基于PI3K/AKT/Gsk3β/p53信号通路探究解毒益智方对Aβ25-35诱导PC12细胞神经毒性保护作用机制  

Exploration of the protective mechanism of Jiedu Yizhi formula against Aβ25-35-induced neurotoxicity of PC12 cells based on PI3K/AKT/Gsk3β/p53 signaling pathway

作　　者：冯丽娜 温泉 王杰[3] 王琦 张馨月[3] 黎明全 王庆伟 FENG Lina;WEN Quan;WANG Jie;WANG Qi;ZHANG Xinyue;LI Mingquan;WANG Qingwei(School of Traditional Chinese Medicine,Changchun University of Chinese Medicine,Changchun 130117,China;Department of Encephalopathy,the Third Affiliated Clinical Hospital of Changchun University of Chinese Medicine,Changchun 130117,China;School of Integrated Chinese and Western Medicine,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学中医学院,长春130117 [2]长春中医药大学附属第三临床医院脑病科,长春130117 [3]长春中医药大学中西医结合学院,长春130117

出　　处：《吉林中医药》2022年第12期1439-1444,共6页Jilin Journal of Chinese Medicine

基　　金：吉林省教育厅“十三五”科学技术项目(JJKH20200895KJ);2019年吉林省卫生健康委科技能力提升计划(2019J058)。

摘　　要：目的基于PI3K/AKT/Gsk3β/p53信号通路探究解毒益智方对β淀粉样蛋白25-35片段(Aβ25-35)诱导肾上腺嗜铬细胞瘤(PC12)细胞神经毒性保护作用机制。方法Aβ25-35诱导PC12细胞损伤作为阿尔茨海默病(AD)细胞模型,以解毒益智方含药血清作为治疗药物。实验分为空白组、模型组、解毒益智方(低剂量组、中剂量组及高剂量组)。用噻唑蓝(MTT)法检测细胞存活率及毒性;蛋白免疫印记法(Western Blot,WB)检测磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)、糖原合成酶激酶-3β(GSK3β)蛋白及p53蛋白的表达。结果40μmol/L Aβ25-35为阿尔茨海默病最佳诱导浓度;10%解毒益智方大鼠含药血清为最佳干预浓度;与空白组相比,模型组PC12细胞存活率下降,细胞密度较小,无明显触角,形态类似圆形,且PI3K、p-Akt蛋白表达下调及GSK3β、p53蛋白表达上调(P<0.05);与模型组相比,解毒益智方低、中、高剂量组细胞存活率增多,细胞悬浮死亡现象好转,以高剂量组效果显著;解毒益智方各剂量组PI3K、p-Akt蛋白表达上调及GSK3β、p53蛋白表达均下调(以高剂量组效果显著(P<0.05)。结论解毒益智方可通过调控PI3K/AKT/Gsk3β/p53信号通路,对Aβ25-35诱导PC12细胞神经毒性起保护作用。Objective To explore the neurotoxic protection mechanism of the Jiedu Yizhi formula(JDYZF)on theβ-amyloid 25-35 fragment(Aβ25-35)induced adrenal pheochromocytoma(PC12)cells based on the PI3K/AKT/Gsk3β/p53 signaling pathway.Methods A PC12 cell damage was induced by Aβ25-35 as a model of Alzheimer's disease(AD),and the serum containing the Jiedu Yizhi formula was used as a therapeutic agent.The experiment was divided into a control group,a model group and the JDYZF groups(low-dose group,medium-dose group,and high-dose group).MTT assay was used to detect the cell survival rate and toxicity.The cell survival rate and toxicity were detected by MTT assay.The expressions of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),glycogen synthase kinase-3β(GSK3β)and p53 protein were detected by Western Blot.Results The optimum induced concentration of AD was 40μmol/L Aβ25-35,and the optimum intervention concentration was the rat serum containing 10%Jiedu Yizhi formula.Compared with the control group,the survival rate of PC12 cells in the model group was decreased,the cell density was lower,and there were no obvious antennae,with their morphology similar to a circle,and the expressions of PI3K and p-Akt protein were down-regulated while the expressions of p-GSK3βand p53 protein were up-regulated(P<0.05).Compared with the model group,the cell survival rates in the JDYZF low-,middle-and high-dose groups were increased,and the cell suspension death was improved,with the eff ect most signifi cant in the high-dose group.The expressions of PI3K and p-Akt protein were up-regulated in the JDYZF groups while the expressions of p-GSK3βand p53 protein were down-regulated,with the effect most significant in the highdose group(P<0.05).Conclusion The Jiedu Yizhi formula can play a protective role against the Aβ25-35 induced PC12 cell neurotoxicity by regulating the PI3K/AKT/Gsk3β/p53 signaling pathway.

关 键 词：解毒益智方 阿尔茨海默病 PC12细胞 β淀粉样蛋白 PI3K/AKT/Gsk3β/p53信号通路 

分 类 号：R285.5[医药卫生—中药学]