基于16S rRNA技术分析柴胡龙骨牡蛎汤对精神分裂症大鼠肠道菌群多样性的影响

目的:基于16S rRNA技术研究柴胡龙骨牡蛎汤对精神分裂症模型大鼠肠道菌群多样性的影响,并探讨该复方治疗精神分裂症的作用机制。方法:雄性SD大鼠除正常组外,每日按剂量0. 1 mg·kg~(-1)腹腔注射地卓西平马来酸盐进行造模,模型成功后,柴胡龙骨牡蛎汤高、中、低剂量组给予实验大鼠柴胡龙骨牡蛎汤(11. 2,5. 6,2. 8 g·kg~(-1)),利培酮组按0. 4 mg·kg~(-1)灌胃给予利培酮片,正常组与模型组灌胃给予蒸馏水,给药体积均为10 mL·kg~(-1),连续给药14 d,将大鼠麻醉后取大鼠盲肠内容物,以Illumina MiSeq为测序平台,对其16S rRNA V4区的肠道菌群可操作分类单元(OTUs)数量,alpha与beta多样性,丰富度指数与多样性指数以及差异菌门与菌属进行综合性分析与评价。结果:柴胡龙骨牡蛎汤可改善精神分裂模型大鼠OTUs数量及alpha与beta多样性的失调,降低菌群生物丰富度指数与多样性指数,对精神分裂症模型大鼠的5种差异菌门(上调拟杆菌门、酸杆菌门、变形菌门和TM7菌门,下调厚壁菌门)与20种差异菌属的分布结构均产生回调作用。结论:柴胡龙骨牡蛎汤对精神分裂症模型大鼠的异常菌群多样性起到治疗作用,并通过16S rRNA技术揭示了肠道菌群在精神分裂状态下的相关病理机制。

Analysis of Effect of Chaihu Longgu Mulitang on Intestinal Microflora Diversity of Schizophrenic Rats Based on 16S rRNA Technique

Objective:To explore the effect of Chaihu Longgu Mulitang on intestinal microflora diversity of schizophrenic model rats, and further reveal its therapeutic characteristics and mechanisms based on the 16S rRNA technique. Method:Except the normal group, male SD rats were intraperitoneally injected dizocilpine maleate with daily dose of 0.1 mg·kg^-1.After the success of the model, Chaihu Longgu Mulitang high, middle and low dose groups were converted into the human clinical upper limit daily, and the experimental rats were given Chaihu Longgu Mulitang with doses of 11.2, 5.6, 2.8 g·kg^-1, respectively. And the positive drug group was treated with 0.4 mg·kg^-1 of risperidone tablets.The normal group and model group was treated with water.The rats were continuous administrated 14 days with dosing volume of 10 mL·kg^-1, the contents in caecum of rats were taken after anesthesia.Illumina MiSeq was used as the sequencing platform, the number of operational taxonomic units(OTUs), richness and diversity indexes, diversity of alpha and beta, differential phylum and genus of intestinal flora in V4 zone of 16S rRNA were comprehensively analyzed and evaluated. Result:Chaihu Longgu Mulitang could improve the number of OTUs, richness and diversity indexes of intestinal flora, imbalance of alpha and beta diversity of schizophrenic model rats.And this formula had a callback effect on 5 differential phyla of bacteria(Bacteroidetes, Acidobacteria, Proteobacteria, Firmicutes and TM7) and 20 genera of bacteria in schizophrenic model rats. Conclusion:Chaihu Longgu Mulitang plays an therapeutic effect on diversity of abnormal microflora in schizophrenic model rats, and this paper reveals the pathological mechanism of intestinal microflora in the state of schizophrenia by 16S rRNA technique.