目的: 探讨氟化钠(NaF)对BV2小胶质细胞氧糖剥夺/复糖复氧(oxygen-glucose deprivation/reoxygenation, OGD/R)损伤的影响。方法: 构建BV2细胞OGD/R模型,利用不同浓度NaF进行处理,应用噻唑蓝(MTT)法确定模型最佳干预时机。BV2细胞分为对照组(Control)、OGD/R模型组、0.25 mmol/L NaF组、2 mmol/L NaF组、0.25 mmol/L NaF+OGD/R组、2 mmol/L NaF+OGD/R组。除对照组和NaF组外,其余各组在此基础上均需要氧糖剥夺2 h后迅速复糖复氧6 h进行OGD/R造模。采用MTT法检测细胞存活率,相差倒置显微镜下观察细胞形态,比色法检测培养基上清中乳酸脱氢酶(LDH)含量。结果: NaF暴露后BV2细胞黏附力下降,细胞数量明显减少,呈现“阿米巴状”;随着NaF浓度的增加及处理时间的延长,BV2细胞的存活率下降(P＜0.05),并且NaF浓度越高BV2细胞的存活率越低;与Control组相比,OGD/R组BV2细胞存活率下降(P＜0.05);与OGD/R组相比,0.25 mmol/L NaF+OGD/R组和2 mmol/L NaF+OGD/R组BV2细胞存活率均下降(P＜0.05),LDH释放量均增加(P＜0.05);0.25 mmol/L NaF+OGD/R组和2 mmol/L NaF+OGD/R组分别与0.25 mmol/L NaF组、2 mmol/L NaF组相比,BV2细胞的存活率下降(P＜0.05),LDH释放量增加(P＜0.05)。结论: 氟化钠可加重OGD/R诱导的小胶质细胞损伤。
Objective: To investigate the effect of sodium fluoride (NaF) on oxygen-glucose deprivation/rehydration (oxygen-glucose deprivation and reoxygenation,OGD/R) damage in microglia (BV2). Methods: OGD/R model of BV2 cells was constructed, treated with different concentrations of NaF, and used to determine the optimal timing by thiazole blue (methyl thiazolyl tetrazolium, MTT). BV2 cells were divided into control group (Control), OGD/R model group, 0.25 mmol/L NaF group, 2 mmol/L NaF group, 0.25 mmol/L NaF+OGD/R group, and 2 mmol/L NaF+OGD/R group. OGD/R molding was performed for 6 h after oxygen-glucose deprivation, except for control group and NaF groups. Cell viability was measured by MTT. Cell morphology was observed by an optical microscope. And lactate dehydrogenase (lactate dehydrogenase, LDH) content in the medium supernatant was detected by chromatometry. Results: After NaF exposure, the adhesion of BV2 cells decreased, and the number of cells decreased significantly, and the cells morphology changed. With the increase of NaF concentration and treatment time, cell viability decreased(P＜0.05). And the higher the NaF concentration, the lower the viability of BV2 cells. Compared with the control group, Cell viability was significantly decreased in the OGD/R group (P＜0.05). Compared with the OGD/R group, cell viability of BV2 cells were significantly decreased (P＜0.05), and the release of LDH increased significantly in 0.25 mmol/L NaF+OGD/R group and 2 mmol/L NaF+OGD/R group (P＜0.05). Compared with 0.25 mmol/L NaF group and 2 mmol/L NaF+OGD/R group, cell viability of BV2 cells decreased (P＜0.05) and the release of LDH increased (P＜0.05) in 0.25 mmol/L NaF+OGD/R group and 2 mmol/L NaF+OGD/R group, respectively. Conclusion: Sodium fluoride can aggravate microglia damage induced by oxygen-glucose deprivation/reoxygenation.
[1] 魏建明, 曹湘玉, 叶紫阳, 等. 姜黄素调控KLF2表达对BV2细胞OGD模型中的作用及机制[J]. 医学理论与实践, 2024, 37(9): 1441-1444.
[2] 江珊, 俞晓飞. 基于脑出血后小胶质细胞极化及细胞自噬活动探讨中医药干预发挥的抗损伤-促修复双重作用机制的研究进展[J]. 世界科学技术-中医药现代化, 2020, 22(2): 346-352.
[3] 徐飞飞, 崔恺, 王立有, 等. a-细辛醚通过调控NLRP3通路保护氧糖剥夺/再灌注损伤BV2细胞的研究[J]. 中国药理学通报, 2022, 38(8): 1209-1218.
[4] 王薇薇, 郝琦, 项洁. 小鼠脑缺血后SOCS1动态变化及其对氧糖剥夺/复氧后BV2细胞中炎症反应的影响[J]. 中国神经免疫学和神经病学杂志, 2023, 30(2): 115-122.
[5] Liao J, Wei M, Wang J, et al. Naotaifang formula attenuates OGD/R-induced inflammation and ferroptosis by regulating microglial M1/M2 polarization through BMP6/SMADs signaling pathway[J]. Biomed Pharmacother, 2023, 167: 115465.
[6] Zhang Q, Li T, Shi R, et al. Fluoride promotes the secretion of inflammatory factors in microglia through NLRP3/Caspase-1/GSDMD pathway[J]. Environ Sci Pollut Res Int, 2024, 31(13): 19844-19855.
[7] 王正薇, 高霄, 蔡娜, 等. 氟中毒对神经细胞损伤及自噬的影响[J]. 贵州医科大学学报, 2024, 49(5): 625-635.
[8] Yu Q, Shao D, Zhang R, et al. Effects of drinking water fluorosis on L-type calcium channel of hippocampal neurons in mice[J]. Chemosphere, 2019, 220: 169-175.
[9] Chen R, Zhao LD, Liu H, et al. Fluoride induces neuroinflammation and alters wnt signaling pathway in BV2 microglial cells[J]. Inflammation, 2017, 40(4): 1123-1130.
[10] Zhang C, Yang Y, Gao Y, et al. NaF-induced neurotoxicity via activation of the IL-1β/JNK signaling pathway[J]. Toxicology, 2022, 469: 153132.
[11] Kwon HS, Koh SH. Neuroinflammation in neurodegenerative disorders: the roles of microglia and astrocytes[J]. Transl Neurodegener, 2020, 9(1): 42.
[12] Gao X, Su G, Chai M, et al. Research progress on mechanisms of ischemic stroke: regulatory pathways involving microglia[J]. Neurochem Int, 2024, 172: 105656.
[13] Xu X, Gao W, Li L, et al. Annexin A1 protects against cerebral ischemia-reperfusion injury by modulating microglia/macrophage polarization via FPR2/ALX-dependent AMPK-mTOR pathway[J]. J Neuroinflammation, 2021, 18(1): 119.
[14] 玉苏甫·马合木提, 阿布都热合曼·阿卜拉, 苏日青, 等. 抑制转位蛋白在氧糖剥夺/复糖复氧模型中对BV-2细胞损伤和自噬的影响[J]. 中国药理学通报, 2022, 38(5): 761-766.
[15] 叶世泰, 王树超, 王鑫, 等. 姜黄素对缺血性脑卒中大鼠小胶质细胞增殖的影响[J]. 中国医药, 2021, 16(3): 381-384.
