ABSTRACT
Objective:To study the effect and mechanism of angiotensin type 1 receptor (AGTR1) blocker olmesartan (OMS) on the apoptosis of human Tenon capsule fibroblasts (HTF).Methods:Tenon capsule tissues were obtained from patients during strabismus surgery in the Second Affiliated Hospital of Xi'an Jiaotong University.Primary HTF were cultured by explant culture.Primary cells were identified by vimentin immunofluorescence staining and flow cytometry.The fibrosis model of HTF was established using 10 ng/ml transforming growth factor-β2 (TGF-β2). The cells were divided into normal control group cultured in culture medium, TGF-β2 group in culture medium containing TGF-β2, TGF-β2+ OMS group in culture medium containing TGF-β2 and OMS, and OMS group in culture medium containing OMS, and were cultured for 48 hours.Cell apoptosis was detected by flow cytometry with annexin V/PI staining.The early apoptosis, late apoptosis, and total apoptosis rates were analyzed.The protein expression of procaspase-9, cleaved caspase-9, bax and bcl-2 in the mitochondrial apoptosis pathway was detected by Western blot.The activity of lactate dehydrogenase (LDH) and superoxide dismutase (SOD) was detected by colorimetry.The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Xi'an Jiaotong University (No.2019-014).Results:Primary HTF were successfully isolated and cultured.The cultured cells were long spindle-shaped and positive for vimentin.The expression rate of vimentin in the primary cells was greater than 99%.A statistically statistical difference was found in the early apoptosis rate, late apoptosis rate, and total apoptosis rate among the four groups ( F=24.92, 3.96, 41.82; all at P<0.05). The early and total apoptosis rates were significantly higher in TGF-β2+ OMS group than normal control group and TGF-β2 group, and the late apoptosis rate in TGF-β2+ OMS group was significantly higher than that of normal control group (all at P<0.05). There were statistically significant differences in cleaved caspase-9/procaspase-9, bax, and bax/bcl-2 among the four groups ( F=4.40, 7.98, 4.61; all at P<0.05). The bax/bcl-2 expression was significantly increased in TGF-β2+ OMS group in comparison with normal control group, and the expressions of cleaved caspase-9/procaspase-9, bax, and bax/bcl-2 were significantly elevated in TGF-β2+ OMS group compared with TGF-β2 group (all at P<0.05). LDH activity in the normal control group, TGF-β2 group, TGF-β2+ OMS group and OMS group was (783.99±79.97), (913.16±196.86), (2 529.06±240.21), and (2 134.29±138.96) μmol/(min·L), respectively, showing a statistically significant difference ( F=24.95, P<0.05). Compared with normal control group and TGF-β2 group, LDH activity in TGF-β2+ OMS group was increased, and the differences were statistically significant (both at P<0.05). SOD activity in the normal control group, TGF-β2 group, TGF-β2+ OMS group and OMS group was (50.35±0.97), (41.61±4.56), (28.88±3.26), and (37.61±4.83) μmol/(min·L), respectively, showing a statistically significant difference ( F=5.71, P<0.05). SOD activity was reduced in TGF-β2+ OMS group compared with normal control group and TGF-β2 group, reduced in OMS group compared with normal control group, and the differences were statistically significant (all at P<0.05). Conclusions:AGTR1 blocker OMS can promote the apoptosis of HTF effectively.Mitochondrial apoptosis pathway mediated by bax/bcl-2/caspase-9 and oxidative stress pathway are the potential mechanisms that OMS regulates the apoptosis of HTF.
ABSTRACT
Objective:To establish and preliminarily validate a nomogram model for predicting the risk of retinal vein occlusion (RVO).Methods:A retrospective clinical study. A total of 162 patients with RVO (RVO group) diagnosed by ophthalmology examination in The Second Affiliated Hospital of Xi'an Jiaotong University from January 2017 to April 2022 and 162 patients with age-related cataract (nRVO group) were selected as the modeling set. A total of 45 patients with branch RVO, 45 patients with central RVO and 45 patients with age-related cataract admitted to Xi'an Fourth Hospital from January 2022 to February 2023 were used as the validation set. There was no significant difference in gender composition ratio ( χ2=2.433) and age ( Z=1.006) between RVO group and nRVO group ( P=0.120, 0.320). Age, gender, blood routine (white blood cell count, hemoglobin concentration, platelet count, neutrophil count, monocyte count, lymphocyte count, erythrocyte volume, mean platelet volume, platelet volume distribution width), and four items of thrombin (prothrombin time, activated partial thrombin time, fibrinogen, and thrombin time) were collected in detail ), uric acid, blood lipids (total cholesterol, triglyceride, high-density lipoprotein, low-density lipoprotein, lipoprotein a), hypertension, diabetes mellitus, coronary heart disease, and cerebral infarction. Neutrophil/lymphocyte ratio and platelet/lymphocyte ratio were calculated. The single logistic regression was used to analyze the clinical parameters of the two groups of patients in the modeling set, and the stepwise regression method was used to screen the variables, and the column graph for predicting the risk of RVO was constructed. The Bootstrap method was used to repeated sample 1 000 times for internal and external verification. The H-L goodness-of-fit test and receiver operating characteristic (ROC) curve were used to evaluate the calibration and discrimination of the nomogram model. Results:After univariate logistic regression and stepwise regression analysis, high density lipoprotein, neutrophil count and hypertension were included in the final prediction model to construct the nomogram. The χ2 values of the H-L goodness-of-fit test of the modeling set and the validation set were 0.711 and 4.230, respectively, and the P values were 0.701 and 0.121, respectively, indicating that the nomogram model had good prediction accuracy. The area under the ROC curve of the nomogram model for predicting the occurrence of post-stroke depression in the modeling set and the verification set was 0.741 [95% confidence interval ( CI) 0.688-0.795] and 0.741 (95% CI 0.646-0.836), suggesting that the nomogram model had a good discrimination. Conclusions:Low high density lipoprotein level, high neutrophil count and hypertension are independent risk factors for RVO. The nomogram model established based on the above risk factors can effectively assess and quantify the risk of post-stroke depression in patients with cerebral infarction.