目的 采用孟德爾隨機(jī)化（Mendelian randomization，MR）方法評(píng)估腸道菌群與類風(fēng)濕關(guān)節(jié)炎（rheumatoid arthritis，RA）之間的因果關(guān)系，并預(yù)測(cè)能夠通過調(diào)控腸道菌群治療RA的潛在有效中藥。方法 獲取RA與腸道菌群全基因組關(guān)聯(lián)研究數(shù)據(jù)（genome-wide association study，GWAS）。使用R語言進(jìn)行MR分析，采用逆方差加權(quán)法（inverse variance weighted，IVW）探究腸道微生物群與RA之間的潛在因果關(guān)聯(lián)，通過實(shí)施敏感性分析來評(píng)估分析結(jié)果的穩(wěn)健性和可靠性。對(duì)工具變量對(duì)應(yīng)的鄰近基因進(jìn)行基因本體（gene ontology，GO）和京都基因與基因組百科全書（Kyoto encyclopedia of genes and genomes，KEGG）富集分析，探索可能涉及RA發(fā)病機(jī)制的相關(guān)通路。通過Coremine數(shù)據(jù)庫、TCMSP數(shù)據(jù)庫預(yù)測(cè)對(duì)“腸-關(guān)節(jié)”軸存在潛在治療作用的中藥，統(tǒng)計(jì)性味歸經(jīng)及有效成分。結(jié)果 MR分析發(fā)現(xiàn)腸道菌群多爾氏菌屬Dorea（OR＝0.852，95% CI：0.744～0.975，P＝0.020）；副血鏈球菌Streptococcus parasanguinis（OR＝0.918，95% CI：0.847～0.995，P＝0.037）；韋榮球菌屬Veillonella_unclassified（OR＝0.926，95% CI：0.869～0.987，P＝0.018）；毛螺菌科（Lachnospiraceae）細(xì)菌5_1_63FAA（OR＝1.070，95% CI：1.006～1.138，P＝0.031）與RA患病風(fēng)險(xiǎn)存在顯著因果關(guān)系。主要通過病毒蛋白與細(xì)胞因子受體的相互作用、鈣信號(hào)通路及自然殺傷細(xì)胞介導(dǎo)的細(xì)胞毒性通路干預(yù)RA。根據(jù)臨近基因篩選出95味中藥，四氣主要以寒為主，溫、平次之；五味以苦為主，甘、辛次之；歸經(jīng)以肝經(jīng)為主，肺經(jīng)、脾經(jīng)次之；功效以清熱藥為主，補(bǔ)虛藥、活血化瘀藥次之。出現(xiàn)頻次較高的有效成分為β-谷甾醇（β-sitosterol）、槲皮素（quercetin）、山柰酚（kaempferol）、豆甾醇（stigmasterol）。結(jié)論 通過MR分析發(fā)現(xiàn)多爾氏菌屬、副血鏈球菌及韋榮球菌屬Veillonella_unclassified與RA發(fā)病呈負(fù)相關(guān)，毛螺菌科細(xì)菌5_1_63FAA可增加RA發(fā)病風(fēng)險(xiǎn)，富集分析得到了腸道菌作用于RA的生物過程及信號(hào)通路，升麻、白花蛇舌草、蒼耳子、高良姜等中藥可以通過改善腸道微生物群治療RA，其中β-谷甾醇、槲皮素等有效成分具有治療潛力。這些發(fā)現(xiàn)為從“腸-關(guān)節(jié)”軸視角探索RA的中醫(yī)藥防治提供參考，對(duì)RA的達(dá)標(biāo)治療及藥物研發(fā)提供新思路。

Objective To evaluate the causal relationship between gut microbiota (GM) and rheumatoid arthritis (RA) using Mendelian randomization (MR), and predict potential effective traditional Chinese medicines (TCMs) that can regulate gut microbiota to treat RA. Methods The genome-wide association study (GWAS) between RA and GM were obtained. MR analysis was conducted using R language, the potential causal relationship between RA and GM was explored using inverse variance weighting (IVW) method. Sensitivity analysis was conducted to evaluate the robustness and reliability of the analysis results. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment were performed on the analysis on adjacent genes corresponding to instrumental variables to explore potential pathways involved in the pathogenesis of RA. Coremine database and TCMSP database were used to predict potential therapeutic effects of TCMs, statistical flavor meridian analysis, and active ingredients on the “gut joint” axis.Results MR analysis found Dorea (OR＝0.852, 95% CI: 0.744 to 0.975, P＝0.020); Streptococcus parasanguinis (OR＝0.918, 95% CI: 0.847 to 0.995, P＝0.037); Veillonella_ unclassified (OR＝0.926, 95% CI: 0.869 to 0.987, P＝0.018); Lachnospiraceae bacterium 5_1_63FAA (OR＝1.070, 95% CI: 1.006 to 1.138, P＝0.031) had a significant causal relationship with the risk of RA disease. RA was mainly intervened through the interaction of viral protein interaction with cytokine receptor, calcium signaling pathway and natural killer cell mediated cytotoxicity. A total of 95 TCMs were screened according to the proximate genes, and the four qi were cold, followed by warmth and flatness; the five flavors were mainly bitter, followed by sweetness and pungency; the attributed meridians were liver meridian, followed by lung meridian and spleen meridian; and the efficacies were mainly heat-removing drugs, followed by deficiency-tonifying drugs and drugs to activate blood circulation and remove blood stasis. The active ingredients with high frequently were β-sitosterol, quercetin, kaempferol, and stigmasterol. Conclusion Through MR analysis, it was found that Dorea, Streptococcus parasanguinis, and Veillonella unclassified were protective factors for the onset of RA, while Lachnospiraceae bacteria 5_1_63FAA can increase the risk of RA. Enrichment analysis revealed the biological processes and signaling pathways of gut bacteria acting on RA. TCMs such as Shengma (Cimicifugae Rhizoma), Baihua Sheshecao (Hedyotis diffusa), Cangerzi (Xanthii Fructus), and Gaoliangjiang (Alpiniae Officinarum Rhizoma) can treat RA by improving the GM, with active ingredients such as β-sitosterol and quercetin having therapeutic potential. These findings provide reference for exploring the TCMs prevention and treatment of RA from the perspective of the “gut joint” axis, and provide new ideas for the standardized treatment and drug development of RA.

Q811.4;R285

中醫(yī)藥傳承與創(chuàng)新“百千萬”人才工程（岐黃工程）岐黃學(xué)者（20210602-1）;國家中醫(yī)藥管理局名老中醫(yī)傳承工作室（975022）;國家自然科學(xué)基金青年項(xiàng)目“從腸道微生態(tài)失衡探討寒濕外邪加重類風(fēng)濕關(guān)節(jié)炎的病理機(jī)制”（81704000）;中國科學(xué)技術(shù)協(xié)會(huì)青年人才托舉工程（YESS20210352）