异戊酰紫草素对β-淀粉样蛋白诱导PC12细胞损伤的保护作用机制研究  

Protective effect and mechanism of isovalerylshikonin on PC12 cells from injury induced by amyloidβ-protein

作　　者：丁文婷 胡杨[1] 朱小南[1] 汪雪兰[1] DING Wen-ting;HU Yang;ZHU Xiao-nan;WANG Xue-lan(Department of Pharmacology,Zhongshan School of Medicine,Sun-Yet Sen University,Guangzhou 510089,Guangdong Province,China)

机构地区：[1]中山大学中山医学院药理教研室,广东广州510089

出　　处：《中国临床药理学杂志》2021年第13期1676-1679,1683,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81460611;81960713);自然科学基金创新基地和人才计划基金资助项目(18JR3RA197);甘肃省中药质量与标准研究重点实验室开放基金资助项目(ZYZL18-008)。

摘　　要：目的研究异戊酰紫草素对β-淀粉样蛋白(Aβ_(25-35))诱导的大鼠嗜铬细胞瘤细胞(PC12细胞)损伤的作用及可能的机制。方法将PC12细胞随机分组为3组:空白对照组、模型组和实验组。空白对照组不做任何处置,模型组加入Aβ_(25-35)40μmol·L^(-1)作用48 h造模,给药组加入5个浓度(0.1,0.5,1.0,2.5和5.0 mol·L^(-1))异戊酰紫草素预孵育4 h后造模。通过细胞计数试剂盒-8法测定细胞的活力,然后选择浓度1μmol·L^(-1)作为实验组进行以下实验。微板法测定培养基内乙酰胆碱(ACh)分泌量,实时荧光定量-PCR法检测乙酰胆碱酯酶(AChE)基因和胆碱乙酰转移酶(ChAT)基因的表达,酶联免疫吸附法测定AChE和ChAT蛋白表达量;比色法测定AChE和ChAT酶活力。结果空白对照组、模型组和实验组的细胞活力分别为(100.16±1.72)%,(49.00±2.60)%和(93.83±2.22)%;这3组的ACh分泌量分别为(65.33±5.03),(46.66±7.63)和(79.33±5.10)mcg·mL^(-1);这3组的AChE蛋白表达量分别为(3.13±0.95),(6.33±3.98)和(3.03±0.87)nmol·L^(-1);这3组的ChAT蛋白表达量分别为(6.13±0.29),(3.23±0.15),(3.18±0.20)nmol·L^(-1);这3组的AChE酶活性分别为(56.97±3.00),(82.70±3.76)和(61.65±2.53)U·mL^(-1);这3组的ChAT酶活性分别为(63.10±4.90),(40.98±2.04)和(81.05±2.70)U·mL^(-1)。基因结果的趋势与蛋白一致。上述指标:模型组和空白对照组比较,差异均有统计学意义(均P<0.05);实验组和模型组比较,差异均有统计学意义(均P<0.05)。结论异戊酰紫草素对Aβ_(25-35)诱导的PC12细胞损伤具有显著的保护作用,这可能与改善PC12细胞胆碱系统功能有关。Objective To explore the effect of isovalerylshikonin(IVS)on PC12 cell injury induced by amyloidβ-protein25-25(Aβ_(25-35))and its possible mechanism.Methods The PC12 cells were randomly grouped into 3 groups:blank control group,model group and experimental group.The blank control group did not do any treatment,the model group was treated with Aβ_(25-35)40μmol·L^(-1)for 48 h,and the drugs group was pre-incubated 4 h with different concentrations(0.1,0.5,1.0,2.5 and 5.0 mol·L^(-1))of isovalerylshikonin before modeling.The cell viability was determined by the cell counting kit method.According to the results of cell viability determination,the concentration of 1μmol·L^(-1)was selected for the following experiments named as experimental group.The secretion of acetylcholine(ACh)was determined by microplate method.The expression of acetylcholinesterase(AChE)gene and choline acetyltransferase(ChAT)gene was detected by real-time fluorescent quantitative-PCR.The protein expression of AChE and ChAT was determined by enzyme-linked immunoadsorption method.The enzyme activity of AChE and ChAT was determined by colorimetric method.Results The cell viability in blank control group,model group and experimental group were(100.16±1.72)%,(49.00±2.60)%and(93.83±2.22)%,respectively the secretion of ACh in the 3 groups were(65.33±5.03),(46.66±7.63)and(79.33±5.10)mcg·mL^(-1),respectively the AChE protein expression levels in the 3 groups were(3.13±0.95),(6.33±3.98)and(3.03±0.87)nmol·L^(-1),respectively;the ChAT protein expression level in the 3 groups were(6.13±0.29),(3.23±0.15)and(3.18±0.20)nmol·L^(-1),respectively;the AChE enzyme activity in the 3 groups were(56.97±3.00),(82.70±3.76)and(61.65±2.53)U·mL^(-1),respectively;the ChAT enzyme activity in the 3 groups were(63.10±4.90),(40.98±2.04)and(81.05±2.70)U·mL^(-1),respectively.The trend of gene results is consistent with that of protein.Comparison between the model group and the blank control group,the differences in the above indicators are statisticall

关 键 词：阿尔茨海默症 异戊酰紫草素 AΒ25-35 乙酰胆碱 乙酰胆碱酯酶 胆碱乙酰转移酶 

分 类 号：R28[医药卫生—中药学]