Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, Hubei Province 430060, China; Key Laboratory of Hubei Province for Digestive System Disease, Wuhan, Hubei Province 430060, China; Central Laboratory of Renmin Hospital, Wuhan, Hubei Province 430060, China.
Department of Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China; Central Laboratory of Renmin Hospital, Wuhan, Hubei Province 430060, China.
Int Immunopharmacol. 2020 Nov;88:106862. doi: 10.1016/j.intimp.2020.106862. Epub 2020 Aug 6.
We investigate the effects of Clostridium butyricum(CB) on gut microbiota and colitis associated colon cancer(CAC) in mice.6-8 weeks old C57BL/6 mice were randomly divided into control, azoxymethane (AOM) + dextran sodium sulphate (DSS) and AOM + DSS + CB groups. Mice in the latter two groups received an intraperitoneal injection of AOM (12.5 mg/kg), followed by three cycles of DSS diluted in water (2.5% w/v). Mice in treatment group received CB (2 × 10 CFU in 200 ul normal saline) by gavage administration three times one week. Microbiota composition was assessed by 16S rRNA high-throughput sequencing. Colon samples were collected to examine severity of colitis and tumorigenesis. Cytokines including TNF-a, IL-6 and Cyclo-oxygenase-2 (COX-2) were detected by RT-qPCR. Expression of Bcl-2, Bax and the state of components of NF-κB signaling pathway were detected by western blot. The results revealed that CB regulated structure of intestinal flora and changed the microbial composition; decreased Firmicutes/ Bacteroidetes ratio in phylum level and increased the relative abundance of probiotics; decreased colitis, decreased incidence and size of colorectal cancer(CRC) and increased apoptosis of tumor cells; decreased cytokines including TNF-a and IL-6; decreased level of COX-2; decreased phosphorylation of NF-κB; decreased level of Bcl-2 and increased expression of Bax. In conclusion, CB could regulate structure and composition of gut microbiota and reduces colitis associated colon cancer in mice, the mechanism may be inhibiting NF-κB pathway and promoting apoptosis.
我们研究了丁酸梭菌(CB)对小鼠肠道微生物群和结肠炎相关结肠癌(CAC)的影响。将 6-8 周龄 C57BL/6 小鼠随机分为对照组、氧化偶氮甲烷(AOM)+葡聚糖硫酸钠(DSS)组和 AOM+DSS+CB 组。后两组小鼠腹腔注射 AOM(12.5mg/kg),随后用含 2.5%w/vDSS 的水进行三个周期的 DSS 处理。治疗组小鼠通过灌胃给予 CB(2×10CFU 在 200ul 生理盐水)三次,每周一次。通过 16S rRNA 高通量测序评估微生物群落组成。收集结肠样本以检查结肠炎和肿瘤发生的严重程度。通过 RT-qPCR 检测细胞因子 TNF-a、IL-6 和环氧化酶-2(COX-2)。通过 Western blot 检测 Bcl-2、Bax 和 NF-κB 信号通路成分的状态。结果表明,CB 调节肠道菌群结构并改变微生物组成;在门水平降低厚壁菌门/拟杆菌门比值,增加益生菌相对丰度;减少结肠炎,减少结直肠癌(CRC)的发生率和大小,并增加肿瘤细胞的凋亡;减少细胞因子 TNF-a 和 IL-6;降低 COX-2 水平;减少 NF-κB 的磷酸化;降低 Bcl-2 水平并增加 Bax 的表达。总之,CB 可调节肠道微生物群的结构和组成,并减少小鼠结肠炎相关结肠癌,其机制可能是抑制 NF-κB 通路和促进细胞凋亡。