胃癌脾胃虚弱证的肠道标志菌群分析

Analysis of Intestinal Marker Flora of Deficiency of Spleen and Stomach in Patients with Gastric Cancer

目的探讨胃癌患者脾胃虚弱证形成的肠道微生态学机制。方法收集25例胃癌脾胃虚弱证患者和32例健康体检者作为对照组,高通量测序分析肠道菌群,电化学发光免疫检测血清20种炎症因子,气相色谱-质谱联用分析血清代谢组学。结果与对照组相比,脾胃虚弱证患者肠道细菌变化显著,而肠道真菌变化较小;胃癌脾胃虚弱证患者血清GM-CSF、IL-12/IL-23P40、IL-17α、IL-6水平显著升高,而IL-4、IL-5、TNF-β水平显著降低,12种代谢分子(棕榈酰胺、N1-乙酰精胺、雌马酚、L-酪氨酸、苏氨酸、甲氧基酪胺、赖氨酸、2.3-丁二醇、脯氨酸甜菜碱、组氨酸、羟基丁酸、神经鞘氨醇)显著上升(P<0.05)。线性判别分析显示,胃癌脾胃虚弱证患者肠道5个细菌属(链球菌属、韦荣球菌属、梭杆菌属、Phocaeicola、Pseudarthrobacter)丰度显著升高(P<0.05)。相关性分析显示,肠道韦荣球菌属丰度与12种血清代谢分子均显著正相关(P<0.05);链球菌属与雌马酚、L-酪氨酸、苏氨酸、甲氧基酪胺显著正相关,而与IL-5、TNF-β显著负相关(P<0.05)。结论肠道链球菌属可能是胃癌脾胃虚弱证患者免疫功能低下的潜在标志细菌。

Objective To explore underlying microbial mechanism in the formation of deficiency of spleen and stomach (DSS) in patients with gastric cancer (GC). Methods Here 25 GC patients with DSS were recruited and 32 healthy volunteers were as the control. The gut microbiota were profiled by high throughput sequencing based on 16 S/18 S rRNA genes. Twenty serum inflammatory cytokines were measured by electrochemical luminescence immunoassay, and serum metabolome was analyzed by gas chromatogram and mass spectra (GC/MS). Results Compared to the control, the DSS patients had significantly altered gut bacteria but not gut fungi. Four up-regulated serum cytokines (GM-CSF, IL-12/IL-23 P40, IL-17α and IL-6) and three down-regulated serum cytokines (IL-4, IL-5 and TNF-β) in DSS patients. All the twelve different metabolites were distinctly increased in the DSS patients, which included Palmiticamide, N1-Acetylspermine, Equol, L-tyrosine, Threonin, 3-Methoxytyramine, L-Lysine, 2,3-Butanediol, Homostachydrine, Histidinal, 2-hydroxy-butanoicaci and Sphinganine (P<0.05). Linear discriminant analysis showed that five bacteria genera (Fusobacterium, Phocaeicola, Pseudarthrobacter, Streptococcus and Veillonella) were enriched in DSS patients’ gut. Correlation analysis presented that Veillonella was positively correlated with 12 serum metabolites (P<0.05). Streptococcus was positively correlated with Equol, L-tyrosine, T hreonin and 3-Methoxytyramine, but negatively correlated with IL-5 and TNF-β (P<0.05). Conclusion Increased gut Streptococcus might be the indicator of immunocompromised GC patients with DSS.