摘要
目的:利用人类免疫缺陷病毒(HIV)假病毒刺激的自然杀伤(NK)细胞为研究对象,探讨中医药益艾康对其活化性受体2族成员D(NKG2D)、凝集素样受体亚家族C成员1(NKG2A)、胞质毒性颗粒穿孔素(Perforin)、细胞因子γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF)分泌的影响。方法:磁珠分选HIV阴性健康者外周血NK细胞,选取最适比例的HIV假病毒刺激48 h后,置5%、10%、15%、20%含药血清组及空白对照组,分别加益艾康含药血清及空白鼠血清。流式细胞术检测其活化性受体NKG2D、抑制性受体NKG2A的膜表达;酶联免疫吸附试验(ELISA)检测细胞因子IFN-γ、TNF-α分泌;实时荧光定量PCR检测其NKG2D、穿孔素mRNA表达。结果:NKCell MicroBead Cocktail微珠标记分离NK细胞,最终得到90%以上CD3-CD56^(+)/CD16^(+)NK细胞。与空白对照组比较,5%、10%含药血清组对活化性受体NKG2D有明显的激活作用,5%含药血清组能明显促进IFN-γ的分泌(P<0.05),5%、10%含药血清组显著上调活化性受体NKG2D mRNA表达(P<0.05);5%、10%、15%含药血清组明显促进穿孔素mRNA表达。结论:益艾康可以提高HIV感染患者固有免疫应答,作用机制可能与增强其活化性受体NKG2D的活性,促进其胞质毒性颗粒穿孔素、细胞因子IFN-γ的分泌有关。
Objective:Using natural killer(NK)cells stimulated with human immunodeficiency virus(HIV)pseudovirus as the research model,this study aimed to investigate the effects of the Chinese medicine Yiaikang on the activation receptor natural killer group 2,member D(NKG2D),lectin-like receptor subfamily C member 1(NKG2A),the cytotoxic granule perforin,and the secretion of cytokines interferon-gamma(IFN-γ)and tumor necrosis factor-alpha(TNF-α).Methods:NK cells were isolated from the peripheral blood of HIV-negative healthy donors using magnetic bead sorting.After stimulation with an optimal proportion of HIV pseudovirus for 48 hours,NK cells were treated with Yiaikang-containing serum at concentrations of 5%,10%,15%,and 20%,or with blank mouse serum as a control.Flow cytometry was used to detect membrane expression of NKG2D and NKG2A.ELISA was employed to measure secretion levels of IFN-γand TNF-α.Real-time quantitative PCR was used to assess mRNA expression of NKG2D and perforin.Results:NK cells isolated with the NK Cell MicroBead Cocktail yielded over 90%CD3-CD56^(+)/CD16^(+)NK cells.Compared with the blank control group,the 5%and 10%Yiaikang-containing serum groups significantly activated the expression of NKG2D.The 5%serum group notably promoted IFN-γsecretion(P<0.05).The 5%and 10%serum groups significantly upregulated NKG2D mRNA expression(P<0.05),while the 5%,10%,and 15%serum groups markedly increased perforin mRNA expression.Conclusion:Yiaikang can enhance the innate immune response in HIV-infected patients.Its mechanism may involve increasing the activity of the activation receptor NKG2D and promoting the secretion of perforin and IFN-γ.
作者
钱洁玉
张清燕
杨瑶瑶
邓博文
李杰
岳静宇
李强
QIAN Jieyu;ZHANG Qingyan;YANG Yaoyao;DENG Bowen;LI Jie;YUE Jingyu;LI Qian(AIDS Testing Laboratory of the First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Henan Provincial Key Laboratory of Traditional Chinese Medicine for the Prevention and Treatment of Viral Disease,Zhengzhou 450000,China)
出处
《世界中医药》
北大核心
2025年第17期3081-3085,共5页
World Chinese Medicine
基金
国家自然科学基金项目(82104559,82004207)
河南省科技攻关项目(242102310314,222102310074)。
