摘要
目的探讨对氨基水杨酸钠(PAS-Na)对锰(Mn)致大鼠基底核原代星形胶质细胞损伤的拮抗作用,为深入Mn中毒机制及其防治研究提供科学技术平台。方法取SPF级新生24 h内SD大鼠基底核进行消化、传代培养,用荧光免疫法鉴定星形胶质细胞。给予不同浓度Mn(125、250、500和1000μmol/L)和PAS-Na(5、50、500和5000μmol/L)对星形胶质细胞处理24 h,用MTT法测定细胞存活率,按细胞存活率75%为Mn细胞毒性的筛选标准,细胞存活率100%为PAS-Na无细胞毒性标准,选择染Mn和PAS-Na干预剂量。结果染不同浓度Mn 24 h后,星形胶质细胞出现不同程度的肿胀或皱缩,并随染锰浓度增加而加重。正常培养液(对照)、125、250、500和1000μmol/L Mn组细胞存活率依序为100.0%、91.5%、83.3%、78.2%和55.6%。与对照组比较,250、500和1000μmol/L Mn组细胞存活率均降低(P<0.05)。随着Mn浓度增高,细胞存活率呈剂量-反应性下降(r=0.979)。给予5、50、500和5000μmol/L PAS-Na处理24 h后,细胞形态和存活率无明显改变。故选择500μmol/L Mn作为染Mn剂量,50、150和450μmol/L PAS作为干预剂量。与500μmol/L染Mn组比较,150和450μmol/L PAS-Na干预后,细胞皱缩程度减轻,细胞突起增多。150和450μmol/L PAS-Na干预组细胞存活率(89.7%、93.2%)比染Mn组(75.6%)高(P<0.05)。结论染500μmol/L Mn24 h后,星形胶质细胞出现明显的毒性,PAS-Na对染Mn细胞毒性有明显的干预作用,提示PAS-Na对染Mn大鼠原代星形胶质细胞损伤的拮抗作用模型构建成功。
Objective The present study aims to establish antagonistic model of sodium p-aminosalicylic acid(PAS-Na)on manganese(Mn)-induced injury of primary astrocytes from the basal ganglia of rats,to provide a scientific and technical platform for further study on the mechanism of manganism,as well as its prevention and treatment.Methods Astrocytes were obtained from the basal ganglia of newborn SD rats(SPF)within 24 h after digestion and sub-cultured.The purity of astrocytes was identified by using fluorescence immunoassay.The cells were administrated with different doses of Mn(125,250,500,1000μmol/L,respectively)and PAS-Na(5,50,500,5000μmol/L,respectively)for 24 h.Subsequently,cell viability was determined by MTT.The dose of Mn treatment and intervention dose of PAS-Na were selected according to the screening criteria.The study set 75%cell viability rate of control as the screening criteria of Mn cytotoxic dose,and the 100%cell viability rate of control as the non-cytotoxicity standard of PAS-Na.Results Astrocytes showed different degrees of swelling or shrinkage in cell bodies of Mn-treated groups.The cell viabilities of control,125,250,500,1000μmol/L Mn groups were 100.0%,91.5%,83.3%,78.2%,55.6%,respectively.The cell viabilities of 250,500 and 1000μmol/L groups were lower than those in the controls with a dose-dependent manner(r=0.979,P<0.05).While PAS-Na treatment alone has no effects on the cell morphology and viability of astrocytes.Therefore,500μmol/L was selected as the dose of Mn-treatment,and the 50,150,450μmol/L was selected as the intervention dose of PAS-Na.Compared with the 500μmol/L Mn group,the degree of cell shrinkage in 150,450μmol/L PAS-Na intervention groups were reduced.Meanwhile,PAS-Na treatment recovered the decrease number of cell protrusion in astrocytes.Furthermore,the cell viabilities of 150 and 450μmol/L PAS-Na intervention group(89.7%,93.2%,respectively)were higher than those in the Mn group(75.6%,P<0.05).Conclusion The astrocytes shows an obvious toxicity after treatment with 500μmol/L Mn for 24 h.PAS-Na has obvious intervention effect against Mn-induced cytotoxicity,suggesting that the antagonism model of PAS-Na on Mn-induced injury of primary basal ganglia astrocytes is successful.
作者
区仕燕
彭东杰
黄晓薇
李少军
姜岳明
OU Shi-yan;PENG Dong-jie;HUANG Xiao-wei;LI Shao-jun;JIANG Yue-ming(Department of Health Toxicology,School of Public Health,Guangxi Medical University,Nanning Guangxi 530021,China;Guangxi Colleges and Universities Key Laboratory of Prevention and Control of Highly Prevalent Diseases,Guangxi Medical University,Nanning Guangxi 530021,China)
出处
《毒理学杂志》
CAS
CSCD
2020年第1期15-20,共6页
Journal of Toxicology
基金
国家自然科学基金(81460505,81773476).
关键词
对氨基水杨酸钠
锰
基底核
大鼠
原代星形胶质细胞
Sodium p-aminosalicylic acid
Manganese
Basal ganglia
Rat
Primary astrocytes
