Zhang Y Y, Li Z D, Jiang N, Wan P X, Deng C B, Su W R, Zhuo Y H
Zhongshan Ophthalmic Center, Sun Yat-sen University, State Key Laboratory of Ophthalmology, Guangzhou 510060, China.
Zhonghua Yan Ke Za Zhi. 2020 May 11;56(5):376-382. doi: 10.3760/cma.j.cn112142-20200107-00011.
To explore the potential neuroprotection effects and associated mechanism of baicalin in a rodent acute hypertensive glaucoma model and oxygen-glucose deprivation/reperfusion (OGD/R) induced retinal ganglion cell (RGC) injury. Experiment research. A rapid and substantial elevation of intraocular pressure was performed to establish an acute hypertensive glaucoma model, and retinal thickness was assessed at 1, 3, 5, and 7 days. The mice were then randomly divided into three groups: normal control group, hypertension group, and baicalin (50 mg/kg) for hypertension group. The effects of baicalin on the RGCs were evaluated by retrograde transporting of Fluoro-Gold. The mRNA levels of tumor necrosis factor-α, interleukine-1β (IL-1β), and inducible nitric oxide synthase were detected by real-time PCR, and the protein levels were measured by Western blot in the retina tissue of acute hypertensive glaucoma model. Purified primary RGC survival under OGD/R stress was measured by flow cytometry, which was also performed to measure the survival rate of RGCs pretreated by different doses of baicalin (2.5 μmol/L, 5.0 μmol/L, and 10.0 μmol/L). The effects of baicalin on primary RGCs co-cultured with mouse microglia cell line BV2 were evaluated by flow cytometry. The cytokine IL-1β in the culture supernatant was measured by immunochemical analyses. Statistical analysis was performed using analysis of variance. Retinal tissue injuries and RGC loss were observed both and . Retinal thickness was decreased to 87.32%±0.94% at 3 days (6.73, 0.01), 74.86%±2.43% at 5 days (13.40, 0.01), and 63.53%±2.15% at 7 days (19.46, 0.01). Treatment of 50 mg/kg baicalin significantly promoted the RGC survival from 61.32%±5.94% to 89.93%±10.08% (4.84, 0.01). Baicalin alleviated the retinal damages by suppressing the expression of inflammatory cytokines as revealed by Western blot and real-time PCR. the RGC survival under OGD/R stress was increased from 51.53%±1.36% to 69.37%±7.09% and 66.23%±4.25% with 5.0, 10.0 μmol/L baicalin administration (5.50, 4.53; both 0.01). BV2 under OGD/R stress did extra damage to RGCs, and baicalin could reverse the damages and increase the survival from 69.37%±7.09% to 73.00%±5.20% (2.82, 0.048) by reducing the release of IL-1β [(39.97±8.76) pg/ml . (61.33±5.78) pg/ml, 4.19, 0.010]. Baicalin could alleviate retina tissue injury directly and promote the survival of RGCs by downregulating the expression of inflammatory cytokines and protecting RGCs from ischemia reperfusion injury. -.
探讨黄芩苷对啮齿动物急性高血压性青光眼模型及氧糖剥夺/再灌注（OGD/R）诱导的视网膜神经节细胞（RGC）损伤的潜在神经保护作用及相关机制。实验研究。通过快速大幅升高眼压建立急性高血压性青光眼模型，并在第1、3、5和7天评估视网膜厚度。然后将小鼠随机分为三组：正常对照组、高血压组和高血压黄芩苷（50 mg/kg）组。通过Fluoro-Gold逆行转运评估黄芩苷对RGCs的影响。通过实时PCR检测急性高血压性青光眼模型视网膜组织中肿瘤坏死因子-α、白细胞介素-1β（IL-1β）和诱导型一氧化氮合酶的mRNA水平，并用蛋白质印迹法测量蛋白质水平。通过流式细胞术测量纯化的原代RGC在OGD/R应激下的存活率，也用于测量用不同剂量黄芩苷（2.5 μmol/L、5.0 μmol/L和10.0 μmol/L）预处理的RGCs的存活率。通过流式细胞术评估黄芩苷对与小鼠小胶质细胞系BV2共培养的原代RGCs的影响。通过免疫化学分析测量培养上清液中的细胞因子IL-1β。使用方差分析进行统计分析。在……时均观察到视网膜组织损伤和RGC丢失。视网膜厚度在第3天降至87.32%±0.94%（6.73，0.01），第5天降至74.86%±2.43%（13.40，0.01），第7天降至63.53%±2.15%（19.46，0.01）。50 mg/kg黄芩苷治疗显著促进RGC存活率从61.32%±5.94%提高到89.93%±10.08%（4.84，0.01）。蛋白质印迹和实时PCR显示，黄芩苷通过抑制炎性细胞因子的表达减轻视网膜损伤。5.0、10.0 μmol/L黄芩苷给药使OGD/R应激下的RGC存活率从51.53%±1.36%提高到69.37%±7.09%和66.23%±4.25%（5.50，4.53；均为0.01）。OGD/R应激下的BV2对RGCs造成额外损伤，黄芩苷可通过减少IL-1β的释放[（39.97±8.76）pg/ml对（61.33±5.78）pg/ml，4.19，0.010]来逆转损伤并使存活率从69.37%±7.09%提高到73.00%±5.20%（2.82，0.048）。黄芩苷可直接减轻视网膜组织损伤，并通过下调炎性细胞因子的表达和保护RGCs免受缺血再灌注损伤来促进RGCs的存活。-。
