益艾康对艾滋病肺脾气虚证患者外周血宿主限制性因子1表达及干扰素水平的影响

Effect of Yiaikang on SAMHD1 expression and interferon level in peripheral blood of HIV patients with lung-spleen Qi deficiency syndrome

目的研究益艾康治疗对艾滋病肺脾气虚证患者外周血中宿主限制性因子1(sterile alpha motif and histidine-aspartic acid domain-containing protein 1,SAMHD1)表达及干扰素水平的影响,为益艾康治疗艾滋病的作用机制研究提供思路。方法参照AIDS西医及肺脾气虚证中医诊断标准,选择2020年9月—2021年5月河南中医药大学第一附属医院22例艾滋病肺脾气虚证患者(简称“肺脾气虚证患者”)为研究对象(肺脾气虚证组),同时以该地区20例健康人群为对照组,肺脾气虚证患者服用益艾康6个月进行干预治疗。流式细胞表面染色检测T淋巴细胞亚群变化,流式细胞内因子染色检测T细胞中SAMHD1蛋白表达水平,荧光定量聚合酶链反应(polymerase chain reaction,PCR)技术检测外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中SAMHD1 mRNA基因表达水平,酶联免疫吸附技术检测血浆中干扰素α(interferon alpha,IFN-α)、IFN-β、IFN-γ及白细胞介素-2(interleukin-2,IL-2)细胞因子含量。分析肺脾气虚证患者外周血中SAMHD1及干扰素变化,并探讨益艾康的干预作用。结果相较健康对照,肺脾气虚证患者CD4^(+)T淋巴计数、CD4/CD8比值降低(Z=-3.526,P<0.001;Z=-0.881,P<0.001)。T淋巴细胞内SAMHD1蛋白表达上升(SAMHD1 MFI/CD4:Z=-3.425、P=0.001;SAMHD1 MFI/CD8:Z=-2.153、P=0.031)。PBMC细胞中SAMHD1基因表达上升(Z=-3.828,P<0.001)。益艾康治疗可提高患者CD4^(+)T细胞计数和CD4/CD8比值(Z=-2.711,P=0.007;Z=-2.062,P=0.039),降低其SAMHD1 mRNA表达(Z=-2.581,P=0.010),降低IFN-α和IFN-γ水平(Z=-1.985,P=0.047;Z=-2.744,P=0.006)。结论肺脾气虚证患者体内CD4^(+)T细胞数量及CD4/CD8比值降低、SAMHD1及干扰素表达上升。益艾康治疗可升高CD4^(+)T细胞数量及CD4/CD8比值,并降低SAMHD1及干扰素表达水平,进而改善艾滋病病毒感染后的免疫失衡。

Objective To explore the effect of Yiaikang on the expression of sterile alpha motif and histidine-aspartic acid domain-containing protein 1(SAMHD1),a host restriction factor,and interferon levels in peripheral blood of HIV-infected patients with lung-spleen Qi deficiency syndrome.This aims to provide insights into the mechanism of YiaiKang in the treatment of AIDS.Methods According to the diagnostic criteria of Western medicine for AIDS and traditional Chinese medicine for lung-spleen Qi deficiency syndrome,22 cases of HIV patients with lung and spleen Qi deficiency syndrome were enrolled in the study from the First Affiliated Hospital of Henan University of TCM between September 2020 and May 2021,and 20 healthy people in the area were taken as control.The patients with lung and spleen Qi deficiency syndrome were treated ethylwith Yiaikang for 6 months.T lymphocyte subpopulation changes and expression level of SAMHD1 protein in T cells were detected by flow cytometry.SAMHD1 mRNA expression in peripheral blood mononuclear cells(PBMCs)was detected by fluorescence quantitative PCR.The levels of IFN-α,IFN-β,IFN-γ,and IL-2 in plasma were detected by enzyme-linked immunosorbent assay(ELISA).The changes of SAMHD1 and interferon in peripheral blood of HIV patients with lung and spleen Qi deficiency syndrome were analyzed to explore the intervention effect of Yiaikang.Results Compared with healthy people,HIV patients with lung and spleen Qi deficiency syndrome exhibited a reduction in the CD4^(+)T lymphocyte count and CD4/CD8 ratio(Z=-3.526,P<0.001;Z=-0.881,P<0.001).There was an increase in the expression of SAMHD1 protein in T lymphocytes(SAMHD1 MFI/CD4:Z=-3.425,P=0.001;SAMHD1 MFI/CD8:Z=-2.153,P=0.031)and an increase in the expression of SAMHD1 gene in PBMCs(Z=-3.828,P<0.001).Yiaikang treatment significantly increased the CD4^(+)T cell count and CD4/CD8 ratio(Z=-2.711,P=0.007;Z=-2.062,P=0.039),decreased the expression of SAMHD1 mRNA(Z=-2.581,P=0.010),and decreased the levels of IFN-αand IFN-γ(Z=-1.985,P=0.047;Z=-2.744,P=0.006).Conclusions Patients with lung-spleen Qi deficiency syndrome have reduced numbers of CD4^(+)T cells and CD4/CD8 ration,with increased expression of SAMHD1 and interferon factors.Yiaikang treatment can increase the number of CD4^(+)T cells and CD4/CD8 ratio and reduce the expression of SAMHD1 and interferon factor,thereby improving the immune imbalance caused by HIV infection.