[關(guān)鍵詞]
[摘要]
目的 基于16S rRNA技術(shù)研究金烏健骨膠囊對膠原誘導(dǎo)關(guān)節(jié)炎模型大鼠腸道菌群的影響,并探討其作用機制。方法 將24只Wistar大鼠隨機分為6組,分別為對照組,模型組,金烏健骨膠囊0.225、0.450、1.350g/kg組及甲氨蝶呤(0.001g/kg)組,每組4只。除對照組外,其余各組大鼠制備膠原誘導(dǎo)關(guān)節(jié)炎模型后ig相應(yīng)藥物,給藥期間觀察大鼠一般狀態(tài)并收集大鼠糞便,連續(xù)ig 4周后處死大鼠。蘇木精–伊紅(HE)染色觀察各組大鼠關(guān)節(jié)滑膜及結(jié)腸組織病理變化情況,提取糞便基因組DNA,利用Illumina Miseq測序平臺對樣本16S rRNA進行測序,對測序結(jié)果進行生物信息學(xué)分析。結(jié)果 與模型組大鼠相比,金烏健骨膠囊1.350g/kg組和甲氨蝶呤組關(guān)節(jié)炎指數(shù)評分顯著降低(P<0.05、0.01)。6組樣本間Chao、observed species、Shannon、Simpson指數(shù)沒有統(tǒng)計學(xué)差異。腸道菌群組成及結(jié)構(gòu)差異分析顯示,與對照組相比,模型組普雷沃菌屬Prevotella Shan and Collins和螺旋桿菌屬Helicobacter Gest and Favinger的相對豐度均增高(P<0.05),毛螺菌屬_NK4A136_group (Lachnospira Bryant and Small_NK4A136_group)的相對豐度降低(P<0.05)。與模型組相比,金烏健骨膠囊0.225g/kg組Lachnospira Bryant and Small_NK4A136_group的相對豐度顯著增高(P<0.01);金烏健骨膠囊0.450g/kg組Lachnospira Bryant and Small_NK4A136_group的相對豐度增高(P<0.05),Prevotella Shan and Collins和Helicobacter Gest and Favinger的相對豐度降低(P<0.05)。結(jié)論 金烏健骨膠囊可能是通過調(diào)節(jié)腸道菌群的結(jié)構(gòu)組成而發(fā)揮治療類風濕關(guān)節(jié)炎的作用。
[Key word]
[Abstract]
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.
[中圖分類號]
R9285.5
[基金項目]
國家自然科學(xué)基金資助項目(82060909,82260894);貴州省科技支撐計劃項目(黔科合支撐[2020]4Y155,黔科合平臺人才[2020]2202);貴州省衛(wèi)生健康委科學(xué)技術(shù)基金項目(gzwkj2021-143);貴州省高層次創(chuàng)新型人才培養(yǎng)計劃—“百”層次人才項目(黔科合平臺人才[2016]5650)