gemigliptin Improves Salivary Gland Dysfunction in D-Galactose-Injected Aging Rats.

Oral dryness is among the most common conditions experienced by the elderly. As saliva plays a crucial role in maintaining oral health and overall quality of life, the condition is increasingly taking its toll on a rapidly growing aging population. D-galactose (D-gal) stimulates their formation, which in turn cause oxidative stress and accelerate age-related decline in physical function. In this study, we observed a reduction in salivary secretion and amylase levels in aged rats injected with D-gal, confirming salivary gland dysfunction. Treatment with gemigliptin increased DPP-4 inhibition and GLP-1 levels in the salivary glands of aging rats and reduced the expression of AGEs and receptors for advanced glycation end products (RAGE). This effect was caused by the presence of additional reactive oxygen species (ROS) in the salivary glands of the examined rats. Gemigliptin's cytoprotective effect reduced amylase and mucin accumulation and increased AQP5 expression, which are important indicators of salivary gland function. In sum, gemigliptin was shown to improve D-gal-induced decline in the salivary gland function of aged rats through its anti-glycation and antioxidant activities. Gemigliptin shows promise as a treatment strategy for patients experiencing decreased salivary function associated with their advancing age.

Antioxidant Efficacy of Esculetin against Tert-Butyl Hydroperoxide-Induced Oxidative Stress in HEK293 Cells.

Esculetin is an antioxidant and anti-inflammatory compound derived from coumarin. Oxidative stress can cause overproduction of reactive oxygen species (ROS), which can lead to the development of chronic kidney failure. In this study, human embryonic kidney 293 (HEK293) cells were treated with tert-butyl hydroperoxide (t-BHP) to determine the antioxidant effects of esculetin. HEK293 cells were treated with t-BHP to validate changes in cell viability, ROS production, and apoptosis, and then treated with esculetin to evaluate the changes. Changes in mRNA and protein levels were analyzed using a proteome kit, PCR, and Western blotting. Esculetin improved HEK293 cell viability and reduced apoptosis caused by t-BHP-induced oxidative stress. At the mRNA and protein levels, esculetin decreased pro-apoptotic factor expression as well as increased anti-apoptotic factor expression. The antioxidant efficacy of esculetin was validated when it inhibited the apoptosis caused by t-BHP-induced oxidative stress in HEK293 cells.

Effect of Esculetin on Tert-Butyl Hydroperoxide-Induced Oxidative Injury in Retinal Pigment Epithelial Cells In Vitro.

Esculetin is a coumarin-derived compound with antioxidant and anti-inflammatory properties. The current study aims to evaluate the therapeutic implications of esculetin on retinal dysfunction and uncover the underlying mechanisms. Tert-butyl hydroperoxide (t-BHP) at a concentration of 300 µM was used to induce oxidative stress in human retinal pigment epithelial cell line (ARPE-19) cells. Esculetin at concentrations below 250 µM did not cause cytotoxicity to ARPE-19 cells. Cell viability analysis confirmed that t-BHP induced oxidative injury of ARPE-19 cells. However, ARPE-19 cells were protected from t-BHP-induced oxidative injury by esculetin in a concentration-dependent manner. As a result of the TUNEL assay to confirm apoptosis, esculetin treatment reduced the number of TUNEL-positive cells. Esculetin down-regulated the expression levels of Bax, Caspase-3, and PARP and up-regulated the expression level of Bcl2. Collectively, this study demonstrates that esculetin exerts potent antioxidant properties in ARPE-19 cells, inhibiting t-BHP-induced apoptosis under the regulation of apoptotic factors.

Esculetin has therapeutic potential via the proapoptotic signaling pathway in A253 human submandibular salivary gland tumor cells.

