Mei Xi-Yu, Zhou Ling-Yu, Zhang Tian-Yu, Lu Bin, Ji Li-Li
The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai Key Laboratory of Complex Prescription, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Int J Ophthalmol. 2017 Jun 18;10(6):870-877. doi: 10.18240/ijo.2017.06.07. eCollection 2017.
To observe the attenuation of ethanol extract of (SE) against diabetic retinopathy (DR) and its engaged mechanism.
C57BL/6J mice were intraperitoneally injected with streptozotocin (STZ, 55 mg/kg) for 5 consecutive days to induce diabetes. The diabetic mice were orally given with SE (100, 200 mg/kg) for 1mo at 1mo after STZ injection. Blood-retinal barrier (BRB) breakdown was detected by using Evans blue permeation assay. Real-time polymerase chain reaction (RT-PCR), Western blot and immunofluorescence staining were used to detect mRNA and protein expression. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum contents of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β.
SE (100, 200 mg/kg) reversed the breakdown of BRB in STZ-induced diabetic mice. The decreased expression of retinal claudin-1 and claudin-19, which are both tight junction (TJ) proteins, was reversed by SE. SE decreased the increased serum contents and retinal mRNA expression of TNF-α and IL-1β. SE also decreased the increased retinal expression of intercellular cell adhesion molecule-1 (ICAM-1). SE reduced the increased phosphorylation of nuclear factor kappa B (NFκB) p65 and its subsequent nuclear translocation in retinas from STZ-induced diabetic mice. Results of Western blot and retinal immunofluorescence staining of ionized calcium-binding adapter molecule 1 (Iba1) demonstrated that SE abrogated the activation of microglia cells in STZ-induced diabetic mice.
SE attenuates the development of DR by inhibiting retinal inflammation and restoring the decreased expression of TJ proteins including claudin-1 and claudin-19.
观察[具体提取物名称]乙醇提取物（SE）对糖尿病视网膜病变（DR）的改善作用及其作用机制。
将C57BL/6J小鼠连续5天腹腔注射链脲佐菌素（STZ，55 mg/kg）诱导糖尿病。糖尿病小鼠在注射STZ后1个月口服给予SE（100、200 mg/kg），持续1个月。采用伊文思蓝渗透试验检测血视网膜屏障（BRB）破坏情况。运用实时聚合酶链反应（RT-PCR）、蛋白质免疫印迹法和免疫荧光染色检测mRNA和蛋白质表达。采用酶联免疫吸附测定（ELISA）检测血清肿瘤坏死因子-α（TNF-α）和白细胞介素（IL）-1β含量。
SE（100、200 mg/kg）可逆转STZ诱导的糖尿病小鼠BRB的破坏。SE可逆转紧密连接（TJ）蛋白视网膜claudin-1和claudin-19表达的降低。SE降低了TNF-α和IL-1β升高的血清含量及视网膜mRNA表达。SE还降低了细胞间细胞黏附分子-1（ICAM-1）升高的视网膜表达。SE降低了STZ诱导的糖尿病小鼠视网膜中核因子κB（NFκB）p65磷酸化增加及其随后的核转位。蛋白质免疫印迹法和离子钙结合衔接分子1（Iba1）视网膜免疫荧光染色结果表明，SE消除了STZ诱导的糖尿病小鼠小胶质细胞的激活。
SE通过抑制视网膜炎症和恢复包括claudin-1和claudin-19在内的TJ蛋白降低的表达来减轻DR的发展。
