基于16S rDNA初探中药对免疫重建不良HIV/AIDS病人肠道菌群的影响

Preliminary study of the effect of traditional Chinese medicine on gut microbiota of HIV/AIDS with poor immune reconstitution based on 16S rDNA

目的基于16S rDNA高通量测序技术,探索经抗病毒治疗(ART)后免疫重建不良的艾滋病病毒(HIV)感染者/艾滋病(AIDS)病人(简称HIV/AIDS病人)经中药治疗24周后肠道菌群的变化。方法采用前后自身对照的方法,于基线和服用参灵固本免疫颗粒、青蒿琥酯片24周后分别采集一次粪便。从粪便样本提取细菌总脱氧核糖核酸(DNA),经16S靶向扩增、高通量测序、物种注释及丰度分析,将菌群丰度与金域现有的健康肠道菌群数据库进行比对,以此为依据描述样本肠道微生态结构及多样性。结果收集了26例经ART后CD4+T淋巴细胞(简称CD4细胞)计数<300个/μL,且血浆HIV载量<50拷贝/mL>18个月的HIV/AIDS病人的粪便。免疫重建不良病人肠道菌群多样性偏低(2.47±0.90),厚壁菌门/拟杆菌门比值偏低0.41(0.25~0.80)。中药治疗24周后,肠型以肠型2占56.5%,肠型3(26.1%)次之,与基线的以肠型3(61.5%)为主,肠型1(19.2%)、2(19.2%)次之相比有统计学差异(P=0.016);乳酸乳球菌丰度0.00(0.00~1.53E-04)、青春双歧杆菌丰度1.66E-04(0.00~3.60E-03)、龙根菌丰度1.31E-04(0.00~2.29E-03),两歧双歧杆菌丰度0.00(0.00~1.21E-04),与基线相比增加,差异有统计学意义。结论免疫重建不良病人肠道微生物平衡失调,服用中药可能改变肠型,增加肠道相关益生菌的丰度。

Objective Based on 16 S rDNA high-throughput sequencing technology,to explore the changes of gut microbiota in HIV-infected/AIDS patients(HIV/AIDS)with poor immune reconstitution after antiretroviral therapy(ART)for 24 weeks with Chinese medicine treatment.Methods Feces were collected respectively at baseline and the treatment of shenling guben immune granules,artesunate for 24 weeks by self-control method.The total deoxyribonucleic acid(DNA)of bacteria extracted from fecal samples was subjected to 16 S targeted amplification,high-throughput sequencing,species annotation,and abundance analysis,and then the flora abundance was compared with the existing healthy intestinal flora database in Kingmed to describe the intestinal microecological structure and diversity of the samples.Results Feces of 26 HIV/AIDS patients were collected with plasma HIV load<50 copies/mL for more than 18 months and CD4+T lymphocyte(CD4 cells)count<300 cell/μL after ART.The diversity of gut microbiota was downregulated(2.47±0.90)and the ratio of firmicutes to bacteroidetes was low as 0.41(0.25-0.80).After 24 weeks of TCM treatment,intestinal type 2(56.5%)was dominant,followed by intestinal type 3(26.1%),with statistically significant differences(P=0.016)compared with baseline intestinal type 3(61.5%),intestinal type 1(19.2%)and intestinal type 2(19.2%).Lactococcus lactis abundance 0.00(0.00-1.53 E-04),bifidobacterium adolescent abundance 1.66 e-04(0.00,3.60 e-03),bifidobacterium longum abundance was 1.31 E-04(0.00-2.29 E-03)and the bifidobacterium bifidum abundance was 0.00(0.00-1.21 E-04).Compared with the baseline,the abundance increased,and the difference was statistically significant.Conclusion Patients with poor immune reconstitution have intestinal microbial imbalance.Use of Chinese medicine may change the intestinal type and increase the abundance of intestinal-related probiotics.