Esculetin is a natural lactone that is commonly derived from coumarins. According to previous experiments using human cancer cells, esculetin has potent antitumor activity; it also inhibits proliferation and induces the apoptosis of cancer cells. In the present study, the anti-proliferative effect of esculetin on the submandibular salivary gland tumor cell line, A253, was evaluated via in vitro and in vivo analyses. Furthermore, the anti-cancer effects of esculetin in A253 cells and a xenograft model of salivary gland tumors were determined using 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide and TUNEL assay, apoptosis protein array, quantitative polymerase chain reaction and western blot analysis. Esculetin (50-150 µM) was demonstrated to have an anti-proliferative effect in the A253 cell line in vitro; this observed effect was dependent on the dose and duration of treatment. Esculetin also increased the levels of Bax, cleaved caspase-3, cleaved-9 and cleaved poly (ADP-ribose) polymerase apoptosis-related proteins, and decreased the expression levels of the Bcl-2 anti-apoptotic protein. With respect to apoptosis regulation, esculetin significantly decreased the proliferation of tumor cells in a xenograft model (100 mg/kg/day) for 18 days. Overall, esculetin could be a potential oral anticancer drug against salivary gland cancer.

The Ganglion Cell-Inner Plexiform Layer Thickness/Vessel Density of Superficial Vascular Plexus Ratio According to the Progression of Diabetic Retinopathy.

Purpose: The purpose of this study was to identify the relationship between the gangion cell-inner plexiform layer (GC-IPL) and retinal vasculature in the context of the progression of diabetic retinopathy (DR). Methods: The subjects were divided into four groups according to DR stage: normal controls (group 1), patients with diabetes mellitus (DM) without DR (group 2), patients with mild or moderate nonprogressive DR (NPDR; group 3), and patients with severe NPDR (group 4). GC-IPL thickness, vessel density of superficial vascular plexus (SVD), and the GC-IPL/SVD ratio were compared among the groups. Results: A total of 556 eyes were enrolled; 288 in group 1, 140 in group 2, 76 in group 3, and 52 in group 4. The mean GC-IPL thicknesses were 83.57 ± 7.35, 82.74 ± 7.22, 81.33 ± 6.74, and 79.89 ± 9.16 µm in each group, respectively (P = 0.006). The mean SVDs were 20.40 ± 1.26, 19.70 ± 1.56, 18.86 ± 2.04, and 17.82 ± 2.04 mm-1 in each group, respectively (P < 0.001). The GC-IPL/SVD ratios were 4.11 ± 0.38, 4.22 ± 0.40, 4.36 ± 0.54, and 4.54 ± 0.55 in each group, respectively (P < 0.001). In Pearson's correlation analysis, DR stage was significantly correlated with the GC-IPL/SVD ratio (coefficient = 0.301; P < 0.001). Conclusions: As the DR stage progressed, the GC-IPL thickness tended to decrease, with the macular SVD showing a significant reduction. Additionally, the impairment of retinal vasculature was more prominent than GC-IPL thinning as DR progressed, which suggests that retinal vasculature changes may precede diabetic retinal neurodegeneration.

Effect of Aucubin-Containing Eye Drops on Tear Hyposecretion and Lacrimal Gland Damage Induced by Urban Particulate Matter in Rats.

Exposure to particulate matter is a causative factor of dry eye disease. We aimed to investigate the beneficial effect of eye drops containing aucubin on dry eye disease induced by urban particulate matter (UPM). Dry eye was induced in male SD rats (6 weeks old) by topical exposure to UPM thrice a day for 5 d. Eye drops containing 0.1% aucubin or 0.5% aucubin were topically administered directly into the eye after UPM exposure for an additional 5 d. Tear secretion was evaluated using a phenol red thread tear test and corneal irregularity. The oxidative damage in the lacrimal gland was evaluated using TUNEL and immunohistochemical staining. The topical administration of aucubin significantly attenuated UPM-induced tear hyposecretion (control group: 9.25 ± 0.62 mm, UPM group: 4.55 ± 0.25 mm, 0.1% aucubin: 7.12 ± 0.58 mm, and 0.5% aucubin: 7.88 ± 0.75 mm) and corneal irregularity (control group: 0.00 ± 0.00, UPM group: 3.40 ± 0.29, 0.1% aucubin: 1.80 ± 0.27, and 0.5% aucubin: 1.15 ± 0.27). In addition, aucubin also reduced the UPM-induced apoptotic injury of lacrimal gland cells induced by oxidative stress through the increased expression of HMGB1 and RAGE. These findings indicate that the topical administration of aucubin eye drops showed a beneficial effect against UPM-induced abnormal ocular changes, such as tear hyposecretion and lacrimal gland damage. Therefore, our results reveal the pharmacological activities of aucubin in dry eye disease.

