摘要
目的研究丁酸钠对大鼠系膜细胞增殖、凋亡及对磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物西罗莫司靶蛋白(mTOR)信号通路的影响,并分析可能的机制。方法原代培养并分离、纯化大鼠肾小球系膜细胞。将细胞随机分为空白组、对照组[10μg·L^(-1)表皮生长因子(EGF)]、低剂量实验组(10μg·L^(-1)EGF+0.5 mmol·L^(-1)丁酸钠)、中剂量实验组(10μg·L^(-1)EGF+1.0 mmol·L^(-1)丁酸钠),高剂量实验组10(μg·L^(-1)EGF+2.0 mmol·L^(-1)丁酸钠)。用细胞计数法-8(CCK-8)法检测细胞存活率,用碘化丙啶(PI)染色检测细胞周期,用流式细胞术检测细胞凋亡率,用蛋白质印迹法检测PI3K/AKT/mTOR通道相关蛋白的表达。结果细胞活力检测显示:作用24 h,空白组、对照组和低、中、高剂量实验组的光密度值分别为0.36±0.03、0.66±0.03、0.57±0.05、0.47±0.02和0.41±0.01;作用48 h,空白组、对照组、低剂量实验组、中剂量实验组、高剂量实验组的光密度值分别为0.55±0.03、0.83±0.04、0.68±0.03、0.65±0.02和0.60±0.02,表明10μg·L^(-1)EGF明显刺激系膜细胞增殖活化(P<0.05),而不同浓度丁酸钠干预24、48 h后各组系膜细胞增殖显著受抑制(均P<0.05)。PI染色测定细胞周期发现,空白组、对照组和低、中、高剂量实验组的G1期阻滞率分别为(53.37±0.43)%、(46.84±0.67)%、(57.81±0.48)%、(62.77±0.77)%和(67.57±0.71)%,表明丁酸钠能够诱导细胞周期阻滞于G1期,且呈现浓度依赖性(均P<0.01)。空白组、对照组和低、中、高剂量实验组的细胞凋亡率分别为(4.43±0.48)%、(2.45±0.31)%、(6.16±0.33)%、(8.25±0.40)%和(12.12±0.41)%,低、中、高剂量实验组与对照组比较,差异均有统计学意义(均P<0.01)。结论丁酸钠可有效抑制大鼠系膜细胞的增殖并诱导其凋亡,其作用机制可能与抑制PI3K/AKT/mTOR信号通路活化有关。
Objective To investigate the effect of sodium butyrate on apoptosis of rat mesangial cells and the regulation of phosphatidylinositol3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)signal pathway,and analyze the possible mechanism.Methods Primary cultured,isolated and purified rat mesangial cells.The cells were randomly divided into blank group,control group[10μg·L^(-1)epidermal growth factor(EGF)],low dose experimental group(10μg·L^(-1)EGF+0.5 mmol·L^(-1)sodium butyrate),medium dose experimental group(10μg·L^(-1)EGF+1.0 mmol·L^(-1)sodium butyrate)and high dose experimental group (10μg·L^(-1)EGF+2.0mmol·L^(-1)sodium butyrate).Cell survival rate was detected by cell counting kit-8(CCK-8)method,cell cycle was detected by propyl iodide(PI)staining,cell apoptosis rate was detected by flow cytometry,and the expression of PI3K/AKT/mTOR channel-related proteins was detected by Western blot.Results After 24 h of treatment,the optical density of blank group,control group and low,medium,high dose experimental group were 0.36±0.03,0.66±0.03,0.57±0.05,0.47±0.02,0.41±0.01;which at 48 h of treatment were 0.55±0.03,0.83±0.04,0.68±0.03,0.65±0.02,0.60±0.02,respectively.The results showed that 10μg·L^(-1)EGF significantly stimulated the proliferation and activation of mesangial cells(P<0.05),and the proliferation of mesangial cells was significantly inhibited after treatment with different concentrations of sodium butyrate for 24 and 48 h(P<0.05),PI staining showed that the G1 phase blocking rates of blank group,control group and low,medium,high dose experimental groups were(53.37±0.43)%,(46.84±0.67)%,(57.81±0.48)%,(62.77±0.77)%,(67.57±0.71)%,respectively.The results showed that sodium butyrate could induce cell cycle arrest in G1 phase in a concentrationdependent manner(P<0.01).The apoptosis rates of blank group,control group and low,medium,high dose experimental groups were(4.43±0.48)%,(2.45±0.31)%,(6.16±0.33)%,(8.25±0.40)%and(12.12±0.41)%,the differences were statistically
作者
林志民
蔡志根
施珊红
李渊根
林威远
LIN Zhi-min;CAI Zhi-gen;SHI Shan-hong;LI Yuan-gen;LIN Wei-yuan(Department of Nephrology,The Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,Fujian Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第23期3439-3443,共5页
The Chinese Journal of Clinical Pharmacology
关键词
丁酸钠
大鼠系膜细胞
增殖
凋亡
信号通路
sodium butyrate
rat mesangial cells
proliferate
apoptosis
signal pathway
