摘要
目的研究黄芩素对β-淀粉样蛋白(amyloidβ-protein,Aβ)诱导PC12细胞损伤的保护作用,探讨其作用机制。方法采用MTT法检测各组细胞活性,筛选黄芩素安全有效浓度;将PC12细胞按随机数字表法分为空白组、Aβ组、黄芩素组、雌二醇组。接种24 h后,黄芩素组给予1×10^(-6)mol/L黄芩素溶液进行干预,雌二醇组给予1×10^(-5)mol/L雌二醇溶液进行干预。2 h后,除空白组外,其余各组加入1.5×10^(-4)mol/L Aβ进行干预造模。培养24 h后使用MTT法测定各组细胞活性。采用氧化试剂盒检测各组细胞SOD、GSH-PX、LDH含量。RT-PCR法检测各组细胞caspase-3 mRNA水平。Western blotting检测各组细胞p-PI3K、p-AKT、caspase-3蛋白表达。结果与Aβ组比较,黄芩素组、雌二醇组PC12细胞存活率[(96.348±0.571)%、(97.183±0.714)%比(86.922±0.429)%]升高(P<0.01);细胞SOD[(54.31±1.34)U/mgprot、(57.38±2.25)U/mgprot比(36.18±2.24)U/mgprot]、GSH-PX[(4.46±0.23)U/mgprot、(4.72±0.31)U/mgprot比(2.05±0.37)U/mgprot]活性升高(P<0.01),LDH[(85.43±0.92)nmol/ml、(82.46±0.27)nmol/ml比(99.17±0.52)nmol/ml]水平降低(P<0.01);细胞caspase-3[(2.24±0.64)、(2.33±0.75)比(3.46±0.46)]mRNA及p-PI3K[(0.46±0.03)、(0.44±0.06)比(0.66±0.09)]、p-AKT[(0.43±0.05)、(0.41±0.02)比(0.58±0.03)]、caspase-3[(0.61±0.03)、(0.56±0.53)比(0.92±0.07)]蛋白表达降低(P<0.01)。结论黄芩素可通过抑制PI3K/AKT通路蛋白表达,减轻细胞凋亡及氧化反应,减少Aβ对PC12细胞的损伤。
Objective To investigate the protective effect of baicalein on injured PC12 cell induced by Aβand explore its mechanism.Methods The method of MTT was used to detect the cell activity of each group and screened the concentration of baicalein.The PC12 cells were randomly divided into the blank group,the Aβgroup,the baicalin group and the estradiol group.24 hours after inoculation,baicalein group was intervened with 1×10^(-6)mol/L baicalein solution,and estradiol group was intervened with 1×10^(-5)mol/L estradiol solution.Two hours later,except the blank group,the other groups were added with 1.5×10^(-4)mol/L Aβto make the model.MTT assay was used to detect the cell viability of each group after 24 hours of cultivation.Then used oxidation kit to detect the contents of superoxide dismutase(SOD),glutathione peroxidase(GSH-PX)and lactate dehydrogenase(LDH)in each group.And the level of caspase-3 mRNA was detected by Real-time Quantitative PCR(RT-PCR).Then the Western blot method was used to detect the expressions of p-PI3K,p-AKT and caspase-3.Results Compared with the Aβgroup,the PC12 cell viability[(96.348±0.571)%,(97.183±0.714)%vs.(86.922±0.429)%]in the baicalin group and the estradiol group significantly increased(P<0.01).The activities of SOD[(54.31±1.34)U/mgprot,(57.38±2.25)U/mgprot vs.(36.18±2.24)U/mgprot]and GSH-PX[(4.46±0.23)U/mgprot,(4.72±0.31)U/mgprot vs.(2.05±0.37)U/mgprot]significantly increased,and the level of LDH[(85.43±0.92)nmol/ml,(82.46±0.27)nmol/ml vs.(99.17±0.52)nmol/ml]significantly decreased(P<0.01).The expression of caspase-3 mRNA(2.24±0.64,2.33±0.75 vs.3.46±0.46)and p-PI3K(0.46±0.03,0.44±0.06 vs.0.66±0.09),p-AKT(0.43±0.05,0.41±0.02 vs.0.58±0.03),caspase-3(0.61±0.03,0.56±0.53 vs.0.92±0.07)protein significantly decreased(P<0.01).Conclusion Baicalein could slow down cell apoptosis and oxidative reaction,reduce the damage of Aβto PC12 cells by inhibiting the expression of PI3K/AKT pathway.
作者
杨柳
王羑彭
张丽宏
李建民
Yang Liu;Wang Youpeng;Zhang Lihong;Li Jianmin(College of Jiamusi,Heilongjiang University of Chinese Medicine,Jiamusi 154007,China)
出处
《国际中医中药杂志》
2021年第7期663-667,共5页
International Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81274035)
黑龙江中医药大学校科研基金(201820)
黑龙江省大学生创新创业训练计划项目(201910228010)。