Comparison of Each Retinal Layer Thicknesses between Eyes with Central Retinal Vein Occlusion and Normal Contralateral Eyes.

PURPOSE: To evaluate the difference in each retinal layer thickness in central retinal vein occlusion (CRVO) with resolved macular edema after intravitreal antivascular endothelial growth factor injection and normal contralateral eyes.. METHODS: Patients with ischemic and nonischemic CRVO whose macular edema resolved after intravitreal antivascular endothelial growth factor injections and did not recur for at least 6 months, and a normal contralateral eye were enrolled. Each retinal layer thickness between CRVO and normal contralateral eyes was compared according to Early Treatment Diabetic Retinopathy Study subfields using spectral domain optical coherence tomography. RESULTS: The thicknesses of outer nuclear layer, photoreceptor layer, and retinal pigment epithelium in central ring, ganglion cell layer, inner plexiform layer, outer nuclear layer, and photoreceptor layer in the inner ring, and ganglion cell layer in the outer ring of CRVO eyes were significantly thinner than those of normal contralateral eyes (all p < 0.05). Whereas, inner nuclear layer and outer plexiform layer thicknesses in central ring of CRVO eyes were 23.86 ± 8.8 and 25.76 ± 7.6 µm, respectively, which was significantly thicker than those of normal contralateral eyes (19.52 ± 7.7 and 22.76 ± 6.5 µm; p = 0.019 and p = 0.043, respectively). Additionally, the mean best-corrected visual acuity of CRVO eyes were significantly correlated with photoreceptor layer thickness in central ring (p = 0.005). CONCLUSIONS: In CRVO eyes with resolved macular edema, the outer retinal layers were thinner as well as inner retinal layers, whereas inner plexiform layer and outer nuclear layer were thicker than normal fellow eyes. Additionally, photoreceptor layer thickness in foveal area had a significant impact on visual acuity in CRVO.

The Weiss Ring, a Major Confounding Factor For Measurements of Peripapillary Retinal Nerve Fiber Layer Thickness: Short title: The Impact of the Weiss Ring On RNFL Thickness Measurements.

PURPOSE: To identify how the Weiss ring affects the measurement of mean and sectoral peripapillary retinal nerve fiber layer (pRNFL) thicknesses. DESIGN: Retrospective, cross-sectional study. METHODS: Subjects were divided into two groups: controls (control group) and subjects in which a Weiss ring was visible on optical coherence tomography fundus images (WR group). Mean and sectoral pRNFL thicknesses were compared between the two groups. RESULTS: A total of 205 eyes were enrolled: 131 eyes in the control group and 74 eyes in the WR group. The mean pRNFL thicknesses of the control group and WR group were 97.2 ± 6.7 µm and 94.6 ± 10.8 µm, respectively (P = .042). In sectoral thickness, the inferior sector of the WR group was 112.1 ± 23.2 µm, which was significantly thinner than that of the control group (125.5 ± 13.3 µm; P < .001). The Weiss ring was located in 10 eyes (13.5%) in the superior sector, 7 eyes (9.5%) in the temporal sector, 40 eyes (54.1%) in the inferior sector, and 17 eyes (23.0%) in the nasal sector. In analyses of reproducibility, the coefficient of variation and intraclass coefficient of the inferior sector measurement were 10.90% and 0.409, respectively, indicating low reliability of the measurement. CONCLUSIONS: Eyes with a Weiss ring showed thinner mean and inferior pRNFL thicknesses than normal controls, which would be a measurement error caused by the Weiss ring. This could be a major confounding factor for analyses of pRNFL changes, especially in glaucoma patients.

Short-term effect of anti-VEGF for chronic central serous chorioretinopathy according to the presence of choroidal neovascularization using optical coherence tomography angiography.

