基于宿主CD4+T细胞lncRNAs表达和调控研究HAART联用康爱保生丸治疗HIV/AIDS的作用机制

Kang Ai Bao Sheng Wan is a medical institution preparation developed on the basis of the academic thought and formulation theory of researcher Zhang Zhen, a " TCM( traditional Chinese medicine ) master of China"in Yunnan Province.Kang Ai Bao Sheng Wan has been widely used for 10 years in the pilot project of TCM treatment for AIDS in Yunnan Province, and its clinical efficacy has been recognized and focused. Based on the previous research foundation and literature analysis, firstly, chip technology will be applied for screening and verify lncRNAs with abnormal expression in CD4+T cells in HIV/AIDS.Secondly, a prospective clinical study was carried out to observe the effect of HAART combined with Kang Ai Bao Sheng Wan on the differential expression of lncRNAs in CD4+T cells of HIV/AIDS patients. Then, bioinformatics research is carried out to analyze and predict the biological signal pathways that may be enriched by lncRNAs of CD4+T cells in patients' peripheral blood during treatment. Finally, the signal pathway research was carried out, and the biological signal pathway involved in regulation was verified by MilliplexTM EpiQuant. The results of the study can explain the expression and regulatory of lncRNAs on CD4+T cells of HIV/AIDS during the treatment by HAART&Kang Ai Bao Sheng Wan, which is helpful to discover its mechanism and new targets, and provide basis the for TCM&HAART clinical treatment that application and promotion in AIDS.

康爱保生丸是依据云南省“国医大师”张震研究员的学术思想及组方理论研制的医疗机构制剂，已在云南省中医药治疗艾滋病试点项目中广泛应用10年，临床疗效受到认可和关注。综合前期研究基础及文献分析，本项目首先开展转录组学筛选研究，应用芯片技术对不同类型HIV/AIDS患者外周血CD4+T细胞中表达异常的lncRNAs的进行筛查及其验证；其次开展开展前瞻性临床研究，观察HAART联用康爱保生丸治疗对HIV/AIDS患者外周血CD4+T细胞中差异表达lncRNAs的影响作用；而后开展生物信息学研究，分析预测治疗过程中患者外周血CD4+T细胞lncRNAs可能富集的生物学信号通路；最后开展信号通路研究，应用液相悬浮芯片系统验证参与调控的生物学信号通路。研究结果可阐释HAART联用康爱保生丸在治疗HIV/AIDS过程中对免疫细胞异常表达lncRNAs及其调控机制，发现新靶点，为临床应用提供依据.

本项目对两组前瞻性HIV/AIDS队列（HAART联用康爱保生丸组、纯HAART组）外周CD4+T血单核细胞中lncRNAs进行基因芯片的高通量筛选及表达谱分析，发现两组共有28个差异lncRNAs，其中26个下调，2个上调。差异基因GO功能富集与KEGG通路富集分析显示，GO富集分析前10的结果涉及生物过程主要为整合素介导的信号通路、网络蛋白依赖的内吞作用、受体介导的内吞作用正调控、异生物运输、铁离子响应、细胞成分组织调控、金属离子的响应等。此外共获得18个KEGG通路富集分析，其中富集结果前10的为白细胞跨内皮细胞迁移、细菌侵袭上皮细胞信号通路等。信号通路确证实验表明，白细胞跨内皮细胞迁移通路中的FAK、Gi、CAMs、Vinculin表达上调、细菌侵袭上皮细胞通路中FAK、CAMs、Vinculin表达上调。综上研究结果，本研究初步发现HAART联用康爱保生丸可影响HIV/AIDS患者外周血单核细胞中的 lncRNA 表达谱，推测HAART联用康爱保生丸治疗HIV/AIDS患者的机制与调控 lncRNA 及其信号通路的表达水平有关，为HAART联用康爱保生丸治疗HIV/AIDS患者的后续临床应用提供依据及新思路。