美洲大蠊提取物PAS840的乙酰胆碱酯酶活性抑制作用及抗氧化能力观察  

Inhibitory effect on acetylcholinesterase activity and antioxidant capacity of Periplaneta Americana extract PAS840

作　　者：管堂飞 杨鑫 洪灿辉 肖培云 张成桂 何正春 GUAN Tangfei;YANG Xin;HONG Canhui;XIAO Peiyun;ZHANG Chenggui;HE Zhengchun(College of Pharmacy,Dali University,Dali 671000,China)

机构地区：[1]大理大学药学院云南省昆虫生物医药研发重点实验室,云南大理671000

出　　处：《山东医药》2024年第11期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(82060747)。

摘　　要：目的探讨美洲大蠊提取物PAS840的乙酰胆碱酯酶(AChE)抑制活性和抗氧化活性。方法根据预实验确定最佳酶促反应条件得到的数据,AChE浓度为0.425U/mL、底物碘化硫代乙酰胆碱(ATCI)浓度为15mmol/L、反应时间为20min,采用改良的Ellman法进行酶促反应,检测不同浓度(7.8125、15.625、31.25、62.5、125、250μg/mL)PAS840对AChE催化ATCI分解的抑制率,计算其半数抑制浓度(IC_(50)),采用米氏方程相关的酶动力学实验测定PAS840对AChE的抑制类型。取不同浓度的PAS840溶液或维生素C(VC),采用1,1-二苯基-2-三硝基苯肼(DPPH)法检测其对DPPH自由基的清除率,采用总还原能力测定法检测其对铁氰化钾的还原率,采用水杨酸法检测其对羟自由基的清除率,并计算相关的IC_(50)。结果PAS840对AChE催化反应进程具有抑制作用,且随着PAS840作用浓度增加和时间延长,其抑制作用更为明显;随着PAS840作用浓度增加,AChE抑制率先逐渐升高再趋于平缓。PAS840抑制AChE催化ATCI分解的IC_(50)为22.30μg/mL,其抑制类型是混合性抑制。随着浓度的增加,PAS840、Vc对DPPH自由基的清除率均呈先升高后平稳的趋势;500μg/mLPAS840对DPPH自由基的清除率为93.95%,同等浓度Vc的清除率为97.31%;PAS840清除DPPH自由基的IC_(50)为43.21μg/mL。随着浓度的增加,PAS840对铁氰化钾的还原率呈先升高后平稳的趋势,Vc对铁氰化钾的还原率呈比较高的平稳趋势;3.0mg/mLPAS840对铁氰化钾的还原率为93.95%,同等浓度Vc的还原率为82.2%;PAS840还原铁氰化钾的IC_(50)为0.498mg/mL。随着浓度的增加,PAS840对羟自由基的清除率呈比较中等的平稳趋势,Vc对羟自由基的清除率呈逐渐升高趋势;1.8mg/mLPAS840对羟自由基的清除率为45.72%,其后逐渐趋于平缓,同等浓度Vc的清除率为74.06%;3.0mg/mLPAS840对羟自由基的清除率为47.83%,同等浓度Vc的清除率为89.73%;PAS840清除羟自由基的IC_(50)为3.65mg/mL。结论PAS840具有较�Objective To investigate the acetylcholinesterase(AChE)inhibitory activity and antioxidant activity of Periplaneta Americana extract PAS840.Methods Based on the data obtained from the preexperiment,the concentration of AChE was fixed at 0.425 U/mL,the concentration of Acetylthiocholine iodide(ATCI)was 15 mmol/L,the reaction time was 20 min,and the enzymatic reaction was performed by the modified Ellman method to detect the inhibitory rates of PAS840 at different concentrations(7.8125,15.625,31.25,62.5,125,and 250μg/mL)on AChE activity.We calculated its half inhibitory concentration(IC_(50)),and determined the type of inhibition of AChE by PAS840 using enzyme kinetic assay related to Michaelis-Menten equation.Different concentrations of PAS840 solution or Vitamin C(VC)were taken to detect the scavenging rate of 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)radicals by the DPPH method,the reduction rate of potassium ferricyanide by the total reducing capacity assay,and the scavenging rate of hydroxyl radicals by the salicylic acid method.Results PAS840 had an inhibitory effect on the process of AChE-catalyzed reaction,and its inhibitory effect became more significant with the increase of PAS840 concentrations and time;with the increase of PAS840 concentrations,the inhibition rate of AChE gradually increased and then leveled off.The IC_(50) of PAS840 to inhibit the decomposition of ATCI catalyzed by AChE was 22.30μg/mL,and its inhibitory behavior was a mixed type of inhibition.The scavenging rates of DPPH radicals by PAS840 and Vc showed a trend of increasing and then leveling off with increased concentrations;the scavenging rates of DPPH radicals by 500μg/mL PAS840 and Vc were 93.95%and 97.31%at the same concentration;the IC_(50) of DPPH radicals scavenging by PAS840 was 43.21μg/mL.The reduction rate of potassium ferricyanide by PAS840 showed a trend of increasing and then leveling off as the concentration increased,but the reduction rate of potassium ferricyanide by Vc showed a continued rising trend;the reducti

关 键 词：PAS840 美洲大蠊 乙酰胆碱酯酶 神经退行性疾病 氧化应激 

分 类 号：R285[医药卫生—中药学]