A Study on the Effect of Liriopis tuber water extract on Hydrogen Peroxide-stimulated C6 Astrocyte Cells

K. Park, S. Kang, H. Jung, Yong-Ki Park
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Abstract

Objective : To identify the effects of the water extract of Liriope platyphylla tuber (Liriopis tuber, LT) on the activation of astocytes, we investigated the regulatory effects of LT extract on H2O2-induced oxidative damage in C6 rat astrocytes. Methods : LT extract was extracted with boiling water. C6 cell line were treated with LT extract at 1, 2, and 3 mg/ml or without for 30 min and then stimulated with H2O2 at 5 μm for 24 hr. The cell viability was measured by MTT assay. The expression of glial fibrillary acidic protein (GFAP), signal transducer and activator of transcription 3 (STAT3), phospho-STAT3 (pSTAT3), cyclooxygenase (COX-2), Nuclear factor-κB (NF-κB), superoxide dismutase 2 (SOD2), heme oxygenase-1 (HO-1), catalase, Akt, phospho-Akt (p-Akt) phosphoinositide 3-kinases (PI3K), and protein kinase C alpha (PKCα) proteins were determined by Western blot, respectively. GFAP expression was also observed with immunocytochemistry under a fluorescence microscope. Results : LT extract induced cell proliferation in H2O2-stimulated C6 cells. LT extract significantly inhibited the expression of GFAP, NF-κB and COX-2 and increased the expression of HO-1 and the phosphorylation of STAT3 in H2O2-stimulated C6 cells. LT extract also significantly increased the phosphorylation of Akt and decreased the expression of PKCα in a dose-dependent manner in H2O2-stimulated C6 cells. Conclusions : LT extract can regulate H2O2-induced activation of astrocytes through inhibiting the expression of NF-κB, COX-2 and regulating Akt / HO-1, STAT3 or PKCα signaling pathway.1)
百合水提物对过氧化氢刺激C6星形胶质细胞影响的研究
目的:研究白桦叶(Liriope platyphylla tuber, LT)水提物对C6大鼠星形胶质细胞h2o2氧化损伤的调节作用,探讨其对星形胶质细胞活化的影响。方法:用沸水提取大叶提取物。分别用1、2、3 mg/ml或不加浓度的LT提取物处理C6细胞30 min,然后用5 μm浓度的H2O2刺激C6细胞24 h。MTT法测定细胞活力。Western blot检测各组细胞中胶质纤维酸性蛋白(GFAP)、转录信号传导激活因子3 (STAT3)、磷酸化-STAT3 (pSTAT3)、环氧化酶(COX-2)、核因子-κB (NF-κB)、超氧化物歧化酶2 (SOD2)、血红素加氧酶-1 (HO-1)、过氧化氢酶、Akt、磷酸化-Akt (p-Akt)磷酸肌肽3激酶(PI3K)、蛋白激酶Cα (PKCα)蛋白的表达。荧光显微镜下免疫细胞化学观察GFAP的表达。结果:LT提取物可诱导h2o2刺激的C6细胞增殖。在h2o2刺激的C6细胞中,LT提取物显著抑制GFAP、NF-κB和COX-2的表达,增加HO-1的表达和STAT3的磷酸化。在h2o2刺激的C6细胞中,LT提取物也显著增加Akt的磷酸化,并以剂量依赖性的方式降低PKCα的表达。结论:LT提取物可通过抑制NF-κB、COX-2的表达,调节Akt / HO-1、STAT3、PKCα信号通路调节h2o2诱导的星形胶质细胞活化。
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