基于16S rRNA技术分析金乌健骨胶囊对胶原诱导关节炎模型大鼠肠道菌群的影响

目的 基于16S rRNA技术研究金乌健骨胶囊对胶原诱导关节炎模型大鼠肠道菌群的影响,并探讨其作用机制。方法 将24只Wistar大鼠随机分为6组,分别为对照组,模型组,金乌健骨胶囊0.225、0.450、1.350g/kg组及甲氨蝶呤（0.001g/kg）组,每组4只。除对照组外,其余各组大鼠制备胶原诱导关节炎模型后ig相应药物,给药期间观察大鼠一般状态并收集大鼠粪便,连续ig 4周后处死大鼠。苏木精–伊红（HE）染色观察各组大鼠关节滑膜及结肠组织病理变化情况,提取粪便基因组DNA,利用Illumina Miseq测序平台对样本16S rRNA进行测序,对测序结果进行生物信息学分析。结果 与模型组大鼠相比,金乌健骨胶囊1.350g/kg组和甲氨蝶呤组关节炎指数评分显著降低（P<0.05、0.01）。6组样本间Chao、observed species、Shannon、Simpson指数没有统计学差异。肠道菌群组成及结构差异分析显示,与对照组相比,模型组普雷沃菌属Prevotella Shan and Collins和螺旋杆菌属Helicobacter Gest and Favinger的相对丰度均增高（P<0.05）,毛螺菌属_NK4A136_group （Lachnospira Bryant and Small_NK4A136_group）的相对丰度降低（P<0.05）。与模型组相比,金乌健骨胶囊0.225g/kg组Lachnospira Bryant and Small_NK4A136_group的相对丰度显著增高（P<0.01）;金乌健骨胶囊0.450g/kg组Lachnospira Bryant and Small_NK4A136_group的相对丰度增高（P<0.05）,Prevotella Shan and Collins和Helicobacter Gest and Favinger的相对丰度降低（P<0.05）。结论 金乌健骨胶囊可能是通过调节肠道菌群的结构组成而发挥治疗类风湿关节炎的作用。

Effect of Jinwu Jiangu Capsules on intestinal flora of collagen-induced arthritis model rats was analyzed based on 16S rRNA technology

Objective To study the effects of Jinwu Jiangu Capsules on intestinal microflora of collagen-induced arthritis model rats based on 16S rRNA technology, and discuss the mechanism of action. Methods A total of 24 Wistar rats were randomly divided into 6 groups: control group, model group, Jinwu Jiangu Capsules 0.225, 0.450, 1.350 g/kg group, and methotrexate (0.001 g/kg) group, with 4 rats in each group. Except the control group, the other groups were given relevant drugs intragastrically after the preparation of collagen-induced arthritis model. During the administration, the general state of the rats was observed and the feces of the rats were collected, and the rats were sacrificed after continuous ig for 4 weeks. The histopathological changes in joint synovium and colon were observed by HE staining. Genome DNA of fecal specimens were extracted and the 16S rRNA were sequenced by Illumina Miseq platform. Sequencing results were analyzed by bioinformatics methodology. Results Compared with model group, the arthritis index scores of Jinwu Jiangu Capsules 1.350 g/kg group and methotrexate group were significantly decreased (P < 0.05, 0.01). There was no statistical difference in Chao, observed species, Shannon, and Simpson indexes among the 6 groups of samples. The analysis of intestinal flora composition and structure showed that compared with the control group, the relative abundance of Prevotella Shan and Collins and Helicobacter Gest and Favinger in the model group was increased (P < 0.05). The relative abundance of the Lachnospira Bryant and Small_NK4A136_group was decreased (P < 0.05). Compared with model group, the relative abundance of Lachnospira Bryant and Small_NK4A136_group in Jinwu Jiangu Capsules 0.225 g/kg group was significantly increased (P < 0.01). Compared with model group, the relative abundance of Lachnospira Bryant and Small_NK4A136_group in Jinwu Jiangu Capsules 0.450 g/kg group were increased (P < 0.05), while the relative abundance of Prevotella Shan and Collins and Helicobacter Gest and Favinger was decreased (P< 0.05). Conclusions Jinwu Jiangu Capsules may play a role in treatment of rheumatoid arthritis by regulating the structural composition of intestinal flora.