基于 16S rDNA 测序探讨二藤散结胶囊对肠癌恶病质小鼠肠道菌群的影响

Effects of Erteng Sanjie Capsule on intestinal microbiota in colorectal cancer cachexia mice based on 16S rDNA sequencing

关键词:  肠道菌群  癌症恶病质  二藤散结胶囊  CT26  Balb/C小鼠

Key words:  gut microbiota  cancer cachexia  Erteng Sanjie Capsule  CT26  Balb/C mice

摘要:

[目的] 通过皮下接种CT26肠癌细胞建立肠癌恶病质小鼠模型，探索二藤散结胶囊干预对小鼠去瘤体质量、腓肠肌质量、腓肠肌组织病理形态以及肠道微生物结构变化的影响。[方法] 将40只SPF级雄性Balb/C小鼠按照随机数字法分为正常组、模型组、二藤散结胶囊低剂量组[0.75 g/（kg·d）]、二藤散结胶囊中剂量组[1.5 g/（kg·d）]、二藤散结胶囊高剂量组[3 g/（kg·d）]。除正常组外，其余各组均皮下接种CT26肠癌细胞。灌胃14 d后，称量小鼠体质量，收集各组小鼠粪便后处死。取材肿瘤、肝脏、脾脏、腓肠肌并称质量。部分腓肠肌进行苏木精-伊红（HE）染色，观察小鼠腓肠肌的病理学改变；粪便进行16S rDNA高通量测序。[结果] 二藤散结胶囊可以提高肠癌恶病质小鼠去瘤体质量和腓肠肌质量，改善肠癌恶病质小鼠肌肉萎缩情况，丰富肠道微生物物种多样性，调节肠道微生物构成更趋近于正常组，增加un_f_Clostridiales_vadinBB60_group、副拟杆菌属、瘤胃球菌科UCG-014、乳酸杆菌属、un_o_Mollicutes_RF39的相对丰度。[结论] 二藤散结胶囊改善肠癌恶病质导致的肌肉流失、体质量下降和肠道菌群紊乱。

Abstract:

[Objective] To establish a mouse model of cancer cachexia by subcutaneous inoculation of CT26 colon cancer cells，and to investigate the effects of Erteng Sanjie Capsules on tumor-free body weight，gastrocnemius muscle mass，histopathological morphology of the gastrocnemius muscle，and changes in the gut microbiota structure. [Methods] Forty SPF-grade male Balb/C mice were randomly divided into normal group，model group，low-dose Erteng Sanjie Capsule group [0.75 g/（kg·d）]，medium-dose group [1.5 g/（kg·d）]，and high-dose group [3 g/（kg·d）]. Except for normal group，all other groups were subcutaneously inoculated with CT26 colon cancer cells. After 14 days of gavage，the body weight of the mice was measured，and fecal samples were collected before euthanasia. Tumor，liver，spleen，and gastrocnemius muscle tissues were collected and weighed. Part of the gastrocnemius muscle was subjected to HE staining to observe pathological changes，and fecal samples were analyzed by 16S rDNA high-throughput sequencing. [Results] Erteng Sanjie Capsules increased the tumor-free body weight and gastrocnemius muscle mass in mice with cancer cachexia，ameliorated muscle atrophy，enriched the diversity of gut microbiota，and modulated the microbial composition closer to that of the normal group. The relative abundances of un_f_Clostridiales_vadinBB60_group，Parabacteroides，Ruminococcaceae_UCG-014，Lactobacillus，and un_o_Mollicutes_RF39 were significantly increased.[Conclusion] Erteng Sanjie Capsules can ameliorate muscle loss，weight reduction，and gut microbiota dysbiosis induced by cancer cachexia.