Suppression of Berberine and Probiotics (in vitro and in vivo) on the Growth of Colon Cancer With Modulation of Gut Microbiota and Butyrate Production.
Suppression of Berberine and Probiotics (in vitro and in vivo) on the Growth of Colon Cancer With Modulation of Gut Microbiota and Butyrate Production.
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小檗碱和益生菌（体外和体内）通过调节肠道微生物群和丁酸盐的产生来抑制结肠癌的生长
DOI：
10.3389/fmicb.2022.869931
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发表时间：
2022
影响因子：
5.2
通讯作者：
中科院分区：
文献类型：
--
作者：
背景和目的越来越多的证据表明，肠道微生物组在免疫、炎症、代谢和体内平衡中发挥作用，并且由于其在人类健康和疾病（如癌症）之间的平衡而被认为是至关重要的，从而导致针对肠道微生物群的治疗方法的出现。益生菌就是其中之一。然而，关于益生菌在癌症治疗中的作用仍然存在许多挑战。小檗碱（Berberine，BBR）是中药黄连的天然提取物，广泛用于胃肠道疾病的治疗。近年来的研究发现，BBR具有体内、外抗肿瘤作用，但其确切的作用机制尚不清楚。本研究旨在探讨BBR和益生菌在体外和体内对结肠癌细胞生长的抑制作用，以及对肠道微生物组和丁酸产生的调节影响。方法采用结肠癌细胞株HT 29建立裸鼠移植瘤模型和体外模型。将44只裸鼠和HT 29细胞分为对照组、模型组、模型+ BBR组、模型+益生菌组和模型+BBR+益生菌组。双歧杆菌、乳酸杆菌和肠球菌混合活菌粉（LCBLEP）用作益生菌制剂。LCBLEP在厌氧条件下（活菌数应达到1 × 108 CFU）在液体培养基中培养，收集上清液，称之为益生菌上清液（probiotic supernatural，PS）。模型+ BBR组和模型+益生菌组分别在体内给予BBR和LCBLEP或PS治疗4周或在体外治疗48、72和96小时。测量肿瘤体积或细胞增殖。使用16 S rDNA扩增子对肠道微生物群进行焦磷酸测序。用QPCR和ELISA法检测HT 29细胞HDAC 1 mRNA水平和小鼠血清丁酸钠（SB）水平。结果BBR和CBP对荷瘤小鼠均有不同程度的抑瘤作用（p > 0.05），尤以第14天作用最为明显。LCBLEP对肿瘤生长的抑制作用更明显，尤其在11-21天（p < 0.05）。BBR对体外增殖的抑制作用呈浓度依赖性。75%的益生菌培养上清（PS）对细胞增殖的抑制作用最为显著。LCBLEP的添加显著提高了肠道微生物的丰富度和均匀度。BBR显著增加拟杆菌和变形菌的丰度，减少瘤胃球菌，其次是LCBLEP。LCBLEP降低了疣微菌和阿克曼菌的相对丰度，CBP也提高了拟杆菌的相对水平，但降低了疣微菌和阿克曼菌的水平。BBR和LCBLEP或CBP改善了结肠癌裸鼠中肠道微生物的α和β多样性，并显著影响了肠道微生物的生物标志物和代谢功能。BBR和PS处理后HT 29细胞HDAC 1 mRNA水平降低（p < 0.05），SB处理后小鼠血清中SB浓度显著升高（p < 0.05），SB对HT 29细胞增殖的抑制作用强于Panobinostat和TSA。结论虽然BBR与益生菌联合应用在抑制肿瘤生长方面与单用药物相比没有优势，但BBR在抑制结肠癌生长的过程中，通过介导SB的产生，可作为类似益生菌的肠道微生物群调节剂。
Background and Objective An increasing number of evidence has revealed that the gut microbiome functions in immunity, inflammation, metabolism, and homeostasis and is considered to be crucial due to its balance between human health and diseases such as cancer, leading to the emergence of treatments that target intestinal microbiota. Probiotics are one of them. However, many challenges remain regarding the effects of probiotics in cancer treatment. Berberine (BBR), a natural extract of Rhizoma Coptidis and extensively used in the treatment of gastrointestinal diseases, has been found to have antitumor effects in vivo and in vitro by many recent studies, but its definite mechanisms are still unclear. This study aimed to explore the inhibitory effect of BBR and probiotics on the growth of colon cancer cells in vitro and in vivo, and the regulatory influence on the gut microbiome and butyrate production. Methods Colon cancer cell line HT29 was used to establish a xenograft model of nude mice and an in vitro model. A total of 44 nude mice and HT29 cells were divided into control, model, model + BBR, model + probiotics, and model + combination of BBR with probiotics (CBPs). Live combined Bifidobacterium, Lactobacillus, and Enterococcus powder (LCBLEP) was used as a probiotic preparation. LCBLEP was cultured in the liquid medium under anaerobic conditions (the number of viable bacteria should reach 1 × 108CFU), and the supernatant was collected, and it is called probiotic supernatant (PS). Model + BBR and model + probiotics groups were treated with BBR and LCBLEP or PS for 4 weeks in vivo or 48, 72, and 96 h in vitro, respectively. Tumor volume or cell proliferation was measured. Gut microbiota was pyrosequenced using a 16S rDNA amplicon. HDAC1 mRNA level in HT29 cells and sodium butyrate (SB) expression in the serum of mice was detected by QPCR and ELISA. Results The treatment of BBR and CBP reduced the growth of neoplasms in mice to a different extent (p > 0.05), especially at 14 days. The inhibitory effect of LCBLEP on tumor growth was more significant, especially at 11–21 days (p < 0.05). Inhibition of BBR on in vitro proliferation was concentration-dependent. The suppression of 75% probiotic supernatant (PS) on the proliferation was the most significant. The supplement of LCBLEP significantly increased the richness and evenness of the gut microbe. BBR dramatically increased the abundance of Bacteroidetes and Proteobacteria, with reduced Ruminococcus, followed by the LCBLEP. The LCBLEP reduced the relative abundance of Verrucomicrobia and Akkermansia, and the CBP also promoted the relative level of Bacteroidetes but reduced the level of Verrucomicrobia and Akkermansia. BBR and LCBLEP or CBP improved the alpha and beta diversity and significantly affected the biomarker and metabolic function of the gut microbe in nude mice with colon cancer. The level of HDAC1 mRNA was reduced in HT29 cells treated with BBR or PS (p < 0.05), the mice treated with BBR revealed a significantly increased concentration of SB in serum (p < 0.05), and the inhibitory effect of SB on the proliferation of HT29 cells was stronger than panobinostat and TSA. Conclusion Although the combination of BBR and probiotics has no advantage in inhibiting tumor growth compared with the drug alone, BBR can be used as a regulator of the intestinal microbiome similar to the probiotics by mediating the production of SB during reducing the growth of colon cancer.
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影响因子：
5.6
作者：
Hwang S;Yi HC;Hwang S;Jo M;Rhee KJ
通讯作者：
Rhee KJ
影响因子：
29
作者：
Donohoe DR;Garge N;Zhang X;Sun W;O'Connell TM;Bunger MK;Bultman SJ
通讯作者：
Bultman SJ
影响因子：
5.8
作者：
Kim, Sarah;Choi, Sora;Dutta, Moumita;Asubonteng, Jeffrey O.;Polunas, Marianne;Goedken, Michael;Gonzalez, Frank J.;Cui, Julia Yue;Gyamfi, Maxwell A.
通讯作者：
Gyamfi, Maxwell A.
影响因子：
56.9
作者：
LILLY, DM;STILLWELL, RH
通讯作者：
STILLWELL, RH
影响因子：
10.1
作者：
Fan, Lina;Xu, Chaochao;Wang, Liangjing
通讯作者：
Wang, Liangjing