首页|穿心莲内酯靶向Akt信号干预HIV感染T细胞的效率研究

穿心莲内酯靶向Akt信号干预HIV感染T细胞的效率研究

Exploring the efficiency of andrographolide targeting Akt signal to interfere with HIV-infected T cells

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目的 探讨穿心莲内酯靶向Akt信号干预人类免疫缺陷病毒(HIV)感染T细胞的效率.方法 培养Jurkat T细胞,用WST-1试剂检测穿心莲内酯干预对Jurkat T细胞活性的影响.将Jurkat T细胞分为空白对照组、穿心莲内酯组、LY294002组(PI3K抑制剂)及穿心莲内酯联合LY294002组.各组药物分别干预24 h后,采用实时荧光定量聚合酶链反应(qRT-PCR)检测各组细胞PI3K、Akt mRNA相对表达量,采用Western blotting检测各组细胞p-PI3K/PI3K、p-Akt/Akt蛋白相对表达量.制备HIV假病毒,用假病毒感染各组细胞,萤光素酶系统检测假病毒感染Jurkat T细胞的效率.结果 WST-1试剂检测结果显示,随着穿心莲内酯浓度增加,Jurkat T细胞活力降低(P<0.05).qRT-PCR和Western blotting检测结果显示,穿心莲内酯降低Jurkat T细胞PI3K及其下游Akt mRNA相对表达量及p-PI3K/PI3K、p-Akt/Akt蛋白相对表达量(P<0.05).HIV假病毒感染实验结果显示,穿心莲内酯降低HIV假病毒感染Jurkat T细胞的效率,联合组比单独组的抑制效果更好(P<0.05).结论 穿心莲内酯可能通过PI3K降低其下游Akt的活化,从而减少HIV假病毒在Jurkat T细胞间的传播及进入细胞的机会,降低其感染Jurkat T细胞的概率.
Objective This study aims to investigate the efficiency of puerarin targeting the Akt signaling pathway in intervening with Human Immunodeficiency Virus(HIV)infection in T cells.Methods Jurkat T cells were cultured,and the impact of puerarin intervention on Jurkat T cell activity was assessed using WST-1 reagent.Jurkat T cells were divided into a blank control group,puerarin group,LY294002 group(PI3K inhibitor),and a combined puerarin-LY294002 group.After 24 hours of drug intervention,real-time quantitative polymerase chain reaction(qRT-PCR)was employed to measure the relative expression of PI3K and Akt mRNA in each group,and Western blotting was used to detect the relative expression of p-PI3K/PI3K and p-Akt/Akt proteins.HIV pseudovirus was prepared to infect cells in each group,and the efficiency of pseudovirus infection in Jurkat T cells was assessed using a luciferase system.Results WST-1 assay results revealed a dose-dependent reduction in Jurkat T cell viability with increasing concentrations of puerarin(P<0.05).qRT-PCR and Western blotting results indicated that puerarin decreased the relative expression of PI3K and downstream Akt mRNA,as well as p-PI3K/PI3K and p-Akt/Akt proteins in Jurkat T cells(P<0.05).Results from the HIV pseudovirus infection experiment demonstrated that puerarin reduced the efficiency of HIV pseudovirus infection in Jurkat T cells,with the combined treatment showing superior inhibitory effects compared to the individual treatment(P<0.05).Conclusion Puerarin may reduce the activation of Akt downstream of PI3K,thereby decreasing the opportunity for HIV pseudovirus spread and entry into Jurkat T cells,ultimately lowering the probability of infection.

acquired immunedeficiency syndromeandrographolidehuman immunodeficiency pseudovirusphosphoinositide 3-kinaseprotein kinase b

张雅靖、鲁世金、魏训东、朱其凤、马洁

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广西中医药大学 研究生院, 广西 南宁 530000

广西中医药大学附属瑞康医院中西医结合转化医学中心, 广西 南宁 530000

北京医院生物治疗中心(国家老年医学中心/中国医学科学院老年医学研究院), 北京 100730

艾滋病 穿心莲内酯 人类免疫缺陷假病毒 PI3K Akt

广西科技基地和人才专项项目广西科技基地和人才专项项目广西研究生教育创新计划项目

2021AC180062021AC19393YCXJ2021093

2024

中国现代医学杂志
中南大学,卫生部肝胆肠外科研究中心

中国现代医学杂志

CSTPCD
影响因子:0.927
ISSN:1005-8982
年,卷(期):2024.34(1)
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