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

摘要:

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。

Abstract:

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.