PURPOSE: To analyze the short-term therapeutic efficacy of intravitreal injection of bevacizumab (IVB) for chronic central serous chorioretinopathy (CSC) according to the presence of choroidal neovascularization (CNV) using optical coherence tomography angiography (OCTA). METHODS: A retrospective chart review was perfomed on cases of CSC with CNV (Group 1: n = 31) and an age-matched cases of CSC without CNV (Group 2: n = 30). The response to IVB was evaluated by changes in best-corrected visual acuity (BCVA), central macular thickness (CMT), choroidal thickness (CT), and pachyvessel diameter. Univariate and multivariate linear regression analyses were performed to identify factors associated with the visual outcome of chronic CSC with CNV after IVB. RESULTS: At baseline, the CT values differed significantly between Groups 1 and 2 (371.55 ± 67.09 vs. 417.33 ± 71.32 µm, p = 0.01). In Group 1, BCVA improved significantly (p < 0.001), and CMT (p < 0.001), CT (p = 0.001) and pachyvessel diameter (p = 0.045) decreased significantly, after IVB. In Group 2, only pachyvessel diameter (p = 0.001) was significantly smaller after IVB. Univariate analysis showed that the initial CT (B = 0.002, p = 0.026) and pachyvessel diameter (B = 0.002, p = 0.001) significantly affected visual outcome. In multivariate analysis, the initial pachyvessel diameter exhibited significant results (B = 0.002, p = 0.001). CONCLUSIONS: IVB showed less effective short-term outcomes in chronic CSC patients without CNV than in patients with CNV. In chronic CSC with CNV, the short-term visual outcome after IVB was better in patients with a thinner choroid and smaller pachyvessels.

High-level production of recombinant trypsin in transgenic rice cell culture through utilization of an alternative carbon source and recycling system.

Productivity of recombinant bovine trypsin using a rice amylase 3D promoter has been studied in transgenic rice suspension culture. Alternative carbon sources were added to rice cell suspension cultures in order to improve the production of recombinant bovine trypsin. It was demonstrated that addition of alternative carbon sources such as succinic acid, fumaric acid and malic acid in the culture medium could increase the productivity of recombinant bovine trypsin 3.8-4.3-fold compared to those in the control medium without carbon sources. The highest accumulated trypsin reached 68.2 mg/L on day 5 in the culture medium with 40 mM fumaric acid. The feasibility of repeated use of the cells for recombinant trypsin production was tested in transgenic rice cell suspension culture with the culture medium containing the combination of variable sucrose concentration and 40 mM fumaric acid. Among the used combinations, the combination of 1% sucrose and 40 mM fumaric acid resulted in a yield of up to 53 mg/L five days after incubation. It also increased 31% (W/W) of dry cell weight and improved 43% of cell viability compared to that in control medium without sucrose. Based on these data, recycling of the trypsin production process with repeated 1% sucrose and 40 mM fumaric acid supplying-harvesting cycles was developed in flask scale culture. Recombinant bovine trypsin could be stably produced with a yield of up to 53-39 mg/L per cycle during five recycling cycles.

Production of functional recombinant bovine trypsin in transgenic rice cell suspension cultures.

A synthetic bovine trypsinogen (sbTrypsinogen) was synthesized on the basis of rice-optimized codon usage via an overlap PCR strategy, prior to being expressed under the control of the sucrose starvation-inducible rice α-amylase 3D (RAmy3D) promoter. Secretion of trypsin into the culture medium was achieved by using the existing signal peptide. The plant expression vector was introduced into rice calli (Oryza sativa L. cv. Dongjin), mediated by Agrobacterium tumefaciens. The integration of the sbTrypsinogen gene into the chromosome of the transgenic rice callus was verified via genomic DNA PCR amplification, and sbTrypsin expression in transgenic rice suspension cells was confirmed via Northern blot analysis. Western blot analysis detected glycosylated proteins in the culture medium, having masses from 24 to 26 kDa, following induction by sugar starvation. Proteolytic activity of the rice-derived trypsin was confirmed by gelatin zymogram, and was similar to that of the commercial bovine-produced trypsin. The yields of sbTrypsin that accumulated in the transgenic rice cell suspension medium were 15 mg/L at 5 days after sugar starvation.