Wu Man-Ru, Lin Cheng-Hui, Ho Jau-Der, Hsiao George, Cheng Yu-Wen
School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei, Taiwan.
Department of Ophthalmology, Taipei Medical University, Taipei, Taiwan.
Cell Physiol Biochem. 2018;51(1):63-79. doi: 10.1159/000495162. Epub 2018 Nov 15.
BACKGROUND/AIMS: Blue light-emitting diode light (BLL)-induced phototoxicity plays an important role in ocular diseases and causes retinal degeneration and apoptosis in human retinal pigment epithelial (RPE) cells. Cistanche tubulosa extract (CTE) is a traditional Chinese medicine with many beneficial protective properties; however, few studies have examined the ocular protective roles of CTE. In this study, we investigated the mechanisms underlying the effects of CTE on BLL-induced apoptosis in vitro and in vivo.
RPE cells were applied in the current in vitro study and cell viability was determined by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis-related protein expression was determined by western blot analysis and immunofluorescence staining. Brown Norway rats were used to examine exposure to commercially available BLL in vivo. Hematoxylin and eosin staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and western blot assays were used to examine retinal morphological deformation.
CTE significantly inhibited hydrogen peroxide-, tert-butyl hydroperoxide-, sodium azide-, and BLL-induced RPE damage. Further, CTE reduced the expression of apoptotic markers such as cleaved caspase-3 and TUNEL staining after BLL exposure by inactivating apoptotic pathways, as shown via immunofluorescent staining. In addition, CTE inhibited the BLL-induced phosphorylation of c-Jun N-terminal kinase, extra signal-related kinases 1/2, and p38 in RPE cells. In vivo, the oral administration of CTE rescued 60-day periodic BLL exposure-induced decrements in retinal thickness and reduced the number of TUNEL-positive cells in the brown Norway rat model.
CTE is a potential prophylactic agent against BLL-induced phototoxicity.
背景/目的:蓝光发光二极管光(BLL)诱导的光毒性在眼部疾病中起重要作用,并导致人视网膜色素上皮(RPE)细胞发生视网膜变性和凋亡。管花肉苁蓉提取物(CTE)是一种具有多种有益保护特性的传统中药;然而,很少有研究探讨CTE的眼部保护作用。在本研究中,我们调查了CTE在体外和体内对BLL诱导的凋亡作用的潜在机制。
在当前的体外研究中使用RPE细胞,并通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法测定细胞活力。通过蛋白质印迹分析和免疫荧光染色测定凋亡相关蛋白的表达。使用棕色挪威大鼠在体内检查暴露于市售BLL的情况。苏木精和伊红染色、末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)和蛋白质印迹分析用于检查视网膜形态变形。
CTE显著抑制过氧化氢、叔丁基过氧化氢、叠氮化钠和BLL诱导的RPE损伤。此外,如免疫荧光染色所示,CTE通过使凋亡途径失活,降低了BLL暴露后凋亡标志物如裂解的半胱天冬酶-3的表达和TUNEL染色。此外,CTE抑制RPE细胞中BLL诱导的c-Jun氨基末端激酶、细胞外信号调节激酶1/2和p38的磷酸化。在体内,口服CTE挽救了60天周期性BLL暴露诱导的棕色挪威大鼠模型视网膜厚度的减少,并减少了TUNEL阳性细胞的数量。
CTE是一种潜在的预防BLL诱导的光毒性的药物。