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岭南山竹子醇提物对LPS致RAW264.7细胞炎性损伤的保护作用研究
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| 篇名： | 岭南山竹子醇提物对LPS致RAW264.7细胞炎性损伤的保护作用研究 |
| TITLE： | Study on Protective Effects of the Ethanol Extract of Garcinia oblongifolia on LPS-induced RAW 264.7 Cell Inflammatory Injury |
| 摘要： | 目的：探讨岭南山竹子70％乙醇提取物（GOEE）对脂多糖（LPS）致RAW264.7细胞炎性损伤的保护作用及其潜在分子机制。方法：取岭南山竹子果皮用70％乙醇回流提取，制得GOEE。采用Folin-Ciocalteau法和紫外-可见分光光度法分别测定GOEE中总多酚和总黄酮的含量。采用MTT法检测不同剂量GOEE的细胞毒性；以LPS诱导小鼠单核巨噬细胞RAW264.7建立炎症模型，分别以地塞米松、N-乙酰基-L-半胱氨酸为阳性对照，采用Griess法和2′，7′-二氯荧光素法检测细胞培养液中一氧化氮（NO）和细胞中活性氧（ROS）的含量；采用酶联免疫吸附测定（ELISA）法测定细胞培养液中肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）和IL-1β水平；采用Westernblotting法测定细胞中p65蛋白（p65）、磷酸化p65（p-p65）、核因子κB抑制蛋白α（IκBα）、磷酸化IκBα（p-IκBα）、血红素加氧酶-1（HO-1）以及细胞核内核因子Ｅ2相关因子2（NRF2）蛋白的表达水平。结果：GOEE中总多酚与总黄酮的含量分别为（20.191±1.264）、（12.571±0.020）mg/g。当剂量≤500μg/mL时，GOEE对RAW264.7细胞存活率无显著影响（P＞0.05）。与对照组比较，LPS模型组NO、ROS含量，TNF-α、IL-6、IL-1β水平，p-p65/p65和p-IκBα/IκBα比值以及NRF2蛋白的表达水平均显著升高（P＜0.05或P＜0.01）。与LPS模型组比较，阳性对照组和GOEE各处理组细胞NO（除GOEE50μg/mL处理组外）、ROS含量，TNF-α、IL-6、IL-1β水平以及p-p65/p65和p-IκBα/IκBα比值均显著降低，HO-1、NRF2蛋白的表达水平均显著升高（P＜0.05或P＜0.01）。结论：GOEE可通过抑制炎症反应、抑制核因子κB通路磷酸化、促进NRF2蛋白入核等途径来减轻LPS诱导的小鼠巨噬细胞炎性损伤。 |
| ABSTRACT： | OBJECTIVE：To investigate the anti-inflammatory activity of 70％ ethanol extracts from Garcinia oblongifolia （GOEE）on LPS-induced RAW 264.7 cells and its potential molecular mechanism. METHODS ：GOEE was obtained after the fresh G. oblongifolia epicarp refluxed with 70％ ethanol. The contents of total phenol and total flavonoids were determined by Folin-Ciocalteau assay and UV spectrophotometer. MTT assay was used to detect the cytotoxicity of different doses of GOEE. The inflammatory model was induced in RAW 264.7 cells by lipopolysa- ccharide （LPS）. Using dexamethasone and N-acetyl-L-cysteine as positive control ，Griess assay and 2′，7′-dichloro- fluorescein assay were used to detect the contents of NO in cell culture medium and ROS in cells. The levels of TNF-α，IL-6，and IL- 1β in cell culture medium were measured by ELISA. The protein expression of p 65，p-p65，IκBα，p-IκBα，HO-1 in cells and NRF 2 in nucleus were determined by using Western blotting assay. RESULTS：The contents of total phenol and flavonoids in GOEE were （20.191±1.264）and（12.571±0.020）mg/g，respectively. At the concentration below 500 μ g/mL， GOEE had no significantly effect on survival rate of RAW 264.7 cells（P＞ 294043）0.05）. Compared with control group ，the contents of NO and ROS，the levels of TNF-α，IL-6 and IL- 1β，ratio of p-p 65 top65，ratio of p-IκBα to IκBα，protein expression of NRF 2 were increased significantly in LPS model group （P＜0.05 or P＜0.01）. Compared with LPS model group ，the contents of NO（except for GOEE 50 μg/mL group）and ROS ，the levels of TNF-α，IL-6 and IL- 1β，ratio of p-p 65 to p 65 and ratio of p-IκBα to IκBα were decreased significantly in GOEE groups and positive control groups ，while protein expression of HO- 1 and NRF 2 were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS：GOEE attenuates LPS-induced macrophages inflammation injury by inhibiting the inflammatory response and the phosphorylation of NF-κB pathway，promoting NRF 2 protein transportation to the nucleus. |
| 期刊： | 2020年第31卷第14期 |
| 作者： | 黄雷，刘美琼，张小曼，苏少锋，邹小琴，钟小斌，冯洁 |
| AUTHORS： | HUANG Lei，LIU Meiqiong ，ZHANG Xiaoman ，SU Shaofeng ，ZOU Xiaoqin ，ZHONG Xiaobin ，FENG Jie |
| 关键字： | 岭南山竹子；果皮；醇提物；抗炎作用；核因子κB；核因子E2相关因子2；小鼠巨噬细胞；RAW264.7 |
| KEYWORDS： | Garcinia oblongifolia ；Epicarp；Ethanol extract ；Anti-inflammatory effect ；NF-κB；NRF2；Mice macrophage ； |
| 阅读数： | 451 次 |
| 本月下载数： | 5 次 |
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