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脂多糖干预建立体外星形胶质细胞活化模型的 实验研究
孟宪栋亢君君张海锋
0
()
摘要:
目的 探讨脂多糖(LPS)对星形胶质细胞凋亡和活力的影响,建立活化星形胶质细胞模 型。方法 取刚出生1 d 大鼠(P1)大脑进行细胞培养、传代,获得高纯度星形胶质细胞,接种于培养板, 进行LPS(1 μg/ml)干预,设立对照,继续培养24~72 h。按实验要求,细胞凋亡检测分组:LPS-24 h 组、 对照-24 h 组、LPS-72 h 组、对照-72 h 组,使用TUNNEL 法检测各组星形胶质细胞凋亡,并计算凋亡 率;细胞活力检测分组:LPS-0 h组、LPS-24 h组、LPS-72 h组,使用MTT法测定各组星形胶质细胞活力; LPS 干预分组:LPS 干预组和对照组,星形胶质细胞行GFAP免疫组化染色。结果 LPS-24 h 组星形胶 质细胞凋亡率为(7.00±2.23)%,对照-24 h 组为(3.26±1.22)%,两组比较差异无统计学意义(P > 0.05); LPS-72 h 组星形胶质细胞凋亡率为(36.40±5.32)%、对照-72 h 组为(4.00±1.59)%,两组比较差异有统 计学意义(P< 0.05);LPS-0 h组星形胶质细胞活力为(100.23±5.34)%,LPS-24 h组星形胶质细胞活力为 (91.42±9.88)%,LPS-72 h 组星形胶质细胞活力为(51.21±4.58)%,LPS-72 h 组与其余两组比较差异有 统计学意义(P< 0.05)。LPS 干预24 h,星形胶质细胞增生、肥大,突起增多。结论 LPS(1 μg/ml)干预 24 h,星形胶质细胞没有出现明显细胞凋亡和下降的细胞活力,但是形态上出现活化特征,建立了一种 星形胶质细胞活化模型。
关键词:  脂多糖类  星形胶质细胞  炎症  细胞凋亡  活力
DOI:10.3969/j.issn.1009-6574.2021.01.004
基金项目:甘肃省自然科学基金项目(145RJZA039)
Experimental study on establishment of an in vitro activated astrocyte model by intervention oflipopolysaccharide
Meng Xiandong, Kang Junjun, Zhang Haifeng
()
Abstract:
Objective To explore the effect of lipopolysaccharide( LPS) on apoptosis and viability of astrocytes, and to establish an activated astrocyte model. Methods High purity astrocytes were obtained from the brain of 1-day-old rats( P1). The astrocytes were inoculated on the culture plates and treated with LPS( 1 μg/ml) for 24 to 72 hours. According to the experimental requirements, apoptosis was detected using TUNNEL separately in LPS-24 h, control-24 h, LPS-72 h, control-72 h, and then apoptosis rates were calculated; while viability was detected using MTT separately in LPS-0 h, LPS-24 h, and LPS-72 h. Astrocytes were treated with GFAP immunohistochemistry in LPS treatment group and control group. Results The apoptosis rate was( 7.00±2.23)% in LPS-24 h group and( 3.26±1.22)% in control-24 h group, and there was no significant difference between the two groups( P>0.05); the apoptosis rate was( 36.40±5.32)% in LPS-72 h group and( 4.00±1.59)% in control-72 h group, and the difference was statistically significant( P < 0.05); the viability of astrocyte was( 100.23±5.34)%,( 91.42±9.88)%,( 51.21±4.58)% separately in LPS-0 h group, LPS-24 h group, and LPS-72 h group. There was significant difference between LPS-72 h group and other two groups( P< 0.05). After 24 hours of LPS intervention, astrocytes proliferated, enlarged, and their processes increased. Conclusions Treated with LPS( 1 μg/ml) for 24 hours, astrocytes did not show obvious apoptosis and decreased viability, but showed activation in morphology. A model of activated astrocytes might be established.
Key words:  Lipopolysaccharides  Astrocytes  Inflammation  Apoptosis  Viability

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