温肺降浊方对阿尔茨海默病体外模型Aβ25-35诱导PC12细胞凋亡的影响

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中文摘要：目的:探讨温肺降浊方对阿尔茨海默病(AD)体外模型Aβ25-35诱导PC12细胞凋亡的影响.方法:通过β-神经生长因子(β-NGF)分化PC12细胞,Aβ 25-35干预其成为AD细胞模型,CCK-8法确定Aβ25-35干预的最佳浓度和时间点,设立温肺降浊方含药血清低、中、高剂量组进行干预;Transwell检测细胞迁移活性;流式细胞术确定细胞凋亡周期及凋亡比率;qRT-PCR和Western Blot分别检测各组细胞中Bcl-xl、Bad、p53、Caspase-3 mRNA和蛋白的表达情况.结果:β-NGF分化PC12细胞在第6天后成为海马神经元细胞形态结构,制备AD细胞模型以5 μmol/L终浓度的Aβ 25-35干预48 h最佳.与空白组比较,模型组细胞迁移数量显著减少(P＜0.05),细胞凋亡比率显著增加(P＜0.05),Bad、Caspase-3 mRNA及蛋白表达显著上升(P＜0.05),Bcl-xl、p53 mRNA及蛋白表达显著下降(P＜0.05);与模型组比较,温肺降浊方含药血清中、高剂量组细胞迁移数量增加(P＜0.05),细胞凋亡比率显著下降(P＜0.05),温肺降浊方含药血清高剂量组Bad、Caspase-3 mRNA及蛋白表达显著下降(P＜0.05),Bcl-xl、p53 mRNA及蛋白表达显著上升(P＜0.05).结论:温肺降浊方在抑制A β25-35诱导PC12细胞损伤中发挥重要作用,其机制可能与上调Bcl-xl和p53表达并抑制细胞凋亡有关,从而发挥神经保护效应.

外文标题：Effects of Wenfei Jiangzhuo Formula on apoptosis of PC12 cells induced by Aβ25-35 in an in vitro model of Alzheimer disease

外文摘要：Objective:To explore the effect of Wenfei Jiangzhuo Formula on the apoptosis of PC12 cells induced by A β25-35 in an in vitro model of Alzheimer disease(AD).Methods:PC 12 cells were differentiated by β-nerve growth factor(β-NGF),and A β25-35 intervened to become an AD cell model.CCK-8 method was used to determine the optimal concentration and time point of A β25-35 intervention,and low,medium and high dose groups of Wenfei Jiangzhuo Formula-containing serums were set up to intervene;Transwell assay was performed to detect the migration activity of the cells;Flow cytometry was performed to determine the apoptotic cycle and apoptotic ratio;the mRNA and protein expression of Bcl-xl,Bad,p53 and Caspase-3 were detected by qRT-PCR and Western Blot in the cells of each group,respectively.Results:β-NGF differentiated PC 12 cells became hippocampal neuronal cell morphology and structure after day 6,and the AD cell model was prepared with 5 pmol/L final concentration of A β 25-35 intervention for 48 h optimally.Compared with the blank group,the model group showed a significant decrease in the number of cell migration(P＜0.05),an increase in the cell apoptosis rate(P＜0.05),an increase in the expression of Bad and Caspase-3 mRNA and protein(P＜0.05),and a decrease in the expression of Bcl-xl and p53 mRNA and protein(P＜0.05).Compared with the model group,the medium and high doses of Wenfei Jiangzhuo Formula serum-containing showed an increase in the number of cell migration and a decrease in the cell apoptosis rate(P＜0.05);the expression of Bad,Caspase-3 mRNA and protein decreased,and the expression of Bcl-xl,p53 mRNA and protein were increased in the high-dose group of Wenfei Jiangzhuo Formula serum-containing(P＜0.05).Conclusion:Wenfei Jiangzhuo Formula plays an important role in inhibiting Aβ25-35 induced PC 12 cell injury,and its mechanism may be related to the up-regulation of Bcl-xl and p53 expression and the inhibition of apoptosis,thus exerting neuroprotective effects.

外文关键词：

Wenfei Jiangzhuo FormulaA β25-35PC12 cellsAlzheimer disease(AD)Apoptosis

作者：

张鼎、胡芷涵、宋晨曦、李方存、姜明贺、陈炜、胡跃强

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作者单位：

广西中医药大学研究生院,南宁 530001

上海中医药大学,上海 201203

广西中医药大学第一附属医院,南宁 530023

关键词：

温肺降浊方 Aβ25-35 PC12细胞阿尔茨海默病细胞凋亡

基金：

国家自然科学基金地区科学基金项目广西中医脑病临床研究中心项目国家中医药管理局高水平重点学科广西中医药大学"岐黄工程"高层次人才团队广西中医药重点学科建设项目广西中医药大学博士研究生创新项目

项目编号：

82260904桂科AD20238028ZYYZDXK-2023166202410GZXK-Z-20-13YCBXJ2023010

出版年：

2024

中华中医药杂志

中华中医药学会

中华中医药杂志

CSTPCD北大核心

影响因子：1.135

ISSN：1673-1727

年,卷(期)：2024.39(9)