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Anti-Inflammatory and Antioxidative Effects of Camellia japonica on Human Corneal Epithelial Cells and Experimental Dry Eye: In Vivo and In Vitro Study

机译:山茶对人角膜上皮细胞和实验性干眼的抗炎和抗氧化作用:体内和体外研究

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Purpose: To analyze the anti-inflammatory and antioxidative effects of Camellia japonica (CJ) on human corneal epithelial (HCE) cells and its therapeutic effects in a mouse model of experimental dry eye (EDE). Methods: Camellia japonica extracts of varying concentrations (0.001%, 0.01%, and 0.1%) were used to treat HCE cells. Dichlorofluorescein diacetate (DCF-DA) and dihydroethidium (DHE) assays were performed. The production of peroxiredoxin (PRX) 1-6 and manganese-dependent superoxide dismutase (MnSOD) in HCE cells was assessed using Western blot analysis. Furthermore, eye drops containing 0.001%, 0.01%, or 0.1% CJ extract or a balanced salt solution (BSS) were applied to the EDE. Clinical parameters were measured 7 days after treatment. The levels of inflammatory markers and intracellular reactive oxygen species (ROS) were measured. Results: Treatment with 0.01% and 0.1% CJ extracts decreased apoptosis in HCE cells. In addition, band intensities of PRX 1, 4, and 5, as well as MnSOD, after hydrogen peroxide (H2O2) treatment showed a significant improvement after pretreatment with 0.01% and 0.1% CJ extracts. Mice treated with 0.1% CJ extract showed significantly improved clinical parameters when compared to those of the EDE control and BSS groups. A significant decrease in the levels of inflammatory markers and intracellular ROS was observed in the 0.01% and 0.1% CJ extract groups. Conclusions: Camellia japonica extracts promoted antioxidative protein expression and suppressed apoptosis in HCE cells. Furthermore, CJ extracts improved clinical signs of dry eye and reduced oxidative stress and the expression of inflammatory markers, suggesting that eye drops containing CJ extract could be used as an adjunctive treatment for dry eye.
机译:目的:分析山茶(CJ)对人角膜上皮(HCE)细胞的抗炎和抗氧化作用及其在实验性干眼(EDE)小鼠模型中的治疗作用。方法:使用不同浓度(0.001%,0.01%和0.1%)的山茶提取物处理HCE细胞。进行了二乙酸二氯荧光素(DCF-DA)和二氢乙啶(DHE)分析。使用蛋白质印迹分析评估HCE细胞中过氧化物酶(PRX)1-6和锰依赖性超氧化物歧化酶(MnSOD)的产生。此外,将含有0.001%,0.01%或0.1%CJ提取物或平衡盐溶液(BSS)的滴眼液应用于EDE。治疗后7天测量临床参数。测量了炎症标志物和细胞内活性氧(ROS)的水平。结果:用0.01%和0.1%CJ提取物处理可降低HCE细胞的凋亡。此外,过氧化氢(H2O2)处理后,PRX 1、4和5以及MnSOD的谱带强度在用0.01%和0.1%的CJ提取物进行预处理后显示出显着的改善。与EDE对照组和BSS组相比,用0.1%CJ提取物治疗的小鼠表现出明显改善的临床参数。在0.01%和0.1%的CJ提取物组中,炎症标志物和细胞内ROS的水平显着下降。结论:山茶提取物可促进HCE细胞抗氧化蛋白表达并抑制细胞凋亡。此外,CJ提取物改善了干眼症的临床体征,并减轻了氧化应激和炎症标志物的表达,表明含CJ提取物的滴眼液可用作干眼症的辅助治疗。

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