ABSTRACT
AIM: To investigate the expression and clinical significance of interferon regulatory factor 4(IRF4)and soluble suppression of tumorigenesis 2(sST2)in conjunctival epithelial cells and tears of patients with dry eye.METHODS: A total of 94 patients with dry eye who admitted to our hospital from January 2019 to December 2021 were selected as the dry eye group, and 97 physical examiners who underwent ophthalmic examination were selected as the control group at the same time. The conjunctival epithelial cells and tears of the subjects were collected, and the clinical indicators, including tear film break-up time(BUT), corneal fluorescein staining(CFS)score, and Schirmer Ⅰ test(SⅠt)were recorded. The levels of IRF4 and sST2 in conjunctival epithelial cells were detected by quantitative real-time polymerase chain reaction(qRT-PCR), and the levels of IRF4 and sST2 in tears were detected by enzyme-linked immunosorbent assay(ELISA). Pearson method was used to analyze the correlation between IRF4 and sST2 levels in conjunctival epithelial cells and tears and clinical indicators of dry eye patients.RESULTS: The levels of IRF4 and sST2 in conjunctival epithelial cells and tears in dry eye group before treatment were significantly higher than those in control group(P<0.001). The levels of IRF4 and sST2 in conjunctival epithelial cells and tears of dry eye patients at 4wk after treatment were significantly lower than those before treatment(P<0.001). The BUT and SⅠt of dry eye patients increased significantly at 4wk after treatment, and the CFS score decreased significantly(P<0.001). The levels of IRF4 and sST2 in conjunctival epithelial cells and tears of dry eye patients before treatment were positively correlated with CFS score before treatment and negatively correlated with BUT and SⅠt before treatment(P<0.001).CONCLUSION: The levels of IRF4 and sST2 in conjunctival epithelial cells and tears of patients with dry eye are increased, and are significantly correlated with BUT, SⅠt and CFS scores, which has potential to become a new therapeutic target for dry eye.
ABSTRACT
AIM: To explore the expression and significance of forkhead box class O3(FOXO3)and interleukin-2(IL-2)in conjunctival epithelial cells and tears of patients with dry eye(DE).METHODS:A perspective study. A total of 106 DE patients who accepted from March 2019 to March 2021 were prospectively gathered, and 85 healthy subjects in the same period were selected as the control group. The level of FOXO3 in the conjunctival epithelial cells and tear fluid was measured by real-time fluorescent quantitative PCR(qRT-PCR)method; The level of IL-2 in the sample was measured by enzyme-linked immunosorbent(ELISA)method; The changes in clinical indicators of the ocular surface such as break-up time(BUT), Schirmer Ⅰtest(SⅠt), cornea fluorescein staining(CFS)in DE patients before and after treatment were analyzed; The correlation between the levels of FOXO3 and IL-2 in the conjunctival epithelial cells and tears of DE patients and the relationship between the two and clinical indicators were analyzed by Pearson correlation analysis.RESULTS:Compared with the control group, the level of FOXO3 in conjunctival epithelial cells and tear fluid in the DE group was obviously reduced, and the level of IL-2 was obviously increased(all P<0.01). Compared with before treatment, the level of FOXO3 in conjunctival epithelial cells and tear fluid of DE patients was obviously up-regulated, and the level of IL-2 was obviously down-regulated(all P<0.05). Pearson correlation analysis showed that the levels of FOXO3 and IL-2 in conjunctival epithelial cells and tear fluid were obviously inversely correlated(r=-0.531, -0.469, all P<0.01). After treatment, BUT and SⅠt indexes of DE patients increased compared with before treatment, while CFS decreased(all P<0.01). The level of FOXO3 in conjunctival epithelial cells of DE patients was obviously directly correlated with BUT and SⅠt(r=0.431, 0.457, all P<0.01), and it was obviously inversely correlated with CFS(r=-0.469, P<0.01), and the level of IL-2 was obviously inversely correlated with BUT and SⅠt(r=-0.416, -0.447, all P<0.01), and it was obviously directly correlated with CFS(r=0.424, P<0.01); tear FOXO3 was positively correlated with BUT and SⅠt(r=0.421, 0.443, all P<0.01), and it was negatively correlated with CFS(r=-0.474, P<0.01), and IL-2 was negatively correlated with BUT and SⅠt(r=-0.408, -0.429, all P<0.01), and it was positively correlated with CFS(r=0.419, P<0.01).CONCLUSION: the level of FOXO3 in conjunctival epithelial cells and tears of DE patients is decreased, and the level of IL-2 is increased. The two of which are closely related to the ocular surface indicators of patients. They are expected to become laboratory auxiliary indicators for clinical monitoring and prognostic evaluation of DE.
ABSTRACT
PURPOSE: To investigate the toxicity of a short-term application of timolol maleate, dorzolamide, and benzalkonium chloride (BAC) on human conjunctival epithelial cells in vitro. METHODS: Chang's conjunctival epithelial cell line was treated for 5 min, 15 min, 30 min, and 60 min with various concentrations of timolol, dorzloamide, or BAC, and then examined 4 hrs or 24 hrs later. Cell viabilities were assessed by MTT assay. The expressions of various cytokines by timolol maleate, dorzolamide, and BAC treatment in human conjunctival epithelial cells were evaluated using ELISPOT. RESULTS: BAC significantly decreased survival of conjunctival epithelial cells in a dose and time dependent manner compared with timolol and dorzolamide. Inflammatory cytokines, IL-6 and IL-8, were highly expressed in conjunctival epithelial cells treated with timolol, dorzolamide, and BAC. CONCLUSIONS: The present study suggests that increased expression of inflammatory markers, IL-6 and IL-8 might explain the ocular surface disorder in patients receiving antiglaucoma medication.
Subject(s)
Humans , Benzalkonium Compounds , Cell Survival , Cytokines , Enzyme-Linked Immunospot Assay , Epithelial Cells , Interleukin-6 , Interleukin-8 , TimololABSTRACT
PURPOSE: Tanilast has been clinically used for various allergic diseases. Recently, it has also been found to inhibit excessive scarring in wound healing process. The purpose of this study is to investigate the inhibitory effect of Tranilast on the proliferation of human conjunctival epithelial cells and subconjunctival fibroblasts. METHODS: Human conjunctival epithelial cells and subconjunctival fibroblasts were exposed for 24 hours to Tranilast 0.05, 0.2, 0.4, 0.8, and 1.6 mg/ml. MTT based calorimetric assay was performed to assess the metabolic activity and inhibition of cellular proliferation. RESULTS: As the concentration of Tranilast increased, the absorbance of spectrometer decreased. Inhibitory effect of cellular proliferation was stronger in subconjunctival fibroblasts than in conjunctival epithelial cells. The inhibitory effect of celluar proliferation in conjunctival epithelial cells was 75%, 66%, 64%, 59%, and 39% at 0.05, 0.2, 0.4, 0.8, and 1.6 mg/ml, respectively (LD50 was 0.9 mg/ml). In subconjunctival fibroblasts, the inhibitory effect of celluar proliferation was 66%, 62%, 47%, 33%, and 23% at 0.05, 0.2, 0.4, 0.8, and 1.6 mg/ml, respectively (LD50 was 0.3 mg/ml). CONCLUSION: Tranilast may prevent excessive proliferation of both human conjunctival epithelial cells and subconjunctival fibroblasts. This inhibitory effect on cellular proliferation was higher in subconjunctival fibroblasts than in conjunctival epithelial cells, suggesting the possibility of being used after glaucoma filtering surgery or pterygium excision.